出品公司: | Addgene |
---|---|
试剂名称: | pCL-Eco |
质粒类型: | 哺乳动物载体;逆病毒包装;单质粒包装系统 |
高拷贝/低拷贝: | 高拷贝 |
克隆方法: | 限制性内切酶,多克隆位点 |
启动子: | CMV |
载体大小: | 12341 bp |
5' 测序引物及序列: | CMV fwd 5’CGCAAATGGGCGGTAGGCGTG 3’ |
3' 测序引物及序列: | -- |
载体标签: | 无 |
载体抗性: | Ampicillin |
筛选标记: | 无 |
克隆菌株: | DH5α等 |
宿主细胞(系): | 包装细胞系如293T |
备注: | 逆病毒包装载体pCL-Eco是单质粒包装系统,快速产生Helper-free、毒力高的逆病毒颗粒。 |
产品目录号: | #12371 |
稳定性: | 瞬表达 |
组成型/诱导型: | 组成型 |
病毒/非病毒: | 非病毒 |
买家导航
pCL-Eco载体质粒基本信息
pCL-Eco质粒图谱载体图谱和pCL-Eco载体序列质粒序列多克隆位点信息
pCL-Eco质粒载体简介
逆病毒包装载体pCL-Eco的使用方法
293T Growth Medium:
450 ml DMEM (JRH Biosciences)
50 ml FBS (JRH Sciences)
5 ml Glutamine (200 mM in saline, JRH Sciences)
2.5 ml Gentamycin (10 mg/ml in water, Sigma)
HC11 Growth Medium:
5 ml L-Glutamine (200 mM in saline, JRH Sciences)
2.5 ml Bovine Insulin (1 mg/ml in 0.01 N HCL, Sigma)
2.5 ml Gentamycin (10 mg/ml in water, Sigma)
0.5 ml EGF (10 μg/ml in water, Gibco)
50 ml FBS (JRH Sciences)
440 ml RPMI (JRH Sciences)
Day 0: Split 293T cells for transfection:
Wash cells off plate with 10 ml Hanks and put into 15 ml Falcon tube
Add 1 ml 10X trypsin and mix by inversion for about 1-2 minutes
Spin at 1000 rpm in clinical centrifuge for 1-2 minutes
Resuspend cells in 10 ml growth medium
Count cells
Plate cells at 1X106 cells per 100 mm dish
Day 2: Transfect 293T cells:
Plan transfection using 2 μg total DNA and 12 μl FuGene in 200 μl total
volume per 100 mm dish
Dilute Fugene: 12 μl per reaction in serum free DMEM for total volume of
200 μl
Mix DNA plasmids in polypropelene tubes (Falcon #2063):
1 μg DNA of interest
1 μg pCL-Eco
Add 200 μl diluted FuGene dropwise to each DNA tube
Incubate RT 15 min
Feed cells with 5 ml fresh growth media and add FuGene/DNA mixture to media
Day 3: Change medium:
• Remove viral medium and CAREFULLY feed with 6 ml growth medium
This step may reduce a cytostatic factor (produced after transfections) that
can inhibit growth and infection of your target cells.
Split HC11 cells for infection
Seed HC11 cells at 300,000 cells per 100 mm dish
If using coverslips, incubate with FBS for at least 1 hour prior to plating
Day 4: Infect target (HC11) cells:
• Collect virus-containing media off 293T cells
• Syringe filter media through .45 μm filter onto target cells
(Evaporation O/N and loss in filter leaves about 5 ml per plate)
• Add 10 μl 5mg/ml polybrene (final conc = 10 μg polybrene/ml media)
(Store polybrene in aliquots to decrease freeze/thaw cycles)
• Wrap plates in parafilm
• Spin cells in clinical centrifuge 10 min at 1800 rpm
• Rotate plate 1/3. Spin 10 min at 1800 rpm
• Rotate plate 1/3. Spin 10 min at 1800 rpm
• Remove viral media and replace with fresh growth media
Day 6+: Harvest HC11 cells:
• Wash cells with HBSS
• Scrape remaining cells, pellet and flash freeze for protein/RNA analysis
Stain cells for beta-gal expression:
Fixing Solution :
1.35 ml 37% Formaldehyde
0.2 ml Gluteraldehyde
1X PBS to 25 mls
Staining Solution:
625μl 40 mg/ml X-gal in DMF
50 μl 1M MgCL2
750 μl 100 mM Potassium Ferricyanide
750 μl 100 mM Potassium Ferrocyanide
1X PBS to 25 mls
[100 mM Potassium Ferrocyanide = 2.11 g/50ml PBS]
[100 mM Potassium Ferricyanide = 1.64 g/50 ml PBS]
Store at room temperature, protect from light.
1. Make -gal solution and warm in 37°C water bath to prevent crystallization
2. Aspirate off media
3. Wash cells 1X PBS
4. Fix cells 5 min at RT in fixing solution
5. Wash 3X PBS
6 Optional: Add PBS to cells and warm plate at 37°C 5 min (reduces
crystallization)
7. Stain cells with staining solution at 37°C for 1-24 hrs
8. Store stained cells in 70% EtOH at 4°C
(EMBO J. 5: pg 3133, 1986)
Note:
HC11 and MEC primary culture cells are very adherent and can withstand
the force of spinning the plates at 1800 rpm. We found that HC11 cells died at
2200 rpm. However, this is cell type-dependent, and spinning speed should be
optimized accordingly.
Expect to see -gal staining at the periphery of the plate. During the spin
infection the media is pushed to the outside. The cells in the center of the plate
will not be exposed to virus and will not be efficiently infected with retrovirus.
