| 质粒名称: | SFG.CNb30_opt.IRES.eGFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 22493 |
| 存储实验室: | Martin Pule |
|
载体骨架基本信息 |
|
| 载体名称: | SFG |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 7678 bp |
| 载体类型: | 逆转录病毒 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Codon optimised Calcineurin B mutant 30,CNb30,CNb |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 519 bp |
| 基因突变: | L124T; K125-LA-Ins |
| GenBank编号 | GQ463597 |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | NcoI (未破坏) |
| 3' 克隆位点: | MluI (未破坏) |
| 5' 测序引物 | ttacacagtcctgctgaccacc |
| 3' 测序引物 | caagcggcttcggccagtaac |
|
文献引用方法 |
|
| 材料和方法部分: | SFG.CNb30_opt.IRES.eGFP was a gift from Martin Pule (Addgene plasmid # 22493 ; http://n2t.net/addgene:22493 ; RRID:Addgene_22493) |
| 参考文献部分: | Generation of EBV-specific cytotoxic T-cells that are resistant to calcineurin inhibitors for the treatment of post-transplant lymphoproliferative disease. Brewin J, Mancao C, Straathof K, Karlsson H, Samarasinghe S, Amrolia PJ, Pule M. Blood. 2009 Sep 21. ():. 10.1182/blood-2009-07-228387 PubMed 19770360 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS SFG.CNb30_opt.IRES.eGFP 8197 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS SFG.CNb30_opt.IRES.eGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8197)
AUTHORS Brewin J, Mancao C, Straathof K, Karlsson H, Samarasinghe S, Amrolia
PJ, Pule M
TITLE Generation of EBV-specific cytotoxic T-cells that are resistant to
calcineurin inhibitors for the treatment of post-transplant
lymphoproliferative disease.
JOURNAL Blood. 2009 Sep 21. ():.
PUBMED 19770360
REFERENCE 2 (bases 1 to 8197)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8197
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 655..1012
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
primer_bind 1423..1445
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 1461..1477
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
misc_feature 1503..1877
/label=pol region
/note="Moloney murine leukemia virus (MMLV) pol region
containing the splice acceptor site"
primer_bind 1815..1831
/label=pBMN 5'
/note="MMLV sequence, for inserts in pBMN retroviral
vector"
regulatory 1877..1886
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
misc_feature 2466..3017
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(2610..2627)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 2837..2856
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 3017..3736
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 3017..3733
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(3062..3083)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(3323..3342)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 3670..3691
/label=EGFP-C
/note="EGFP, forward primer"
LTR 4002..4471
/note="long terminal repeat from Moloney murine leukemia
virus"
primer_bind complement(5170..5187)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(5170..5186)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5179..5201)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(5395..5414)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5514..5536
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(5575..5593)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5661..5765
/gene="bla"
/label=AmpR promoter
CDS 5766..6626
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(5984..6003)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6797..7385
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7286..7305
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7539..7556
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 7673..7694
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 7709..7739
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7747..7763
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 7752..7774
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 7771..7787
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 7771..7787
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
LTR 8196..592
/note="long terminal repeat from Moloney murine leukemia
virus"
ORIGIN
1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
61 ggaaaaatac ataactgaga atagaaaagt tcagatcaag gtcaggaaca gatggaacag
121 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
181 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
241 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
301 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
361 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttatgctccc cgagctcaat
421 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
481 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
541 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct
601 cgtccgggat cgggagaccc ctgcccaggg accaccgacc caccaccggg aggtaagctg
661 gccagcaact tatctgtgtc tgtccgattg tctagtgtct atgactgatt ttatgcgcct
721 gcgtcggtac tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga
781 gttcggaaca cccggccgca accctgggag acgtcccagg gacttcgggg gccgtttttg
841 tggcccgacc tgagtcctaa aatcccgatc gtttaggact ctttggtgca ccccccttag
901 aggagggata tgtggttctg gtaggagacg agaacctaaa acagttcccg cctccgtctg
961 aatttttgct ttcggtttgg gaccgaagcc gcgccgcgcg tcttgtctgc tgcagcatcg
1021 ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc tgaaaatatg ggcccgggct
1081 agcctgttac cactccctta agtttgacct taggtcactg gaaagatgtc gagcggatcg
1141 ctcacaacca gtcggtagat gtcaagaaga gacgttgggt taccttctgc tctgcagaat
1201 ggccaacctt taacgtcgga tggccgcgag acggcacctt taaccgagac ctcatcaccc
1261 aggttaagat caaggtcttt tcacctggcc cgcatggaca cccagaccag gtggggtaca
1321 tcgtgacctg ggaagccttg gcttttgacc cccctccctg ggtcaagccc tttgtacacc
1381 ctaagcctcc gcctcctctt cctccatccg ccccgtctct cccccttgaa cctcctcgtt
1441 cgaccccgcc tcgatcctcc ctttatccag ccctcactcc ttctctaggc gcccccatat
1501 ggccatatga gatcttatat ggggcacccc cgccccttgt aaacttccct gaccctgaca
1561 tgacaagagt tactaacagc ccctctctcc aagctcactt acaggctctc tacttagtcc
1621 agcacgaagt ctggagacct ctggcggcag cctaccaaga acaactggac cgaccggtgg
1681 tacctcaccc ttaccgagtc ggcgacacag tgtgggtccg ccgacaccag actaagaacc
1741 tagaacctcg ctggaaagga ccttacacag tcctgctgac cacccccacc gccctcaaag
1801 tagacggcat cgcagcttgg atacacgccg cccacgtgaa ggctgccgac cccgggggtg
1861 gaccatcctc taggctgcca ccatgggcaa cgaggccagc taccctctgg agatgtgctc
1921 ccacttcgac gccgacgaga tcaagcggct gggcaagcgc ttcaagaagc tggacctgga
1981 caacagcggc agcctgagcg tggaggagtt tatgtctctg cccgagctgc agcagaaccc
2041 cctggtgcag cgcgtgatcg acatcttcga caccgacggc aacggcgagg tggacttcaa
