LOCUS       sequence-491534-        4840 bp    DNA     circular SYN 04-NOV-2025
DEFINITION  Expresses EGFP under the control of the Nppa promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4840)
  AUTHORS   Sweat ME, Cao Y, Zhang X, Burnicka-Turek O, Perez-Cervantes C, 
            Arulsamy K, Lu F, Keating EM, Akerberg BN, Ma Q, Wakimoto H, Gorham 
            JM, Hill LD, Kyoung Song M, Trembley MA, Wang P, Gianeselli M, 
            Prondzynski M, Bortolin RH, Bezzerides VJ, Chen K, Seidman JG, 
            Seidman CE, Moskowitz IP, Pu WT
  TITLE     Tbx5 maintains atrial identity in post-natal cardiomyocytes by 
            regulating an atrial-specific enhancer network.
  JOURNAL   Nat Cardiovasc Res. 2023 Oct;2(10):881-898. doi: 
            10.1038/s44161-023-00334-7. Epub 2023 Oct 12.
  PUBMED    38344303
REFERENCE   2  (bases 1 to 4840)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 4, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s44161-023-00334-7"; journalName: "Nat Cardiovasc Res"; 
            date: "2023-10"; volume: "2"; issue: "10"; pages: "881-898"
COMMENT     Sequence Label: AAV-Nppa-EGFP
FEATURES             Location/Qualifiers
     source          1..4840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(645..661)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(645..661)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(658..680)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    669..685
                     /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)."
     promoter        complement(join(693..699,700..717,718..723))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    738..759
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(1047..1635)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1127..1146)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1806..2597,2598..2666))
                     /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     2429..2448
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2667..2771)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2797..3252)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2934..2955)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3146..3165
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3379..3401
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3393..3410
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3394..3410
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   3429..3558
                     /label=AAV2 ITR
     polyA_signal    3651..3706
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     3752..3771
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     misc_feature    3801..3866
                     /label=MCS
                     /note="multiple cloning site"
     CDS             complement(join(3867..4577,4578..4580,4581..4583))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(3867..4577,4578..4580,4581..4583))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3909..3930)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4258..4277
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4517..4538
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(4602..4622)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 tcagctgcaa gagtcacatt cttgctgatt tgcctcaaga ggcccccact tcaaaggtgt
       61 gagaggagca gaaaagagtc cttggttatc ttcctgggcg ctcgcctcag gattctttct
      121 ccctggcaac aggacccatg cagggaagca gagggtgacg gctggcctgc acccagctcc
      181 ccctgcttgc tgctcagctg tcaggggctc caaataaggg cgggagagga ggcaatgaca
      241 ggcagctggg aggccagcta ggctggccag ggagtacaca tgctggagct agcttcacac
      301 ctgattcatc ggcctcgtgg gtggacctct ggccccagac agcccacccc caccctagat
      361 gtccctgtta gcagtcaggt gaaagagcta ttattttttt tttagagtat gaagtcagcc
      421 aggaagccaa catgatacag ggcccggatc cccattaccc tggtagataa gtagcatggc
      481 gggttaatca ttaactacaa ggaaccccta gtgatggagt tggccactcc ctctctgcgc
      541 gctcgctcgc tcactgaggc cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
      601 cgcgcagcct aattaaggcc ttaattaaat ctggcgtaat catggtcata gctgtttcct
      661 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
      721 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
      781 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
      841 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
      901 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
      961 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     1021 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     1081 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     1141 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     1201 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     1261 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     1321 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     1381 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     1441 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     1501 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     1561 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     1621 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     1681 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     1741 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     1801 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     1861 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     1921 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     1981 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     2041 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     2101 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     2161 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     2221 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     2281 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     2341 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     2401 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     2461 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     2521 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     2581 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     2641 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     2701 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     2761 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt
     2821 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     2881 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     2941 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     3001 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     3061 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     3121 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     3181 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     3241 tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     3301 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     3361 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattaggtt
     3421 aattaaggct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg
     3481 cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact
     3541 ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta tctaccaggg
     3601 taatgggtat tatgggtggt cgaaattcag atcaacgaat tgcttccgag tgagagacac
     3661 aaaaaattcc aacacactat tgcaatgaaa ataaatttcc tttattagcc agaagtcaga
     3721 tgctcaaggg gcttcatgat gtccccataa tttttggcag agggaaaaag atcggatcgg
     3781 gtaccttatc tagatccggt ggatcccggg cccgcggtac cgtcgactgc agaattcgaa
     3841 gcttgagctc gagatctgag tccggacttg tacagctcgt ccatgccgag agtgatcccg
     3901 gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc
     3961 agggcggact gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg
     4021 ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg
     4081 atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg
     4141 ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
     4201 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
     4261 ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg
     4321 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg
     4381 ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag
     4441 atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg
     4501 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
     4561 agctcctcgc ccttgctcac catgaattct ctagaccatg gcaccttcct cttcttcttg
     4621 ggcatggtgt ctcctagctg ccagcatctc ccgtttttat agccccccag ctctttgaga
     4681 aggcaggggg gctagaaccc tccccattct gtcacttgca gcgataaagc gatggagtga
     4741 ggagccagca gaagatgccc ttttaaagtt atcagttcgg caaagcttgt cacggcccct
     4801 ttgccttgag cagctccact cacccccctc ccccagaccc
//