pCL-Eco质粒序列载体序列
LOCUS pCL-Eco Sequencing Result 12341 bp DNA SYN DEFINITION pCL-Eco Sequencing Result ACCESSION KEYWORDS SOURCE ORGANISM other sequences; artificial sequences; vectors. FEATURES Location/Qualifiers source 1..12341 /organism="pCL-Eco Sequencing Result" /mol_type="other DNA" rep_origin 267..886 /label="pBR322_origin" misc_feature complement(1160..1179) /label="SV40pro_F_primer" rep_origin complement(1164..1241) /label="SV40_origin" terminator complement(1709..1828) /label="SV40_PA_terminator" misc_feature complement(1723..1742) /label="EBV_rev_primer" misc_feature complement(2253..2300) /label="SV40_3_splice" misc_feature complement(2306..2321) /label="SV40_int" misc_feature 2540..2562 /label="pGEX_3_primer" CDS complement(3033..5030) /label="ORF frame 1" /translation="MARSTLSKPLKNKVNPRGPLIPLILLMLRGVSTASPGSSPHQVY NITWEVTNGDRETVWATSGNHPLWTWWPDLTPDLCMLAHHGPSYWGLEYQSPFSSPPG PPCCSGGSSPGCSRDCEEPLTSLTPRCNTAWNRLKLDQTTHKSNEGFYVCPGPHRPRE SKSCGGPDSFYCAYWGCETTGRAYWKPSSSWDFITVNNNLTSDQAVQVCKDNKWCNPL VIRFTDAGRRVTSWTTGHYWGLRLYVSGQDPGLTFGIRLRYQNLGPRVPIGPNPVLAD QQPLSKPKPVKSPSVTKPPSGTPLSPTQLPPAGTENRLLNLVDGAYQALNLTSPDKTQ ECWLCLVAGPPYYEGVAVLGTYSNHTSAPANCSVASQHKLTLSEVTGQGLCIGAVPKT HQALCNTTQTSSRGSYYLVAPTGTMWACSTGLTPCISTTILNLTTDYCVLVELWPRVT YHSPSYVYGLFERSNRHKREPVSLTLALLLGGLTMGGIAAGIGTGTTALMATQQFQQL QAAVQDDLREVEKSISNLEKSLTSLSEVVLQNRRGLDLLFLKEGGLCAALKEECCFYA DHTGLVRDSMAKLRERLNQRQKLFESTQGWFEGLFNRSPWFTTLISTIMGPLIVLLMI LLFGPCILNRLVQFVKDRISVVQALVLTQQYHQLKPIEYEP*" CDS complement(4970..8242) /label="ORF frame 2" /translation="MGPMGQPLQVLTLNIEDEYRLHETSKEPDVSLGSTWLSDFPQAW AETGGMGLAVRQAPLIIPLKATSTPVSIKQYPMSQEARLGIKPHIQRLLDQGILVPCQ SPWNTPLLPVKKPGTNDYRPVQDLREVNKRVEDIHPTVPNPYNLLSGLPPSHQWYTVL DLKDAFFCLRLHPTSQPLFAFEWRDPEMGISGQLTWTRLPQGFKNSPTLFDEALHRDL ADFRIQHPDLILLQYVDDLLLAATSELDCQQGTRALLQTLGNLGYRASAKKAQICQKQ VKYLGYLLKEGQRWLTEARKETVMGQPTPKTPRQLREFLGTAGFCRLWIPGFAEMAAP LYPLTKTGTLFNWGPDQQKAYQEIKQALLTAPALGLPDLTKPFELFVDEKQGYAKGVL TQKLGPWRRPVAYLSKKLDPVAAGWPPCLRMVAAIAVLTKDAGKLTMGQPLVILAPHA VEALVKQPPDRWLSNARMTHYQALLLDTDRVQFGPVVALNPATLLPLPEEGLQHNCLD ILAEAHGTRPDLTDQPLPDADHTWYTDGSSLLQEGQRKAGAAVTTETEVIWAKALPAG TSAQRAELIALTQALKMAEGKKLNVYTDSRYAFATAHIHGEIYRRRGLLTSEGKEIKN KDEILALLKALFLPKRLSIIHCPGHQKGHSAEARGNRMADQAARKAAITETPDTSTLL IENSSPYTSEHFHYTVTDIKDLTKLGAIYDKTKKYWVYQGKPVMPDQFTFELLDFLHQ LTHLSFSKMKALLERSHSPYYMLNRDRTLKNITETCKACAQVNASKSAVKQGTRVRGH RPGTHWEIDFTEIKPGLYGYKYLLVFIDTFSGWIEAFPTKKETAKVVTKKLLEEIFPR FGMPQVLGTDNGPAFVSKVSQTVADLLGIDWKLHCAYRPQSSGQVERMNRTIKETLTK LTLATGSRDWVLLLPLALYRARNTPGPHGLTPYEILYGAPPPLVNFPDPDMTRVTNSP SLQAHLQALYLVQHEVWRPLAAAYQEQLDRPVVPHPYRVGDTVWVRRHQTKNLEPRWK GPYTVLLTTPTALKVDGIAAWIHAAHVKAADPGGGPSSRLTWRVQRSQNPLKIRLTRE AP*" misc_feature complement(5079..5095) /label="pBMN_5_primer" /translation="MGPMGQPLQVLTLNIEDEYRLHETSKEPDVSLGSTWLSDFPQAW AETGGMGLAVRQAPLIIPLKATSTPVSIKQYPMSQEARLGIKPHIQRLLDQGILVPCQ SPWNTPLLPVKKPGTNDYRPVQDLREVNKRVEDIHPTVPNPYNLLSGLPPSHQWYTVL DLKDAFFCLRLHPTSQPLFAFEWRDPEMGISGQLTWTRLPQGFKNSPTLFDEALHRDL ADFRIQHPDLILLQYVDDLLLAATSELDCQQGTRALLQTLGNLGYRASAKKAQICQKQ VKYLGYLLKEGQRWLTEARKETVMGQPTPKTPRQLREFLGTAGFCRLWIPGFAEMAAP LYPLTKTGTLFNWGPDQQKAYQEIKQALLTAPALGLPDLTKPFELFVDEKQGYAKGVL TQKLGPWRRPVAYLSKKLDPVAAGWPPCLRMVAAIAVLTKDAGKLTMGQPLVILAPHA VEALVKQPPDRWLSNARMTHYQALLLDTDRVQFGPVVALNPATLLPLPEEGLQHNCLD ILAEAHGTRPDLTDQPLPDADHTWYTDGSSLLQEGQRKAGAAVTTETEVIWAKALPAG TSAQRAELIALTQALKMAEGKKLNVYTDSRYAFATAHIHGEIYRRRGLLTSEGKEIKN KDEILALLKALFLPKRLSIIHCPGHQKGHSAEARGNRMADQAARKAAITETPDTSTLL IENSSPYTSEHFHYTVTDIKDLTKLGAIYDKTKKYWVYQGKPVMPDQFTFELLDFLHQ LTHLSFSKMKALLERSHSPYYMLNRDRTLKNITETCKACAQVNASKSAVKQGTRVRGH RPGTHWEIDFTEIKPGLYGYKYLLVFIDTFSGWIEAFPTKKETAKVVTKKLLEEIFPR FGMPQVLGTDNGPAFVSKVSQTVADLLGIDWKLHCAYRPQSSGQVERMNRTIKETLTK LTLATGSRDWVLLLPLALYRARNTPGPHGLTPYEILYGAPPPLVNFPDPDMTRVTNSP SLQAHLQALYLVQHEVWRPLAAAYQEQLDRPVVPHPYRVGDTVWVRRHQTKNLEPRWK