pCAG-T7pol

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pCAGT7pol载体基本信息

质粒名称: pCAG-T7pol 
出品公司: 爱迪基因
目录编号: 59926
存储实验室: Ian Wickersham

载体骨架基本信息
载体名称: pCAGEN
载体抗性: 氨苄青霉素
空载体大小: 4795 bp
完整质粒大小: 7447 bp
载体类型: 哺乳细胞表达
高拷贝/低拷贝: 高拷贝
生长条件: 37 ℃

插入基因信息
插入基因名称: T7 RNA polymerase
物种来源: Bacteriophage T7
插入大小: 2652 bp
启动子 CAG
5' 测序引物 CTCTAGAGCCTCTGCTAACc
3' 测序引物 CAGAAGTCAGATGCTCAAG

文献引用方法
材料和方法部分: pCAG-T7pol was a gift from Ian Wickersham (Addgene plasmid # 59926 ; http://n2t.net/addgene:59926 ; RRID:Addgene_59926)

pCAGT7pol载体图谱质粒图谱和多克隆位点信息

pCAG-T7pol质粒图谱

pCAGT7pol载体简介

pCAGT7pol载体序列

LOCUS       pCAG-T7pol               7452 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  Expression vector for T7 RNA polymerase gene.
ACCESSION   .
VERSION     .
KEYWORDS    pCAG-T7pol
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7452)
  TITLE     Genetic Neuroengineering Group Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7452)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7452
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1673
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1597..1619
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1681..1700
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(4474..4493)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_site      4539..4594
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     primer_bind     complement(4593..4612)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4955..4971)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4955..4971)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4968..4990)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4979..4995
                     /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)."
     promoter        complement(5003..5033)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5048..5069
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5128..5323
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5174..5309
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5236..5255
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    5329..5463
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5366..5385)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5420..5439
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5531..5548)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5702..6290)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5782..5801)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6461..7321)
                     /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     7084..7103
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7322..7426)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct
     1441 ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc
     1501 ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg
     1561 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg
     1621 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg
     1681 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattcgcc gccaccatga
     1741 acacgattaa catcgctaag aacgacttct ctgacatcga actggctgct atcccgttca
     1801 acactctggc tgaccattac ggtgagcgtt tagctcgcga acagttggcc cttgagcatg
     1861 agtcttacga gatgggtgaa gcacgcttcc gcaagatgtt tgagcgtcaa cttaaagctg
     1921 gtgaggttgc ggataacgct gccgccaagc ctctcatcac taccctactc cctaagatga
     1981 ttgcacgcat caacgactgg tttgaggaag tgaaagctaa gcgcggcaag cgcccgacag
     2041 ccttccagtt cctgcaagaa atcaagccgg aagccgtagc gtacatcacc attaagacca
     2101 ctctggcttg cctaaccagt gctgacaata caaccgttca ggctgtagca agcgcaatcg
     2161 gtcgggccat tgaggacgag gctcgcttcg gtcgtatccg tgaccttgaa gctaagcact
     2221 tcaagaaaaa cgttgaggaa caactcaaca agcgcgtagg gcacgtctac aagaaagcat
     2281 ttatgcaagt tgtcgaggct gacatgctct ctaagggtct actcggtggc gaggcgtggt
     2341 cttcgtggca taaggaagac tctattcatg taggagtacg ctgcatcgag atgctcattg
     2401 agtcaaccgg aatggttagc ttacaccgcc aaaatgctgg cgtagtaggt caagactctg
     2461 agactatcga actcgcacct gaatacgctg aggctatcgc aacccgtgca ggtgcgctgg
     2521 ctggcatctc tccgatgttc caaccttgcg tagttcctcc taagccgtgg actggcatta
     2581 ctggtggtgg ctattgggct aacggtcgtc gtcctctggc gctggtgcgt actcacagta
     2641 agaaagcact gatgcgctac gaagacgttt acatgcctga ggtgtacaaa gcgattaaca
     2701 ttgcgcaaaa caccgcatgg aaaatcaaca agaaagtcct agcggtcgcc aacgtaatca
     2761 ccaagtggaa gcattgtccg gtcgaggaca tccctgcgat tgagcgtgaa gaactcccga
     2821 tgaaaccgga agacatcgac atgaatcctg aggctctcac cgcgtggaaa cgtgctgccg
     2881 ctgctgtgta ccgcaaggac aaggctcgca agtctcgccg tatcagcctt gagttcatgc
     2941 ttgagcaagc caataagttt gctaaccata aggccatctg gttcccttac aacatggact
     3001 ggcgcggtcg tgtttacgct gtgtcaatgt tcaacccgca aggtaacgat atgaccaaag
     3061 gactgcttac gctggcgaaa ggtaaaccaa tcggtaagga aggttactac tggctgaaaa
