LOCUS       sequence-326329-        4984 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-based delivery of GFP into cardiomyocytes in vivo.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4984)
  AUTHORS   Guo Y, Cao Y, Jardin BD, Sethi I, Ma Q, Moghadaszadeh B, Troiano EC,
            Mazumdar N, Trembley MA, Small EM, Yuan GC, Beggs AH, Pu WT
  TITLE     Sarcomeres regulate murine cardiomyocyte maturation through MRTF-SRF
            signaling.
  JOURNAL   Proc Natl Acad Sci U S A. 2021 Jan 12;118(2). pii: 2008861118. doi: 
            10.1073/pnas.2008861118.
  PUBMED    33361330
REFERENCE   2  (bases 1 to 4984)
  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"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2021-01-12"; volume: "118(2). pii: 
            2008861118. doi"; pages: " 10.1073/pnas.2008861118"
COMMENT     Sequence Label: aav-tnt-gfp-v2
FEATURES             Location/Qualifiers
     source          1..4984
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     intron          772..904
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     887..906
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        949..967
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1022..1031
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1028..1030,1031..1033,1034..1747)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1028..1030,1031..1033,1034..1744)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1073..1094)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1334..1353)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1681..1702
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1821..1840)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1886..1941
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   2034..2163
                     /label=AAV2 ITR
     primer_bind     complement(2182..2199)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2182..2198)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2191..2213)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2340..2795
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2427..2446)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2637..2658
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2821..2925
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2926..2994,2995..3786)
                     /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(3144..3163)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3957..4545
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4446..4465
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    4833..4854
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4869..4874,4875..4892,4893..4899)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4907..4923
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     4912..4934
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4931..4947
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4931..4947
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctaccag ggtaatgggg
      181 atcctctaga actatagcta gaattcgccc ttacgggccc cccctcgagg tcgggataaa
      241 agcagtctgg gctttcacat gacagcatct ggggctgcgg cagagggtcg ggtccgaagc
      301 gctgccttat cagcgtcccc agccctggga ggtgacagct ggctggcttg tgtcagcccc
      361 tcgggcactc acgtatctcc gtccgacggg tttaaaatag caaaactctg aggccacaca
      421 atagcttggg cttatatggg ctcctgtggg ggaaggggga gcacggaggg ggccggggcc
      481 gctgctgcca aaatagcagc tcacaagtgt tgcattcctc tctgggcgcc gggcacattc
      541 ctgctggctc tgcccgcccc ggggtgggcg ccggggggac cttaaagcct ctgcccccca
      601 aggagccctt cccagacagc cgccggcacc caccgctccg tgggacgatc cccgaagctc
      661 tagagcttta ttgcggtagt ttatcacagt taaattgcta acgcagtcag tgcttctgac
      721 acaacagtct cgaacttaag ctgcagaagt tggtcgtgag gcactgggca ggtaagtatc
      781 aaggttacaa gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga
      841 ctcttgcgtt tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc
      901 acaggtgtcc actcccagtt caattacagc tcttaaggct agagtactta atacgactca
      961 ctataggcta gcctcgagaa gcggccgcac tactccgcgg actactacta gtttcacgcg
     1021 ggccgccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     1081 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     1141 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     1201 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     1261 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     1321 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     1381 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1441 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     1501 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     1561 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1621 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     1681 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1741 caagtaataa gctcgcgtgg tacctctaga gtcgacccgg gcggcctcga ggacggggtg
     1801 aactacgcct gaggatccga tctttttccc tctgccaaaa attatgggga catcatgaag
     1861 ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt
     1921 tggaattttt tgtgtctctc actcggaagc aattcgttga tctgaatttc gaccacccat
     1981 aatacccatt accctggtag ataagtagca tggcgggtta atcattaact acaaggaacc
     2041 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     2101 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     2161 cagccttaat taacctaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     2221 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     2281 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     2341 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2401 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2461 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2521 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     2581 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2641 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     2701 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     2761 cgcgaatttt aacaaaatat taacgcttac aatttagggg gcacttttcg gggaaatgtg
     2821 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2881 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     2941 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     3001 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     3061 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     3121 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     3181 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     3241 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     3301 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     3361 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     3421 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     3481 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     3541 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     3601 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     3661 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     3721 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     3781 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     3841 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     3901 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3961 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4021 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4081 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     4141 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4201 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4261 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4321 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4381 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4441 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4501 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4561 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     4621 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     4681 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     4741 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     4801 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     4861 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     4921 caatttcaca caggaaacag ctatgaccat gattacgcca gatttaatta aggccttaat
     4981 tagg
//