GPYTVLLTTPTALKVDGIAAWIHAAHVKAADPGGGPSSRLTWRVQRSQNPLKIRLTRE AP*" CDS complement(8570..10186) /label="ORF frame 2" /translation="MGQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWP TFNVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVH PKPPPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGAKPKPQVLSDSGGPLIDLLTE DPPPYRDPRPPPSDRDGNGGEATPAGEAPDPSPMASRLRGRREPPVADSTTSQAFPLR AGGNGQLQYWPFSSSDLYNWKNNNPSFSEDPGKLTALIESVLITHQPTWDDCQQLLGT LLTGEEKQRVLLEARKAVRGDDGRPTQLPNEVDAAFPLERPDWDYTTQAGRNHLVHYR QLLLAGLQNAGRSPTNLAKVKGITQGPNESPSAFLERLKEAYRRYTPYDPEDPGQETN VSMSFIWQSAPDIGRKLERLEDLKNKTLGDLVREAEKIFNKRETPEEREERIRRETEE KEERRRTEDEQKEKERDRRRHREMSKLLATVVSGQKQDRQGGERRRSQLDRDQCAYCK EKGHWAKDCPKKPRGPRGPRPQTSLLTLDD*" misc_feature complement(9770..10177) /label="gag" /translation="MGQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWP TFNVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVH PKPPPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGAKPKPQVLSDSGGPLIDLLTE DPPPYRDPRPPPSDRDGNGGEATPAGEAPDPSPMASRLRGRREPPVADSTTSQAFPLR AGGNGQLQYWPFSSSDLYNWKNNNPSFSEDPGKLTALIESVLITHQPTWDDCQQLLGT LLTGEEKQRVLLEARKAVRGDDGRPTQLPNEVDAAFPLERPDWDYTTQAGRNHLVHYR QLLLAGLQNAGRSPTNLAKVKGITQGPNESPSAFLERLKEAYRRYTPYDPEDPGQETN VSMSFIWQSAPDIGRKLERLEDLKNKTLGDLVREAEKIFNKRETPEEREERIRRETEE KEERRRTEDEQKEKERDRRRHREMSKLLATVVSGQKQDRQGGERRRSQLDRDQCAYCK EKGHWAKDCPKKPRGPRGPRPQTSLLTLDD*" misc_feature complement(9783..9799) /label="pBABE_5_primer" /translation="MGQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWP TFNVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVH PKPPPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGAKPKPQVLSDSGGPLIDLLTE DPPPYRDPRPPPSDRDGNGGEATPAGEAPDPSPMASRLRGRREPPVADSTTSQAFPLR AGGNGQLQYWPFSSSDLYNWKNNNPSFSEDPGKLTALIESVLITHQPTWDDCQQLLGT LLTGEEKQRVLLEARKAVRGDDGRPTQLPNEVDAAFPLERPDWDYTTQAGRNHLVHYR QLLLAGLQNAGRSPTNLAKVKGITQGPNESPSAFLERLKEAYRRYTPYDPEDPGQETN VSMSFIWQSAPDIGRKLERLEDLKNKTLGDLVREAEKIFNKRETPEEREERIRRETEE KEERRRTEDEQKEKERDRRRHREMSKLLATVVSGQKQDRQGGERRRSQLDRDQCAYCK EKGHWAKDCPKKPRGPRGPRPQTSLLTLDD*" misc_feature complement(9815..9837) /label="MSCV_primer" /translation="MGQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWP TFNVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVH PKPPPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGAKPKPQVLSDSGGPLIDLLTE DPPPYRDPRPPPSDRDGNGGEATPAGEAPDPSPMASRLRGRREPPVADSTTSQAFPLR AGGNGQLQYWPFSSSDLYNWKNNNPSFSEDPGKLTALIESVLITHQPTWDDCQQLLGT LLTGEEKQRVLLEARKAVRGDDGRPTQLPNEVDAAFPLERPDWDYTTQAGRNHLVHYR QLLLAGLQNAGRSPTNLAKVKGITQGPNESPSAFLERLKEAYRRYTPYDPEDPGQETN VSMSFIWQSAPDIGRKLERLEDLKNKTLGDLVREAEKIFNKRETPEEREERIRRETEE KEERRRTEDEQKEKERDRRRHREMSKLLATVVSGQKQDRQGGERRRSQLDRDQCAYCK EKGHWAKDCPKKPRGPRGPRPQTSLLTLDD*" misc_feature complement(9815..9837) /label="pLXSN_5_primer" /translation="MGQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWP TFNVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVH PKPPPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGAKPKPQVLSDSGGPLIDLLTE DPPPYRDPRPPPSDRDGNGGEATPAGEAPDPSPMASRLRGRREPPVADSTTSQAFPLR AGGNGQLQYWPFSSSDLYNWKNNNPSFSEDPGKLTALIESVLITHQPTWDDCQQLLGT LLTGEEKQRVLLEARKAVRGDDGRPTQLPNEVDAAFPLERPDWDYTTQAGRNHLVHYR QLLLAGLQNAGRSPTNLAKVKGITQGPNESPSAFLERLKEAYRRYTPYDPEDPGQETN VSMSFIWQSAPDIGRKLERLEDLKNKTLGDLVREAEKIFNKRETPEEREERIRRETEE KEERRRTEDEQKEKERDRRRHREMSKLLATVVSGQKQDRQGGERRRSQLDRDQCAYCK EKGHWAKDCPKKPRGPRGPRPQTSLLTLDD*" misc_feature complement(10319..11018) /label="5_LTR" /translation="MGQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWP TFNVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVH PKPPPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGAKPKPQVLSDSGGPLIDLLTE DPPPYRDPRPPPSDRDGNGGEATPAGEAPDPSPMASRLRGRREPPVADSTTSQAFPLR AGGNGQLQYWPFSSSDLYNWKNNNPSFSEDPGKLTALIESVLITHQPTWDDCQQLLGT LLTGEEKQRVLLEARKAVRGDDGRPTQLPNEVDAAFPLERPDWDYTTQAGRNHLVHYR