2101 ggagttcatc gagggcgtga gccagttcag cgtgaagggc gacaaggagc agaagctgcg
2161 gttcgccttc cggatctacg atatggataa agatggctat atttctaatg gcgagctgtt
2221 ccaggtgctg aagatgatgg tgggcaacaa taccaagctg gccgataccc agctgcagca
2281 gatcgtggac aagaccatca tcaacgccga caaggacggc gacggcagaa tcagcttcga
2341 ggagttctgt gccgtggtgg gaggcctgga tattcacaaa aaaatggtgg tggacgtgtg
2401 aacgcgtacg tcgcgaccgc ggacatgtac agagctcgag cgggatcaat tccgcccccc
2461 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
2521 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
2581 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
2641 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac
2701 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
2761 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
2821 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
2881 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt
2941 ttagtcgagg ttaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga
3001 aaaacacgat aataatatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
3061 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
3121 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
3181 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
3241 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
3301 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
3361 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
3421 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
3481 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
3541 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
3601 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
3661 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
3721 cgagctgtac aagtaatccg gattagtcca atttgttaaa gacaggatat cagtggtcca
3781 ggctctagtt ttgactcaac aatatcacca gctgaagcct atagagtacg agccatagat
3841 aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag
3901 gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata cataactgag
3961 aatagagaag ttcagatcaa ggtcaggaac agatggaaca gctgaatatg ggccaaacag
4021 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gaacagctga
4081 atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac
4141 agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc agatgtttcc
4201 agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc
4261 ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct
4321 cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa
4381 ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag
4441 tgattgacta cccgtcagcg ggggtctttc acacatgcag catgtatcaa aattaatttg
4501 gttttttttc ttaagtattt acattaaatg gccatagtac ttaaagttac attggcttcc
4561 ttgaaataaa catggagtat tcagaatgtg tcataaatat ttctaatttt aagatagtat
4621 ctccattggc tttctacttt ttcttttatt tttttttgtc ctctgtcttc catttgttgt
4681 tgttgttgtt tgtttgtttg tttgttggtt ggttggttaa ttttttttta aagatcctac
4741 actatagttc aagctagact attagctact ctgtaaccca gggtgacctt gaagtcatgg
4801 gtagcctgct gttttagcct tcccacatct aagattacag gtatgagcta tcatttttgg
4861 tatattgatt gattgattga ttgatgtgtg tgtgtgtgat tgtgtttgtg tgtgtgactg
4921 tgaaaatgtg tgtatgggtg tgtgtgaatg tgtgtatgta tgtgtgtgtg tgagtgtgtg
4981 tgtgtgtgtg tgcatgtgtg tgtgtgtgac tgtgtctatg tgtatgactg tgtgtgtgtg
5041 tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg tgtgttgtga aaaaatattc tatggtagtg
5101 agagccaacg ctccggctca ggtgtcaggt tggtttttga gacagagtct ttcacttagc
5161 ttggaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
5221 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
5281 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta
5341 ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat
5401 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
5461 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
5521 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgatgacga aagggcctcg
5581 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
5641 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
5701 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
5761 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
5821 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
5881 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
5941 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
6001 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
6061 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
6121 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
6181 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
6241 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
6301 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
6361 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
6421 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
6481 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
6541 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
6601 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
6661 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
6721 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
6781 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
6841 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
6901 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
6961 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
7021 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
7081 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
7141 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcat tgagaaagcg
7201 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
7261 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
7321 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
7381 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
7441 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
7501 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
7561 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
7621 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
7681 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
7741 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
7801 agctttgctc ttaggagttt cctaatacat cccaaactca aatatataaa gcatttgact
7861 tgttctatgc cctagggggc ggggggaagc taagccagct ttttttaaca tttaaaatgt
7921 taattccatt ttaaatgcac agatgttttt atttcataag ggtttcaatg tgcatgaatg
7981 ctgcaatatt cctgttacca aagctagtat aaataaaaat agataaacgt ggaaattact
8041 tagagtttct gtcattaacg tttccttcct cagttgacaa cataaatgcg ctgctgagca
8101 agccagtttg catctgtcag gatcaatttc ccattatgcc agtcatatta attactagtc
8161 aattagttga tttttatttt tgacatatac atgtgaa
//
