LOCUS       Exported                4708 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV vector for Atrial specific EmGFP expression.
ACCESSION   .
VERSION     .
KEYWORDS    013LP-pAAV-ANF-657-EmGFP-pA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4708)
  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 4708)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4708
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   4..133
                     /label=AAV2 ITR
     protein_bind    904..928
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             940..1005
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1009..1035
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1057..1776
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1102..1123)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1363..1382)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1710..1731
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1800..1921
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1811..1830)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1865..1884
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2069..2198
                     /label=AAV2 ITR
     primer_bind     complement(2264..2281)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2435..3023)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2515..2534)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3194..4054)
                     /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     3817..3836
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4055..4159)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4185..4640)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4322..4343)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4534..4553
                     /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 atccttaagg
      181 ggccctgtat catgttggct tcctggctga cttcatactc taaaaaaaaa ataatagctc
      241 tttcacctga ctgctaacag ggacatctag ggtgggggtg ggctgtctgg ggccagaggt
      301 ccacccacga ggccgatgaa tcaggtgtga agctagctcc agcatgtgta ctccctggcc
      361 agcctagctg gcctcccagc tgcctgtcat tgcctcctct cccgccctta tttggagccc
      421 ctgacagctg agcagcaagc agggggagct gggtgcaggc cagccgtcac cctctgcttc
      481 cctgcatggg tcctgttgcc agggagaaag aatcctgagg cgagcgccca ggaagataac
      541 caaggactct tttctgctcc tctcacacct ttgaagtggg ggcctcttga ggcaaatcag
      601 caagaatgtg actcttgcag ctgagggtct gggggagggg ggtgagtgga gctgctcaag
      661 gcaaaggggc cgtgacaagc tttgccgaac tgataacttt aaaagggcat cttctgctgg
      721 ctcctcactc catcgcttta tcgctgcaag tgacagaatg gggagggttc tagcccccct
      781 gccttctcaa agagctgggg ggctataaaa acgggagatg ctggcagcta ggagacgaat
      841 tccagaccct ctagaaaata tccaaggatc tggatcctcc caagctggct agttaagcta
      901 tcaacaagtt tgtacaaaaa agcaggcttt aaaaccatgg actacaaaga ccatgacggt
      961 gattataaag atcatgacat cgattacaag gatgacgatg acaagcttta cccatacgat
     1021 gttccagatt acgctacgcg tggaggcgct ggcggaatgg tgagcaaggg cgaggagctg
     1081 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     1141 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     1201 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cacctacggc
     1261 gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     1321 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     1381 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     1441 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     1501 cacaaggtct atatcaccgc cgacaagcag aagaacggca tcaaggtgaa cttcaagacc
     1561 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     1621 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     1681 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     1741 gggatcactc tcggcatgga cgagctgtac aagtaagcgg ccgcttcgag cagacatgat
     1801 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     1861 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     1921 taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt
     1981 ttaaagcaag taaaacctct acaaatgtgg taaaatcgag agcatggcta cgtagataag
     2041 tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt ggccactccc
     2101 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     2161 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcattaatga atcggccaac
     2221 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     2281 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     2341 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2401 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2461 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2521 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2581 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2641 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2701 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2761 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     2821 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     2881 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     2941 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3001 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3061 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3121 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3181 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3241 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3301 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3361 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3421 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3481 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     3541 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     3601 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     3661 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     3721 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     3781 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     3841 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     3901 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     3961 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4021 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     4081 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4141 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa
     4201 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     4261 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     4321 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     4381 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     4441 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     4501 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     4561 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     4621 tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg
     4681 atcggtgcgg gcctcttcgc tattacgc
//