QLLLAGLQNAGRSPTNLAKVKGITQGPNESPSAFLERLKEAYRRYTPYDPEDPGQETN VSMSFIWQSAPDIGRKLERLEDLKNKTLGDLVREAEKIFNKRETPEEREERIRRETEE KEERRRTEDEQKEKERDRRRHREMSKLLATVVSGQKQDRQGGERRRSQLDRDQCAYCK EKGHWAKDCPKKPRGPRGPRPQTSLLTLDD*" promoter complement(10497..11077) /label="CMV_immearly_promoter" /translation="MGQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWP TFNVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVH PKPPPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGAKPKPQVLSDSGGPLIDLLTE DPPPYRDPRPPPSDRDGNGGEATPAGEAPDPSPMASRLRGRREPPVADSTTSQAFPLR AGGNGQLQYWPFSSSDLYNWKNNNPSFSEDPGKLTALIESVLITHQPTWDDCQQLLGT LLTGEEKQRVLLEARKAVRGDDGRPTQLPNEVDAAFPLERPDWDYTTQAGRNHLVHYR QLLLAGLQNAGRSPTNLAKVKGITQGPNESPSAFLERLKEAYRRYTPYDPEDPGQETN VSMSFIWQSAPDIGRKLERLEDLKNKTLGDLVREAEKIFNKRETPEEREERIRRETEE KEERRRTEDEQKEKERDRRRHREMSKLLATVVSGQKQDRQGGERRRSQLDRDQCAYCK EKGHWAKDCPKKPRGPRGPRPQTSLLTLDD*" misc_feature complement(10524..10544) /label="CMV_fwd_primer" /translation="MGQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWP TFNVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVH PKPPPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGAKPKPQVLSDSGGPLIDLLTE DPPPYRDPRPPPSDRDGNGGEATPAGEAPDPSPMASRLRGRREPPVADSTTSQAFPLR AGGNGQLQYWPFSSSDLYNWKNNNPSFSEDPGKLTALIESVLITHQPTWDDCQQLLGT LLTGEEKQRVLLEARKAVRGDDGRPTQLPNEVDAAFPLERPDWDYTTQAGRNHLVHYR QLLLAGLQNAGRSPTNLAKVKGITQGPNESPSAFLERLKEAYRRYTPYDPEDPGQETN VSMSFIWQSAPDIGRKLERLEDLKNKTLGDLVREAEKIFNKRETPEEREERIRRETEE KEERRRTEDEQKEKERDRRRHREMSKLLATVVSGQKQDRQGGERRRSQLDRDQCAYCK EKGHWAKDCPKKPRGPRGPRPQTSLLTLDD*" misc_feature complement(10710..10997) /label="CAG_enhancer" /translation="MGQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWP TFNVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVH PKPPPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGAKPKPQVLSDSGGPLIDLLTE DPPPYRDPRPPPSDRDGNGGEATPAGEAPDPSPMASRLRGRREPPVADSTTSQAFPLR AGGNGQLQYWPFSSSDLYNWKNNNPSFSEDPGKLTALIESVLITHQPTWDDCQQLLGT LLTGEEKQRVLLEARKAVRGDDGRPTQLPNEVDAAFPLERPDWDYTTQAGRNHLVHYR QLLLAGLQNAGRSPTNLAKVKGITQGPNESPSAFLERLKEAYRRYTPYDPEDPGQETN VSMSFIWQSAPDIGRKLERLEDLKNKTLGDLVREAEKIFNKRETPEEREERIRRETEE KEERRRTEDEQKEKERDRRRHREMSKLLATVVSGQKQDRQGGERRRSQLDRDQCAYCK EKGHWAKDCPKKPRGPRGPRPQTSLLTLDD*" misc_feature 11342..11364 /label="pGEX_3_primer" /translation="MGQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWP TFNVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVH PKPPPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGAKPKPQVLSDSGGPLIDLLTE DPPPYRDPRPPPSDRDGNGGEATPAGEAPDPSPMASRLRGRREPPVADSTTSQAFPLR AGGNGQLQYWPFSSSDLYNWKNNNPSFSEDPGKLTALIESVLITHQPTWDDCQQLLGT LLTGEEKQRVLLEARKAVRGDDGRPTQLPNEVDAAFPLERPDWDYTTQAGRNHLVHYR QLLLAGLQNAGRSPTNLAKVKGITQGPNESPSAFLERLKEAYRRYTPYDPEDPGQETN VSMSFIWQSAPDIGRKLERLEDLKNKTLGDLVREAEKIFNKRETPEEREERIRRETEE KEERRRTEDEQKEKERDRRRHREMSKLLATVVSGQKQDRQGGERRRSQLDRDQCAYCK EKGHWAKDCPKKPRGPRGPRPQTSLLTLDD*" promoter 11523..11551 /label="AmpR_promoter" /translation="MGQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWP TFNVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVH PKPPPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGAKPKPQVLSDSGGPLIDLLTE DPPPYRDPRPPPSDRDGNGGEATPAGEAPDPSPMASRLRGRREPPVADSTTSQAFPLR AGGNGQLQYWPFSSSDLYNWKNNNPSFSEDPGKLTALIESVLITHQPTWDDCQQLLGT LLTGEEKQRVLLEARKAVRGDDGRPTQLPNEVDAAFPLERPDWDYTTQAGRNHLVHYR QLLLAGLQNAGRSPTNLAKVKGITQGPNESPSAFLERLKEAYRRYTPYDPEDPGQETN VSMSFIWQSAPDIGRKLERLEDLKNKTLGDLVREAEKIFNKRETPEEREERIRRETEE KEERRRTEDEQKEKERDRRRHREMSKLLATVVSGQKQDRQGGERRRSQLDRDQCAYCK EKGHWAKDCPKKPRGPRGPRPQTSLLTLDD*" gene 11593..112 /label="Ampicillin" /gene="Ampicillin" /translation="MGQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWP TFNVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVH PKPPPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGAKPKPQVLSDSGGPLIDLLTE DPPPYRDPRPPPSDRDGNGGEATPAGEAPDPSPMASRLRGRREPPVADSTTSQAFPLR