LOCUS       sequence-393067-        4595 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for cardiomyocyte specific expression of Emerald Green 
            Fluorescent Protein using cardiac troponin T promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1161/CIRCRESAHA.118"; journalName: "Circ Res"; date: 
            "2019-01-18- 18"; volume: "124"; issue: "2"; pages: "256-262"
COMMENT     Sequence Label: 011LP-pAAV-TNT4-EmGFP-pA
FEATURES             Location/Qualifiers
     source          1..4595
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   248..377
                     /label=AAV2 ITR
     polyA_signal    525..646
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(562..581)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     616..635
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(670..1383,1384..1386,1387..1389))
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(715..736)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1064..1083
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1323..1344
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(1411..1437)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(1441..1506)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(1518..1542)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     repeat_region   2200..2329
                     /label=AAV2 ITR
     rep_origin      2401..2856
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2488..2507)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2698..2719
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2882..2986
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2987..3055,3056..3847)
                     /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(3205..3224)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4018..4595,1..11)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4507..4526
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
       61 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
      121 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
      181 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
      241 aatgcagctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc
      301 gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc
      361 catcactagg ggttccttgt agttaatgat taacccgcca tgctacttat ctacgtagcc
      421 atgctctcga ttttaccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacatc
      481 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
      541 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
      601 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgctcga
      661 agcggccgct tacttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa
      721 ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcaggg cggactgggt
      781 gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg
      841 gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg tggcgggtct tgaagttcac
      901 cttgatgccg ttcttctgct tgtcggcggt gatatagacc ttgtggctgt tgtagttgta
      961 ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat
     1021 gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc
     1081 gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa
     1141 gtcgtgctgc ttcatgtggt cggggtagcg ggcgaagcac tgcacgccgt aggtgaaggt
     1201 ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatga acttcagggt
     1261 cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt
     1321 tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt
     1381 gctcaccatt ccgccagcgc ctccacgcgt agcgtaatct ggaacatcgt atgggtaaag
     1441 cttgtcatcg tcatccttgt aatcgatgtc atgatcttta taatcaccgt catggtcttt
     1501 gtagtccatg gttttaaagc ctgctttttt gtacaaactt gttgatagct taactagcca
     1561 gcttgggagg atccagatcc ttggatattt tctagagggt ctggaattcc tgccgacaga
     1621 tcctggaggc gtctgctcag tctcagcggg gactgggtga ggcagaggat ggagagggct
     1681 ttaagcaggc atgtgggctg gggcctggtg agccagccct gcggagggag gaatgtgcga
     1741 caggggacgg gtggggcagg gggatggcgg tgggggtggg gggtgttggc tgctattttg
     1801 gcaggtgcca gggacaaggc tacaggaaca tgtaccccac gccatataag cccatgtggt
     1861 cctccagctg ctcagataag ctatttaaaa ccagagcaga tatgcaggga acagtcatgc
     1921 aacataaacc agctgtccct cttgagaatc ctgataaagc agaggccagc aacccaggcc
     1981 tgggagggcc agctgggagc agggttgggg ggcagaaggc aacctccaag acactccata
     2041 agtctcagca ccagaatctt ggaaggcaga gggcaagagt tatgtgctgc tccacttgaa
     2101 ctgatgctgg gggtaaagac atcttccagg ctactggctc ctaatggact gagcttaagg
     2161 atgtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt
     2221 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     2281 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tggcgtaata
     2341 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg
     2401 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2461 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2521 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     2581 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     2641 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     2701 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     2761 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     2821 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
     2881 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     2941 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     3001 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     3061 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     3121 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     3181 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     3241 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     3301 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     3361 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     3421 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     3481 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     3541 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     3601 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     3661 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     3721 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     3781 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     3841 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     3901 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     3961 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4021 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4081 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4141 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4201 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4261 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4321 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4381 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4441 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4501 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4561 gcgtcgattt ttgtgatgct cgtcaggggg gcgga
//
