LOCUS       Exported                4595 bp ds-DNA     circular SYN 12-MAY-2023
DEFINITION  AAV vector for cardiomyocyte specific expression of Emerald Green 
            Fluorescent Protein using cardiac troponin T promoter.
ACCESSION   .
VERSION     .
KEYWORDS    011LP-pAAV-TNT4-EmGFP-pA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4595)
  AUTHORS   Ni L, Scott L Jr, Campbell HM, Pan X, Alsina KM, Reynolds J, 
            Philippen LE, Hulsurkar M, Lagor WR, Li N, Wehrens XHT
  TITLE     Atrial-Specific Gene Delivery Using an Adeno-Associated Viral 
            Vector.
  JOURNAL   Circ Res. 2019 Jan 18;124(2):256-262. doi: 
            10.1161/CIRCRESAHA.118.313811.
  PUBMED    30582449
REFERENCE   2  (bases 1 to 4595)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2023 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4595
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   4..133
                     /label=AAV2 ITR
     protein_bind    791..815
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             827..892
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             896..922
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             944..1663
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(989..1010)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1250..1269)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1597..1618
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1687..1808
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1698..1717)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1752..1771
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   1956..2085
                     /label=AAV2 ITR
     primer_bind     complement(2151..2168)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2322..2910)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2402..2421)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3081..3941)
                     /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     3704..3723
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3942..4046)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4072..4527)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4209..4230)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4421..4440
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
       61 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
      121 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac atccttaagc
      181 tcagtccatt aggagccagt agcctggaag atgtctttac ccccagcatc agttcaagtg
      241 gagcagcaca taactcttgc cctctgcctt ccaagattct ggtgctgaga cttatggagt
      301 gtcttggagg ttgccttctg ccccccaacc ctgctcccag ctggccctcc caggcctggg
      361 ttgctggcct ctgctttatc aggattctca agagggacag ctggtttatg ttgcatgact
      421 gttccctgca tatctgctct ggttttaaat agcttatctg agcagctgga ggaccacatg
      481 ggcttatatg gcgtggggta catgttcctg tagccttgtc cctggcacct gccaaaatag
      541 cagccaacac cccccacccc caccgccatc cccctgcccc acccgtcccc tgtcgcacat
      601 tcctccctcc gcagggctgg ctcaccaggc cccagcccac atgcctgctt aaagccctct
      661 ccatcctctg cctcacccag tccccgctga gactgagcag acgcctccag gatctgtcgg
      721 caggaattcc agaccctcta gaaaatatcc aaggatctgg atcctcccaa gctggctagt
      781 taagctatca acaagtttgt acaaaaaagc aggctttaaa accatggact acaaagacca
      841 tgacggtgat tataaagatc atgacatcga ttacaaggat gacgatgaca agctttaccc
      901 atacgatgtt ccagattacg ctacgcgtgg aggcgctggc ggaatggtga gcaagggcga
      961 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     1021 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     1081 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcac
     1141 ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
     1201 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     1261 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     1321 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     1381 caacagccac aaggtctata tcaccgccga caagcagaag aacggcatca aggtgaactt
     1441 caagacccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     1501 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     1561 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     1621 cgccgccggg atcactctcg gcatggacga gctgtacaag taagcggccg cttcgagcag
     1681 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     1741 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     1801 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg
     1861 aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgagagc atggctacgt
     1921 agataagtag catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc
     1981 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     2041 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgca ttaatgaatc
     2101 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     2161 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2221 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2281 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2341 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2401 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2461 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2521 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2581 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2641 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2701 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2761 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2821 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2881 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2941 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3001 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3061 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3121 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3181 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3241 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3301 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3361 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3421 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3481 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3541 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3601 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3661 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3721 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3781 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3841 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3901 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3961 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4021 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
     4081 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     4141 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     4201 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     4261 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     4321 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     4381 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     4441 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     4501 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg
     4561 aagggcgatc ggtgcgggcc tcttcgctat tacgc
//