AGGNGQLQYWPFSSSDLYNWKNNNPSFSEDPGKLTALIESVLITHQPTWDDCQQLLGT LLTGEEKQRVLLEARKAVRGDDGRPTQLPNEVDAAFPLERPDWDYTTQAGRNHLVHYR QLLLAGLQNAGRSPTNLAKVKGITQGPNESPSAFLERLKEAYRRYTPYDPEDPGQETN VSMSFIWQSAPDIGRKLERLEDLKNKTLGDLVREAEKIFNKRETPEEREERIRRETEE KEERRRTEDEQKEKERDRRRHREMSKLLATVVSGQKQDRQGGERRRSQLDRDQCAYCK EKGHWAKDCPKKPRGPRGPRPQTSLLTLDD*" CDS 11593..112 /label="ORF frame 1" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW*" ORIGIN 1 TGGTAAGCCC TCCCGTATCG TAGTTATCTA CACGACGGGG AGTCAGGCAA CTATGGATGA 61 ACGAAATAGA CAGATCGCTG AGATAGGTGC CTCACTGATT AAGCATTGGT AACTGTCAGA 121 CCAAGTTTAC TCATATATAC TTTAGATTGA TTTAAAACTT CATTTTTAAT TTAAAAGGAT 181 CTAGGTGAAG ATCCTTTTTG ATAATCTCAT GACCAAAATC CCTTAACGTG AGTTTTCGTT 241 CCACTGAGCG TCAGACCCCG TAGAAAAGAT CAAAGGATCT TCTTGAGATC CTTTTTTTCT 301 GCGCGTAATC TGCTGCTTGC AAACAAAAAA ACCACCGCTA CCAGCGGTGG TTTGTTTGCC 361 GGATCAAGAG CTACCAACTC TTTTTCCGAA GGTAACTGGC TTCAGCAGAG CGCAGATACC 421 AAATACTGTC CTTCTAGTGT AGCCGTAGTT AGGCCACCAC TTCAAGAACT CTGTAGCACC 481 GCCTACATAC CTCGCTCTGC TAATCCTGTT ACCAGTGGCT GCTGCCAGTG GCGATAAGTC 541 GTGTCTTACC GGGTTGGACT CAAGACGATA GTTACCGGAT AAGGCGCAGC GGTCGGGCTG 601 AACGGGGGGT TCGTGCACAC AGCCCAGCTT GGAGCGAACG ACCTACACCG AACTGAGATA 661 CCTACAGCGT GAGCTATGAG AAAGCGCCAC GCTTCCCGAA GGGAGAAAGG CGGACAGGTA 721 TCCGGTAAGC GGCAGGGTCG GAACAGGAGA GCGCACGAGG GAGCTTCCAG GGGGAAACGC 781 CTGGTATCTT TATAGTCCTG TCGGGTTTCG CCACCTCTGA CTTGAGCGTC GATTTTTGTG 841 ATGCTCGTCA GGGGGGCGGA GCCTATGGAA AAACGCCAGC AACGCGGCCT TTTTACGGTT 901 CCTGGCCTTT TGCTGGCCTT TTGCTCACAT GTTCTTTCCT GCGTTATCCC CTGATTCTGT 961 GGATAACCGT ATTACCGCCT TTGAGTGAGC TGATACCGCT CGCCGCAGCC GAACGACCGA 1021 GCGCAGCGAG TCAGTGAGCG AGGAAGCGGA AGAGCGCCCA ATACGCAAAC CGCCTCTCCC 1081 CGCGCGTTGG CCGATTCATT AATGCAGCCT CCAAAAAAGC CTCCTCACTA CTTCTGGAAT 1141 AGCTCAGAGG CCGAGGCGGC CTCGGCCTCT GCATAAATAA AAAAAATTAG TCAGCCATGG 1201 GGCGGAGAAT GGGCGGAACT GGGCGGAGTT AGGGGCGGGA TGGGCGGAGT AAGACCCGCA 1261 CCCGTTCCCA AACTGCTTAT TAAAGGGGGT ATGGAGGTGC TGGACCTTGT GACAGGGCCA 1321 GACAGTGTGA CAGAAATAGA AGCAGTCCTG TTTTGACAAG TTGCCTCTGG AAGCCTCTAC 1381 AATGCCTCTC TTCTTTTTCT CCAGAGTAAG CGGAGGCCAG GGGCCCCCGG CCTCTGCTTA 1441 ATACTAAAAA AAACAGCTGT TGTCATAGTA ATGATTGGGT GGAAACATTC TAGGCCTGGG 1501 TGGAGAGGCT TTTTGCTTCC TCTTGCAAAA CCACACTGCC CTCTGGAGGG CAGTTGCCTA 1561 GCAACTAATT AAAAGAGGAT GTCGCACGGC CAGCTGCGGT CAGTTAGTCA CTTCCTGCTT 1621 AACTGACTTG ACATTTTCTA TTTTAAGAGT CGGGAGGAAA ATTACTGTGT TGGAGGCCCT 1681 CCGCCATCTT CTGAAGCTGA TCCAGACATG ATAAGATACA TTGATGAGTT TGGACAAACC 1741 ACAACTAGAA TGCAGTGAAA AAAATGCTTT ATTTGTGAAA TTTGTGATGC TATTGCTTTA 1801 TTTGTAACCA TTATAAGCTG CAATAAACAA GTTAACAACA ACAATTGCAT TCATTTTATG 1861 TTTCAGGTTC AGGGGGAGGT GTGGGAGGTT TTTTAAAGCA AGTAAAACCT CTACAAATGT 1921 GGTATGGCTG ATTATGATCT CTAGTCAAGG CACTATACAT CAAATATTTT TCCATAATTT 1981 TCTTGTATAG CAGTGCAGCT TTTTCCTTTG TGGTGTAAAT AGCAAAGCAA GCAAGAGTTC 2041 TATTACTAAA CACAGCATGA CTCAAAAAAC TTAGCAATTC TGAAGGAAAG TCCTTGGGGT 2101 CTTCTACCTT TCTCTTCTTT TTTGGAGGAG TAGAATGTTG AGAGTCAGCA GTAGCCTCAT 2161 CATCACTAGA TGGCATTTCT TCTGAGCAAA ACAGGTTTTC CTCATTAAAG GCATTCCACC 2221 ACTGCTCCCA TTCATCAGTT CCATAGGTTG GAATCTAAAA TACACAAACA ATTAGAATCA 2281 GTAGTTTAAC ACATTATACA CTTAAAAATT TTATATTTAC CTTAGAGCTT TAAATCTCTG 2341 TAGGTAGTTT GTCCAATTAT GTCACACCAC AGAAGTAAGG TTCCTTCACA AAGATCCTCT 2401 AGCTACCTAG CTAGATACAC TCCGCTATCG CTACGTGACT GGGTCATGGC TGCGCCCCGA 2461 CACCCGCCAA CACCCGCTGA CGCGCCCTGA CGGGCTTGTC TGCTCCCGGC ATCCGCTTAC 2521 AGACAAGCTG TGACCGTCTC CGGGAGCTGC ATGTGTCAGA GGTTTTCACC GTCATCACCG 2581 AAACGCGCGA GGCAGCCAGC TTACCTCCCG GTGGTGGGTC GGTGGTCCCT GGGCAGGGGT 2641 CTCCCGATCC CGGACGAGCC CCCAAATGAA AGACCCCCGC TGACGGGTAG TCAATCACTC 2701 AGAGGAGACC CTCCCAAGGA ACAGCGAGAC CACAAGTCGG ATGCAACTGC AAGAGGGTTT 2761 ATTGGATACA CGGGTACCCG GGCGACTCAG TCAATCGGAG GACTGGCGCC CCGAGTGAGG 2821 GGTTGTGGGC TCTTTTATTG AGCTCGGGGA GCAGAAGCGC