LOCUS       Exported                6657 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV production.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-EF1a-CDreV
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6657)
  AUTHORS   Yao S, Yuan P, Ouellette B, Zhou T, Mortrud M, Balaram P, Chatterjee
            S, Wang Y, Daigle TL, Tasic B, Kuang X, Gong H, Luo Q, Zeng S, 
            Curtright A, Dhaka A, Kahan A, Gradinaru V, Chrapkiewicz R, 
            Schnitzer M, Zeng H, Cetin A
  TITLE     RecV recombinase system for in vivo targeted optogenomic 
            modifications of single cells or cell populations.
  JOURNAL   Nat Methods. 2020 Mar 23. pii: 10.1038/s41592-020-0774-3. doi: 
            10.1038/s41592-020-0774-3.
  PUBMED    32203389
REFERENCE   2  (bases 1 to 6657)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6657
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   27..156
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        257..1435
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          488..1426
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1383..1403
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    2810..3398
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2863..2883)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3281..3292)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3430..3906
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3629..3648)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3946..4086
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3946..4075
                     /label=AAV2 ITR
     rep_origin      4161..4616
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4248..4267)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4458..4479
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4633..4652)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4752..4774
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4812..4830)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4898..5002
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5003..5863
                     /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(5221..5240)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6034..6622
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6523..6542
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cttttgctgg ccttttgctc acatgtcctg caggcagctg cgcgctcgct cgctcactga
       61 ggccgcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca
      121 gagagggagt ggccaactcc atcactaggg gttcctgcgg ccgcacgcgt aagctttgca
      181 aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct tctaggtctt
      241 gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
      301 ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg
      361 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
      421 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
      481 aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg gttatggccc
      541 ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc ccgagcttcg
      601 ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc
      661 ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg
      721 cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc
      781 gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc acactggtat
      841 ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc
      901 gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg
      961 gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct
     1021 ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctgcagg
     1081 gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag
     1141 gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc
     1201 gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag gttgggggga
     1261 ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc
     1321 ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat
     1381 tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgaggtac
     1441 cggatcctct agagtcgacg ccgccaccat gcatacactg tatgcccctg gaggatatga
     1501 tattatggga tatctgattc agattatgaa aagacctaat cctcaggtgg aactgggacc
     1561 tgtggataca agcgtggccc tgattctgtg tgatctgaaa cagaaagata cacctattgt
     1621 gtatgccagc gaagcctttc tgtatatgac aggatatagc aatgccgaag tgctgggaag
     1681 aaattgtaga tttctgcaga gccctgatgg aatggtgaaa cctaaaagca caagaaaata
     1741 tgtggatagc aatacaatta atacaatgag aaaagccatt gatagaaatg ccgaagtgca
     1801 ggtggaagtg gtgaatttta aaaaaaatgg acagagattt gtgaattttc tgacaatgat
     1861 tcctgtgaga gatgaaacag gagaatatag atatagcatg ggatttcagt gtgaaacaga
     1921 aggaggcggt ggaggtcagc ttgcatggtt tccaattagt cctgaaatgg cacgcgaata
     1981 ctttcttcag ttgcacgatg cagacctggc ctccactacc atcgacaagc actatgctat
     2041 gcttaatatg cttctgtccc actgcggact gccacccttg tccgacgaca agtcagtgag
     2101 tcttgccatg agaagaatta gaagagaagc cgcaaccgaa aagggtgaga ggacaggaca
     2161 ggcaatcccc ctgcgctggg acgacctgaa gctgctggat gtgctgctca gcaggagcga
     2221 gcggctggtc gacctgcgca acagggcttt cctgttcgta gcctataaca ccctcatgag
     2281 aatgtctgaa atatcacgca tcagggttgg ggacttggat cagacaggag acacagtgac
     2341 cctgcacatc agtcacacta agacaatcac cacagctgcg ggccttgaca aagtgctctc
     2401 ccggcgaacc acagcagtgc tcaatgactg gctggacgtc agtgggctta gagaacatcc
     2461 agacgctgtg ctcttcccac ctatacaccg gtcaaacaaa gcccgcatta ctaccacgcc
     2521 cctgaccgcc cctgccatgg agaagatttt cagtgatgcc tgggtgctgc tgaacaaacg
     2581 ggacgccacc cccaataaag ggaggtatag gacctggacc ggccattccg ccagggtggg
     2641 tgccgcaata gacatggccg agaaacaggt gtctatggtc gagattatgc aggaagggac
     2701 atggaagaag cctgaaacac tgatgcggta tctcagaagg ggcggagtgt ccgtgggagc
     2761 caattctcga ctgatggata gctaagaatt cgatatcaag cttatcgata atcaacctct
     2821 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     2881 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2941 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     3001 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     3061 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3121 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3181 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctatgttgc
     3241 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3301 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3361 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag cgctgctcga
     3421 gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt
     3481 gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac
     3541 taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt
     3601 gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca
     3661 caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct
     3721 cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg
     3781 tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc
     3841 tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct
     3901 gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga
     3961 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     4021 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     4081 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     4141 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4201 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     4261 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4321 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     4381 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4441 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     4501 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     4561 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     4621 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     4681 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     4741 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     4801 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     4861 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4921 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4981 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5041 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5101 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     5161 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     5221 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     5281 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5341 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5401 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5461 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5521 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     5581 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5641 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5701 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5761 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5821 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     5881 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5941 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6001 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6061 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6121 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6181 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6241 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6301 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6361 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6421 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6481 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6541 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6601 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggc
//
