LOCUS       Exported                6663 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV production.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-EF1a-CCreV
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6663)
  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 6663)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6663
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1400
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1357..1377
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    2790..3378
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2843..2863)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3261..3272)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3410..3886
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3609..3628)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3926..4066
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3926..4055
                     /label=AAV2 ITR
     rep_origin      4141..4596
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4228..4247)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4438..4459
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4613..4632)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4732..4754
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4792..4810)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4878..4982
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4983..5843
                     /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(5201..5220)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6014..6602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6503..6522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc gacgccgcca
     1441 ccatgcatac actgtatgcc cctggaggat atgatattat gggatatctg attcagatta
     1501 tgaaaagacc taatcctcag gtggaactgg gacctgtgga tacaagcgtg gccctgattc
     1561 tgtgtgatct gaaacagaaa gatacaccta ttgtgtatgc cagcgaagcc tttctgtata
     1621 tgacaggata tagcaatgcc gaagtgctgg gaagaaattg tagatttctg cagagccctg
     1681 atggaatggt gaaacctaaa agcacaagaa aatatgtgga tagcaataca attaatacaa
     1741 tgagaaaagc cattgataga aatgccgaag tgcaggtgga agtggtgaat tttaaaaaaa
     1801 atggacagag atttgtgaat tttctgacaa tgattcctgt gagagatgaa acaggagaat
     1861 atagatatag catgggattt cagtgtgaaa cagaaggagg cggtggaggt aaccggaaat
     1921 ggtttcccgc agaacctgaa gatgttcgcg attatcttct atatcttcag gcgcgcggtc
     1981 tggcagtaaa aactatccag caacatttgg gccagctaaa catgcttcat cgtcggtccg
     2041 ggctgccacg accaagtgac agcaatgctg tttcactggt tatgcggcgg atccgaaaag
     2101 aaaacgttga tgccggtgaa cgtgcaaaac aggctctagc gttcgaacgc actgatttcg
     2161 accaggttcg ttcactcatg gaaaatagcg atcgctgcca ggatatacgt aatctggcat
     2221 ttctggggat tgcttataac accctgttac gtatagccga aattgccagg atcagggtta
     2281 aagatatctc acgtactgac ggtgggagaa tgttaatcca tattggcaga acgaaaacgc
     2341 tggttagcac cgcaggtgta gagaaggcac ttagcctggg ggtaactaaa ctggtcgagc
     2401 gatggatttc cgtctctggt gtagctgatg atccgaataa ctacctgttt tgccgggtca
     2461 gaaaaaatgg tgttgccgcg ccatctgcca ccagccagct atcaactcgc gccctggaag
     2521 ggatttttga agcaactcat cgattgattt acggcgctaa ggatgactct ggtcagagat
     2581 acctggcctg gtctggacac agtgcccgtg tcggagccgc gcgagatatg gcccgcgctg
     2641 gagtttcaat accggagatc atgcaagctg gtggctggac caatgtaaat attgtcatga
     2701 actatatccg taacctggat agtgaaacag gggcaatggt gcgcctgctg gaagatggcg
     2761 attaagaatt cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     2821 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     2881 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     2941 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     3001 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     3061 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     3121 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     3181 ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt ctgcgcggga
     3241 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     3301 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     3361 tttgggccgc ctccccgcat cgataccgag cgctgctcga gagatctacg ggtggcatcc
     3421 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag
     3481 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat
     3541 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc
     3601 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat
     3661 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca
     3721 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat
     3781 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa
     3841 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt
     3901 aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     3961 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     4021 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     4081 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     4141 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4201 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4261 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4321 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     4381 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4441 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     4501 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     4561 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     4621 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     4681 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     4741 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     4801 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     4861 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4921 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4981 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     5041 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     5101 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5161 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5221 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5281 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5341 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5401 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5461 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5521 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5581 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5641 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5701 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5761 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5821 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5881 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5941 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6001 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6061 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6121 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6181 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6241 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6301 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6361 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6421 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6481 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6541 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6601 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6661 tgt
//