     3121 tccacggtgc aaactgtgcg ggtgtcgata aggttccgtt ccctgagcgc atcaagttca
     3181 ttgaggaaaa ccacgagaac atcatggctt gcgctaagtc tccactggag aacacttggt
     3241 gggctgagca agattctccg ttctgcttcc ttgcgttctg ctttgagtac gctggggtac
     3301 agcaccacgg cctgagctat aactgctccc ttccgctggc gtttgacggg tcttgctctg
     3361 gcatccagca cttctccgcg atgctccgag atgaggtagg tggtcgcgcg gttaacttgc
     3421 ttcctagtga aaccgttcag gacatctacg ggattgttgc taagaaagtc aacgagattc
     3481 tacaagcaga cgcaatcaat gggaccgata acgaagtagt taccgtgacc gatgagaaca
     3541 ctggtgaaat ctctgagaaa gtcaagctgg gcactaaggc actggctggt caatggctgg
     3601 cttacggtgt tactcgcagt gtgactaagc gttcagtcat gacgctggct tacgggtcca
     3661 aagagttcgg cttccgtcaa caagtgctgg aagataccat tcagccagct attgattccg
     3721 gcaagggtct gatgttcact cagccgaatc aggctgctgg atacatggct aagctgattt
     3781 gggaatctgt gagcgtgacg gtggtagctg cggttgaagc aatgaactgg cttaagtctg
     3841 ctgctaagct gctggctgct gaggtcaaag ataagaagac tggagagatt cttcgcaagc
     3901 gttgcgctgt gcattgggta actcctgatg gtttccctgt gtggcaggaa tacaagaagc
     3961 ctattcagac gcgcttgaac ctgatgttcc tcggtcagtt ccgcttacag cctaccatta
     4021 acaccaacaa agatagcgag attgatgcac acaaacagga gtctggtatc gctcctaact
     4081 ttgtacacag ccaagacggt agccaccttc gtaagactgt agtgtgggca cacgagaagt
     4141 acggaatcga atcttttgca ctgattcacg actccttcgg taccattccg gctgacgctg
     4201 cgaacctgtt caaagcagtg cgcgaaacta tggttgacac atatgagtct tgtgatgtac
     4261 tggctgattt ctacgaccag ttcgctgacc agttgcacga gtctcaattg gacaaaatgc
     4321 cagcacttcc ggctaaaggt aacttgaacc tccgtgacat cttagagtcg gacttcgcgt
     4381 tcgcgtaagc ggccgcactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt
     4441 ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg
     4501 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
     4561 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     4621 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct
     4681 ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc
     4741 tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg
     4801 ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag
     4861 atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc
     4921 cctcgacctg cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     4981 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5041 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5101 cgggaaacct gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc
     5161 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     5221 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     5281 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt
     5341 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     5401 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     5461 atccgctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     5521 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5581 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5641 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5701 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5761 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5821 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5881 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5941 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6001 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6061 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6121 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     6181 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6241 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6301 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6361 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6421 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6481 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6541 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6601 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6661 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6721 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6781 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6841 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6901 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6961 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7021 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7081 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7141 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7201 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7261 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7321 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7381 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7441 aagtgccacc tg
//

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