GCGAACAGAA GCGAGAAGCG 2881 AACTGATTGG TTAGTTCAAA TAAGGCACAG GGTCATTTCA GGTCCTTGGG GCACCCTGGA 2941 AACATCTGAT GGTTCTCTAG CTTGCCAAAC CTACAGGTGG GGTCTTTCAT TCCCCCCTTT 3001 TTCTGGAGAC TAAATAAAAT CTTTTATTTT ATCTATGGCT CGTACTCTAT AGGCTTCAGC 3061 TGGTGATATT GTTGAGTCAA AACTAGAGCC TGGACCACTG ATATCCTGTC TTTAACAAAT 3121 TGGACTAATC GATTAAGAAT GCAGGGTCCG AAGAGCAAAA TCATTAGGAG TACAATGAGG 3181 GGTCCCATAA TGGTAGATAT CAAGGTGGTA AACCAAGGGG ATCTGTTAAA CAGTCCCTCA 3241 AACCATCCTT GAGTTGACTC AAACAGTTTC TGTCTCTGAT TAAGCCTCTC TCTCAATTTG 3301 GCCATGCTGT CTCTCACTAG TCCTGTGTGG TCCGCATAGA AGCAACATTC TTCTTTTAGA 3361 GCAGCACACA GCCCTCCTTC TTTTAGAAAT AACAAGTCTA GGCCCCTTCG ATTCTGTAGG 3421 ACAACTTCAG ACAGGGAAGT GAGAGACTTT TCTAGGTTAG AGATTGATTT TTCAACCTCC 3481 CTGAGATCAT CCTGTACTGC GGCTTGGAGC TGCTGGAATT GCTGAGTGGC CATTAGAGCA 3541 GTAGTCCCTG TTCCTATTCC AGCGGCAATT CCCCCCATGG TTAGTCCACC CAATAATAGG 3601 GCCAGGGTTA ACGACACCGG TTCTCTTTTG TGTCGGTTGG ATCTCTCAAA CAGGCCGTAA 3661 ACATAGCTGG GGGAATGATA GGTGACTCTT GGCCAGAGTT CGACAAGAAC ACAATAATCA 3721 GTGGTAAGGT TCAGTATGGT GGTGGAGATG CATGGAGTAA GCCCGGTACT ACAAGCCCAC 3781 ATGGTACCTG TAGGGGCAAC TAGATAATAG GACCCTCGAC TGCTTGTCTG GGTGGTATTA 3841 CATAGGGCCT GATGTGTTTT GGGAACTGCT CCTATGCAGA GTCCCTGTCC GGTCACTTCG 3901 GACAGGGTCA ACTTGTGTTG GGAGGCCACG GAGCAGTTGG CTGGAGCAGA GGTATGGTTG 3961 GAGTAGGTAC CCAGGACGGC AACCCCTTCG TAGTAGGGGG GTCCCGCTAC TAGACACAAC 4021 CAGCACTCTT GGGTTTTGTC AGGACTGGTG AGGTTGAGGG CTTGGTAGGC TCCGTCTACT 4081 AAGTTTAGCA GCCTATTTTC CGTTCCCGCC GGTGGAAGTT GGGTAGGGGA GAGAGGAGTC 4141 CCACTGGGTG GTTTGGTGAC TGAAGGCGAC TTAACAGGTT TGGGCTTGGA GAGTGGCTGT 4201 TGGTCTGCCA GAACGGGGTT TGGCCCTATT GGGACGCGGG GTCCTAGATT TTGGTATCTG 4261 AGTCGGATCC CAAATGTAAG CCCTGGATCT TGTCCGGAGA CATACAAACG TAAGCCCCAG 4321 TAATGTCCTG TGGTCCAGGA AGTAACCCGT CTCCCGGCGT CTGTAAACCG AATAACTAAG 4381 GGGTTGCACC ACTTATTATC TTTGCATACC TGGACAGCCT GGTCAGAGGT GAGATTGTTG 4441 TTTACTGTGA TGAAATCCCA TGATGAGGAG GGCTTCCAGT AAGCTCTACC GGTTGTCTCA 4501 CAGCCCCAAT AGGCACAGTA GAAGGAGTCT GGACCCCCAC ATGACTTGGA TTCTCGGGGG 4561 CGGTGGGGCC CGGGGCAAAC ATAAAATCCC TCATTTGATT TATGAGTTGT CTGGTCTAGC 4621 TTGAGTCTGT TCCAGGCAGT GTTGCACCGA GGGGTGAGGG AGGTTAAAGG TTCTTCGCAG 4681 TCTCTGGAAC AGCCTGGGCT GCTGCCCCCT GAGCAACAAG GGGGCCCCGG GGGAGAAGAA 4741 AAAGGGGATT GATATTCTAG CCCCCAATAA GATGGTCCAT GGTGGGCTAA CATACATAAA 4801 TCTGGGGTAA GGTCAGGCCA CCAGGTCCAC AGAGGGTGGT TGCCAGAAGT TGCCCATACC 4861 GTCTCCCGAT CTCCATTGGT TACCTCCCAG GTGATATTAT AGACTTGATG AGGACTGGAG 4921 CCGGGCGAAG CAGTACTGAC CCCTCTGAGC ATCAGAAGAA TTAAGGGGAT TAGGGGGCCT 4981 CGCGGGTTAA CCTTATTTTT AAGGGGTTTT GAGAGCGTTG AACGCGCCAT GTCAGTCTAG 5041 AGGATGGTCC ACCCCCGGGG TCGGCAGCCT TCACGTGGGC GGCGTGTATC CAAGCTGCGA 5101 TGCCGTCTAC TTTGAGGGCG GTGGGGGTGG TCAGCAGGAC TGTGTAAGGT CCTTTCCAGC 5161 GAGGTTCTAG GTTCTTAGTC TGGTGTCGGC GGACCCACAC TGTGTCGCCG ACTCGGTAAG 5221 GGTGAGGTAC CACCGGTCGG TCCAGTTGTT CTTGGTAGGC TGCCGCCAGA GGTCTCCAGA 5281 CTTCGTGCTG GACTAAGTAG AGAGCCTGTA AGTGAGCTTG GAGAGAGGGG CTGTTAGTAA 5341 CTCTTGTCAT GTCAGGGTCA GGGAAGTTTA CAAGGGGCGG GGGTGCCCCA TATAAGATCT 5401 CATATGGGGT GAGGCCATGG GGGCCCGGCG TGTTGCGGGC TCGGTACAGG GCTAAGGGGA 5461 GTAGGAGCAC CCAGTCTCTA GAGCCAGTTG CAAGCGTTAA TTTAGTTAAA GTCTCCTTGA 5521 TGGTTCTATT CATTCTTTCT ACCTGGCCTG AGCTTTGGGG TCTGTATGCA CAATGTAATT 5581 TCCAATCAAT CCCCAACAGA TCGGCCACTG TCTGACTCAC CTTGGAGACG AAGGCAGGCC 5641 CATTGTCAGT TCCCAATACC TGAGGCATGC CGAACCTGGG GAAGATCTCC TCTAGTAGCT 5701 TCTTGGTTAC GACCTTGGCG GTTTCTTTCT TGGTTGGGAA GGCTTCTATC CAGCCAGAAA 5761 AGGTATCTAT AAAAACTAGA AGATATTTAT AGCCATACAA TCCGGGCTTT ATCTCGGTGA 5821 AATCGATCTC CCAATGAGTG CCGGGCCGAT GCCCGCGGAC CCTAGTTCCC TGTTTAACGG 5881 CAGACTTGCT GGCGTTGACT TGTGCACAAG CTTTGCAGGT CTCAGTGATA TTTTTGAGTG 5941 TTCGATCCCG GTTCAGCATG TAGTAGGGAC TGTGGCTTCT CTCTAGGAGA GCCTTCATTT 6001 TTGAGAAGCT GAGGTGAGTC AGCTGATGAA GAAAGTCTAA TAATTCAAAA GTAAACTGGT 6061 