pAAV-FLEX-GFP

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质粒基本信息

质粒名称: pAAV-FLEX-GFP 
出品公司: 爱迪基因
目录编号: 28304
存储实验室: Edward Boyden

载体骨架基本信息
载体名称: AAV-FLEX
载体出品公司: Scott Sternson
载体抗性: 氨苄霉素
空载体大小: 5047 bp
完整质粒大小: 5767 bp
载体类型: 哺乳细胞表达,AAV, Cre/Lox ; Adeno-associated virus
高拷贝/低拷贝: 低拷贝
生长条件: 30 ℃

插入基因信息
插入基因名称: GFP
物种来源: A. victoria
插入大小: 720 bp
基因又名: Green fluorescent protein
基因突变: N/A
启动子 CAG
克隆方法: 限制性内切酶
5' 克隆位点: KpnI(未破坏)
3' 克隆位点: BsrGI(未破坏)
5' 测序引物 cttctggcgtgtgaccgg

文献引用方法
材料和方法部分: pAAV-FLEX-GFP was a gift from Edward Boyden (爱迪基因 plasmid # 28304)

质粒图谱信息

pAAV-FLEX-GFP质粒图谱

质粒简介

质粒序列

LOCUS       pAAV-FLEX-GFP                5764 bp ds-DNA     circular SYN 10-JAN-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-FLEX-GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5764)
  TITLE     Accessory plasmids for optogenetics
  JOURNAL   Unpublished
COMMENT     This file is created by Vector NTI
            http://www.biofeng.com/
COMMENT     ORIGDB|GenBank
COMMENT     VNTAUTHORNAME|biofeng.com|
COMMENT     VNTNAME|pAAV-FLEX-GFP|
FEATURES             Location/Qualifiers
     source          1..5764
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   56..185
                     /label=AAV2 ITR
     polyA_signal    347..468
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(384..403)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     438..457
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    511..1099
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             617..628
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1026..1046
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    complement(1141..1174)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    complement(1245..1278)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     regulatory      1295..1304
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1301..2020
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1346..1367)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1607..1626)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1954..1975
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    2043..2076
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    2139..2172
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     primer_bind     complement(2207..2226)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(2288..2310)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(2589..2866)
                     /label=chicken beta-actin promoter
     repeat_region   3177..3306
                     /label=AAV2 ITR
     rep_origin      3378..3833
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3465..3484)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3675..3696
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3859..3963
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3964..4824
                     /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(4182..4201)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4995..5583
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5484..5503
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5737..5754
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg
       61 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
      121 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
      181 ttccttgtag ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa
      241 gatccttatc gattttacca catttgtaga ggttttactt gctttaaaaa acctcccaca
      301 tctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
      361 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
      421 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgctc
      481 gaagcggccg cccgggtcga cggtatcgat gcggggaggc ggcccaaagg gagatccgac
      541 tcgtctgagg gcgaaggcga agacgcggaa gaggccgcag agccggcagc aggccgcggg
      601 aaggaaggtc cgctggattg agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa
      661 tccaggtggc aacacaggcg agcagccaag gaaaggacga tgatttcccc gacaacacca
      721 cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga
      781 tgagttccgc cgtggcaata gggaggggga aagcgaaagt cccggaaagg agctgacagg
      841 tggtggcaat gccccaacca gtgggggttg cgtcagcaaa cacagtgcac accacgccac
      901 gttgcctgac aacgggccac aactcctcat aaagagacag caaccaggat ttatacaagg
      961 aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt
     1021 atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt
     1081 tgtaatccag aggttgatta tcgataagct tgatatcgaa ttcgaacctc ttcgagggac
     1141 ataacttcgt ataatgtatg ctatacgaag ttatgggtcg atggtgagat ctggactaga
     1201 gggtcgatgg tgatgcttgg atcaaaatag gaagaatccc aatgataact tcgtataaag
     1261 tatcctatac gaagttatac tagaattcgg taccgccacc atggtgagca agggcgagga
     1321 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     1381 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     1441 catttgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     1501 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     1561 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     1621 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     1681 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     1741 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     1801 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     1861 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     1921 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     1981 cgccgggatc actctcggca tggacgagct gtacaagtaa aactcgaggg gggatcccta
     2041 gtataacttc gtatagcata cattatacga agttatgggt cgatggtgat gcttggcaat
     2101 tcgggtcgat ggtgaagcat tggtcttcct attttgatat aacttcgtat aggatacttt
     2161 atacgaagtt atggggatcc aattctttgc caaaatgatg agacagcaca ataaccagca
     2221 cgttgcccag gagctgtagg aaaaagaaga aggcatgaac atggttagca gaggctctag
     2281 agccgccggt cacacgccag aagccgaacc ccgccctgcc ccgtcccccc cgaaggcagc
     2341 cgtccccccg cggacagccc cgagccgctc cccccgcaca aagggccctc ccggagcccc
     2401 tcaaggcttt cacgcagcca cagaaaagaa acaagccgtc attaaaccaa gcgctaatta
     2461 cagcccggag gagaagggcc gtcccgcccg ctcacctgtg ggagtaacgc ggtcagtcag
     2521 agccggggcg ggcggcgcga ggcggcggcg gagcggggca cggggcgaag gcagcgcgca
     2581 gcgactcccg cccgccgcgc gcttcgcttt ttatagggcc gccgccgccg ccgcctcgcc
     2641 ataaaaggaa actttcggag cgcgccgctc tgattggctg ccgccgcacc tctccgcctc
     2701 gccccgcccc gcccctcgcc ccgccccgcc ccgcctggcg cgcgcccccc cccccccccc
     2761 gcccccatcg ctgcacaaaa taattaaaaa ataaataaat acaaaattgg gggtggggag
     2821 gggggggaga tggggagagt gaagcagaac gtggggctca cctcgaccat ggtaatagcg
     2881 atgactaata cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca
     2941 taatgccagg cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga
     3001 tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa
     3061 tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt caatggtact
     3121 cttcctagag catggctacg tagataagta gcatggcggg ttaatcatta actacaagga
     3181 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     3241 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     3301 gcgcagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     3361 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3421 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3481 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3541 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3601 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3661 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3721 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     3781 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     3841 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3901 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3961 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4021 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4081 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4141 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4201 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4261 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4321 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4381 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     4441 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4501 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4561 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     4621 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4681 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4741 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4801 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     4861 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     4921 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4981 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5041 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5101 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5161 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5221 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5281 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5341 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5401 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5461 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5521 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5581 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5641 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     5701 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     5761 gaag
//

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