CAGGCATCAC AGGTTTTCCT TGGTAGACCC AATACTTCTT TGTTTTATCA TAAATGGCCC 6121 CCAACTTGGT TAGGTCCTTT ATATCAGTCA CTGTGTAATG AAAATGTTCT GAGGTGTAGG 6181 GTGATGAATT TTCTATGAGG AGGGTAGAGG TGTCTGGAGT CTCTGTGATG GCTGCCTTTC 6241 GGGCCGCTTG GTCAGCCATC CGGTTGCCTC TAGCCTCGGC GCTGTGTCCC TTTTGATGTC 6301 CTGGACAATG GATTATGCTA AGTCTTTTGG GCAGAAAGAG GGCTTTTAGT AGGGCCAAGA 6361 TCTCGTCTTT ATTTTTGATC TCTTTGCCTT CTGATGTGAG CAACCCACGC CTTCTGTATA 6421 TTTCTCCATG GATATGGGCA GTAGCAAAAG CATAACGGCT ATCAGTATAA ACATTTAGCT 6481 TCTTACCTTC TGCCATCTTT AGGGCCTGGG TGAGTGCTAT CAGTTCAGCC CGCTGAGCGG 6541 ATGTCCCGGC TGGCAGGGCT TTAGCCCAGA TTACCTCGGT CTCGGTGGTC ACCGCAGCTC 6601 CCGCCTTACG CTGTCCCTCT TGTAAGAGAC TGCTTCCATC CGTGTACCAG GTGTGGTCGG 6661 CGTCTGGGAG CGGCTGGTCC GTTAGGTCGG GTCGGGTTCC GTGGGCTTCG GCCAGGATAT 6721 CAAGGCAGTT GTGTTGCAGC CCTTCCTCAG GCAGTGGGAG CAGCGTAGCC GGGTTCAGGG 6781 CTACCACCGG TCCGAACTGG ACCCGGTCCG TGTCCAAAAG CAAGGCCTGA TAGTGAGTCA 6841 TCCGGGCGTT GGAAAGCCAG CGGTCGGGGG GTTGTTTGAC TAGTGCCTCT ACTGCATGGG 6901 GGGCCAGAAT GACTAGTGGC TGTCCCATGG TTAGCTTGCC TGCATCCTTT GTCAGTACGG 6961 CAATGGCTGC TACCATCCGT AGGCAAGGGG GCCACCCAGC TGCTACTGGG TCTAGCTTTT 7021 TGGACAGGTA GGCCACCGGC CGACGCCAAG GTCCCAGTTT TTGCGTTAGG ACACCTTTGG 7081 CGTAGCCCTG CTTCTCGTCG ACAAAGAGTT CAAAGGGCTT AGTCAAATCT GGCAACCCCA 7141 GGGCTGGGGC AGTTAGAAGA GCTTGCTTGA TTTCTTGATA GGCCTTTTGT TGGTCTGGGC 7201 CCCAATTAAA CAGAGTCCCC GTTTTGGTGA GAGGGTACAA GGGGGCTGCC ATTTCTGCAA 7261 ACCCAGGGAT CCAGAGGCGA CAGAAGCCTG CCGTCCCTAG GAACTCCCTT AGTTGTCGAG 7321 GGGTCTTCGG AGTAGGCTGC CCCATCACAG TCTCTTTTCT GGCCTCAGTC AGCCATCTCT 7381 GACCCTCTTT TAGAAGATAC CCCAGATACT TGACCTGTTT CTGGCAAATT TGGGCTTTCT 7441 TGGCCGAGGC CCGATACCCG AGGTTCCCTA GGGTTTGTAA CAGGGCCCGA GTACCTTGTT 7501 GGCAGTCTAG CTCAGAAGTG GCGGCCAGCA GTAAGTCATC CACGTACTGT AGCAGGATCA 7561 AGTCTGGGTG CTGGATCCGG AAGTCTGCTA GGTCTCTGTG CAGTGCCTCA TCAAACAGGG 7621 TGGGACTGTT TTTGAAACCC TGTGGGAGTC TGGTCCAGGT CAATTGTCCT GAGATTCCCA 7681 TCTCTGGATC TCTCCACTCA AAGGCGAAGA GAGGCTGACT GGTGGGGTGG AGTCTCAGGC 7741 AGAAAAAGGC ATCCTTTAAA TCAAGCACAG TGTACCACTG GTGGGACGGT GGGAGCCCGC 7801 TCAAGAGGTT GTAAGGGTTG GGCACGGTGG GGTGGATGTC TTCCACCCGC TTGTTGACTT 7861 CTCTCAGATC CTGGACAGGC CTATAATCAT TAGTCCCTGG TTTCTTAACG GGTAGCAGGG 7921 GCGTGTTCCA GGGGGACTGG CAGGGTACCA GTATTCCCTG GTCCAACAGT CTCTGTATGT 7981 GGGGCTTGAT CCCCAGTCTG GCTTCTTGTG ACATGGGGTA TTGTTTTATG GACACGGGGG 8041 TAGAGGTTGC TTTCAGAGGT ATGATCAGAG GAGCTTGGCG AACTGCCAGT CCCATGCCCC 8101 CGGTTTCCGC CCAGGCCTGA GGAAAATCAG ACAGCCATGT GGACCCTAGA GAAACATCTG 8161 GCTCTTTTGA GGTCTCATGT AGCCGATACT CATCTTCTAT ATTTAGGGTC AACACTTGCA 8221 GGGGCTGCCC CATTGGTCCC ATAACCTGAG CTCCTGATCC CTCAAAGTGG ATTTGGGCTT 8281 TTAGTTTAGT CAGCAAATCT CTTCCTAACA GAGGATAGGG ACAGTCTGGT ACATGGAGGA 8341 AAGAGTGGGT GACCTTACCG GTAGCTAGAT GTACTTTGCG ATCCGTGGTC CAGCGATACC 8401 GCTTTCCTCC AGTAGCCCCT TGGACCCAGG CAGACTTATC ACTTAGGGGT CCAGGATTTT 8461 GGGTCAGCAC GGAGTGTTGG GCCCCAGTAT CTACCAGGAA GGTGACGGGT TGCCCCCCGA 8521 CTTTGAGGGT TATCCTGGGT TCAGGGGGGG GCTCCTGACC CTGACCTCCC TAGTCATCTA 8581 GGGTCAGGAG GGAGGTCTGG GGTCTTGGTC CCCGAGGTCC TCGTGGTTTC TTGGGACAAT 8641 CTTTAGCCCA GTGCCCCTTT TCTTTGCAGT AGGCACACTG GTCGCGATCG AGTTGGGACC 8701 TCCTTCGTTC TCCTCCCTGT CTATCCTGTT TCTGTCCACT AACGACAGTG GCCAATAGCT 8761 TGCTCATCTC TCTATGTCTC CTACGATCTC TTTCTTTCTC TTTCTGCTCA TCCTCTGTCC 8821 TACGGCGTTC TTCTTTTTCC TCTGTTTCTC TCCTGATACG TTCCTCTCTT TCTTCCGGGG 8881 TTTCTCGTTT ATTAAAGATC TTTTCTGCCT CTCTAACCAA ATCTCCAAGC GTCTTGTTTT 8941 TTAAATCTTC TAACCTCTCT AACTTTCTCC CAATGTCTGG GGCAGACTGC CAAATGAAAG 9001 ACATAGACAC ATTAGTTTCT TGCCCTGGGT CCTCAGGGTC ATAAGGAGTG TACCTGCGAT 9061 AGGCTTCCTT AAGTCTCTCT AGGAAGGCCG AGGGAGACTC ATTGGGCCCT TGTGTTATTC 9121 CTTTTACCTT GGCCAAATTG GTGGGGCTTC TGCCCGCGTT TTGGAGACCC GCTAGGAGCA 9181 ACTGGCGATA GTGGACTAGG TGGTTCCTAC CTGCCTGGGT GGTGTAATCC CAGTCTGGGC 9241 GCTCGAGGGG AAAAGCGGCA TCGACTTCAT TGGGCAGTTG AGTGGGGCGC CCATCATCGC 9301 CCCGCACCGC CTTTCTAGCC TCTAAGAGCA CCCGTTGTTT TTCTTCTCCG GTCAGCAGAG 9361 TCCCCAACAG CTGCTGACAG TCGTCCCAGG TGGGCTGATG GGTGATGAGG ACAGACTCGA 9421 TCAGAGCTGT CAGTTTACCT GGATCTTCAG AAAAAGAAGG GTTATTATTT TTCCAGTTGT 9481 AAAGGTCAGA AGAGGAGAAC GGCCAGTATT GAAGCTGTCC GTTTCCTCCT GCGCGGAGGG 9541 GGAATGCCTG CGAGGTAGTG GAGTCGGCCA CAGGGGGCTC CCGTCTCCCA CGTAGGCGAG 9601 ATGCCATTGG GGAGGGGTCC GGTGCCTCTC CCGCAGGGGT CGCTTCTCCA CCATTTCCGT 9661 CCCTGTCGGA AGGGGGTGGT CTTGGGTCCC TATAAGGCGG GGGGTCTTCT GTAAGTAGGT 9721 CGATGAGCGG CCCCCCACTG TCAGAAAGAA CTTGAGGTTT AGGTTTGGCG CCTAGAGAAG 9781 GAGTGAGGGC TGGATAAAGG GAGGATCGAG GCGGGGTCGA ACGAGGAGGT TCAAGGGGGA 9841 GAGACGGGGC GGATGGAGGA AGAGGAGGCG GAGGCTTAGG GTGTACAAAG GGCTTGACCC 9901 AGGGAGGGGG GTCAAAAGCC AAGGCTTCCC AGGTCACGAT GTAGGGGACC TGGTCTGGGT 9961 GTCCATGCGG GCCAGGTGAA AAGACCTTGA TCTTAACCTG GGTGATGAGG TCTCGGTTAA 10021 AGGTGCCGTC TCGCGGCCAT CCGACGTTAA AGGTTGGCCA TTCTGCAGAG CAGAAGGTAA 10081 CCCAACGTCT CTTCTTGACA TCTACCGACT GGTTGTGAGC GATCCGCTCG ACATCTTTCC 10141 AGTGACCTAA GGTCAAACTT AAGGGAGTGG TAACAGTCTG GCCCATATTT TCAGACAAAT 10201 ACAGAAACAC AGTCAGACAG AGACAACACA GAACGATGCC CAAGCTTGGC CAGCTTACCT 10261 CCCGGTGGTG GGTCGGTGGT CCCTGGGCAG GGGTCTCCCG ATCCCGGACG AGCCCCCAAA 10321 TGAAAGACCC CCGCTGACGG GTAGTCAATC ACTCAGAGGA GACCCTCCCA AGGAACAGCG 10381 AGACCACAAG TCGGATGCAA CTGCAAGAGG GTTTATTGGA TACACGGGTA CCCGGGCGAC 10441 TCAGTCAATC GGAGGACTGG CGCGCCGAGT GAGGGGTTGT GGGCTCTTTT ATTGAGCTCT 10501 GCTTATATAG ACCTCCCACC GTACACGCCT ACCGCCCATT TGCGTCAATG GGGCGGAGTT 10561 GTTACGACAT TTTGGAAAGT CCCGTTGATT TTGGTGCCAA AACAAACTCC CATTGACGTC 10621 AATGGGGTGG AGACTTGGAA ATCCCCGTGA GTCAAACCGC TATCCACGCC CATTGATGTA 10681 CTGCCAAAAC CGCATCACCA TGGTAATAGC GATGACTAAT ACGTAGATGT ACTGCCAAGT 10741 AGGAAAGTCC CATAAGGTCA TGTACTGGGC ATAATGCCAG GCGGGCCATT TACCGTCATT 10801 GACGTCAATA GGGGGCGTAC TTGGCATATG ATACACTTGA TGTACTGCCA AGTGGGCAGT 10861 TTACCGTAAA TACTCCACCC ATTGACGTCA ATGGAAAGTC CCTATTGGCG TTACTATGGG 10921 AACATACGTC ATTATTGACG TCAATGGGCG GGGGTCGTTG GGCGGTCAGC CAGGCGGGCC 10981 ATTTACCGTA AGTTATGTAA CGCGGAACTC CATATATGGG CTATGAACTA ATGACCCCGT 11041 AATTGATTAC TATTAATAAC TAGTCAATAA TCAATGTCAA CGCGCTGGCG TAATAGCGAA 11101 GAGGCCCGCA CCGATCGCCC TTCCCAACAG TTGCGCAGCC TGAATGGCGA ATGGCGCCTG 11161 ATGCGGTATT TTCTCCTTAC GCATCTGTGC GGTATTTCAC ACCGCATATG GTGCACTCTC 11221 AGTACAATCT GCTCTGATGC CGCATAGTTA AGCCAGCCCC GACACCCGCC AACACCCGCT 11281 GACGCGCCCT GACGGGCTTG TCTGCTCCCG GCATCCGCTT ACAGACAAGC TGTGACCGTC 11341 TCCGGGAGCT GCATGTGTCA GAGGTTTTCA CCGTCATCAC CGAAACGCGC GAGACGAAAG 11401 GGCCTCGTGA TACGCCTATT TTTATAGGTT AATGTCATGA TAATAATGGT TTCTTAGACG 11461 TCAGGTGGCA CTTTTCGGGG AAATGTGCGC GGAACCCCTA TTTGTTTATT TTTCTAAATA 11521 CATTCAAATA TGTATCCGCT CATGAGACAA TAACCCTGAT AAATGCTTCA ATAATATTGA 11581 AAAAGGAAGA GTATGAGTAT TCAACATTTC CGTGTCGCCC TTATTCCCTT TTTTGCGGCA 11641 TTTTGCCTTC CTGTTTTTGC TCACCCAGAA ACGCTGGTGA AAGTAAAAGA TGCTGAAGAT 11701 CAGTTGGGTG CACGAGTGGG TTACATCGAA CTGGATCTCA ACAGCGGTAA GATCCTTGAG 11761 AGTTTTCGCC CCGAAGAACG TTTTCCAATG ATGAGCACTT TTAAAGTTCT GCTATGTGGC 11821 GCGGTATTAT CCCGTATTGA CGCCGGGCAA GAGCAACTCG GTCGCCGCAT ACACTATTCT 11881 CAGAATGACT TGGTTGAGTA CTCACCAGTC ACAGAAAAGC ATCTTACGGA TGGCATGACA 11941 GTAAGAGAAT TATGCAGTGC TGCCATAACC ATGAGTGATA ACACTGCGGC CAACTTACTT 12001 CTGACAACGA TCGGAGGACC GAAGGAGCTA ACCGCTTTTT TGCACAACAT GGGGGATCAT 12061 GTAACTCGCC TTGATCGTTG GGAACCGGAG CTGAATGAAG CCATACCAAA CGACGAGCGT 12121 GACACCACGA TGCCTGCAGC AATGGCAACA ACGTTGCGCA AACTATTAAC TGGCGAACTA 12181 CTTACTCTAG CTTCCCGGCA ACAATTAATA GACTGGATGG AGGCGGATAA AGTTGCAGGA 12241 CCACTTCTGC GCTCGGCCCT TCCGGCTGGC TGGTTTATTG CTGATAAATC TGGAGCCGGT 12301 GAGCGTGGGT CTCGCGGTAT CATTGCAGCA CTGGGGCCAG A //
pCL-Eco载体图谱质粒图谱pdf版和pCL-Eco载体序列质粒序列等相关资料下载
pCL-Eco质粒载体应用举例
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