LOCUS       sequence-501683-        5853 bp    DNA     circular SYN 04-MAR-2026
DEFINITION  Expresses Cre and TagRFP under the control of the Nppa promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5853)
  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 5853)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 4, 2026 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-Cre-P2A-TagRFP
FEATURES             Location/Qualifiers
     source          1..5853
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             828..848
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1887..1943
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1959..2672
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     primer_bind     complement(2692..2711)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2757..2812
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   2905..3034
                     /label=AAV2 ITR
     primer_bind     complement(3053..3070)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3053..3069)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3062..3084)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3211..3666
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3298..3317)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3508..3529
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3692..3796
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3797..3865,3866..4657)
                     /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(4015..4034)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4828..5416
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5317..5336
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5704..5725
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5740..5745,5746..5763,5764..5770)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5778..5794
                     /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     5783..5805
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5802..5818
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5802..5818
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ggctgcgcgc tcgctcgctc actgaggccg cccgggcaaa gcccgggcgg cctcagtgag
       61 cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct tgtagttaat
      121 gattaacccg ccatgctact tatctaccag ggtaatgggg atccgggccc tgtatcatgt
      181 tggcttcctg gctgacttca tactctaaaa aaaaaataat agctctttca cctgactgct
      241 aacagggaca tctagggtgg gggtgggctg tctggggcca gaggtccacc cacgaggccg
      301 atgaatcagg tgtgaagcta gctccagcat gtgtactccc tggccagcct agctggcctc
      361 ccagctgcct gtcattgcct cctctcccgc ccttatttgg agcccctgac agctgagcag
      421 caagcagggg gagctgggtg caggccagcc gtcaccctct gcttccctgc atgggtcctg
      481 ttgccaggga gaaagaatcc tgaggcgagc gcccaggaag ataaccaagg actcttttct
      541 gctcctctca cacctttgaa gtgggggcct cttgaggcaa atcagcaaga atgtgactct
      601 tgcagctgag ggtctggggg aggggggtga gtggagctgc tcaaggcaaa ggggccgtga
      661 caagctttgc cgaactgata actttaaaag ggcatcttct gctggctcct cactccatcg
      721 ctttatcgct gcaagtgaca gaatggggag ggttctagcc cccctgcctt ctcaaagagc
      781 tggggggcta taaaaacggg agatgctggc agctaggaga caccatgccc aagaagaaga
      841 ggaaggtgcc atggtctaga tccaatctcc tgactgttca ccagaacctc cctgcgctgc
      901 cagtagatgc cactagcgat gaggtcagga aaaatctcat ggatatgttt agggatagac
      961 aggcgttttc tgaacacacc tggaaaatgc tgcttagcgt gtgccgatcc tgggcagcct
     1021 ggtgtaagct gaacaatcgc aaatggttcc ccgccgagcc ggaggacgtg cgcgattacc
     1081 tgctgtatct ccaggcaaga gggctggctg tcaagactat ccagcagcac ttgggccaac
     1141 tgaatatgct gcatcgacgc agcgggctcc cccggcctag cgattcaaac gcagtctccc
     1201 ttgttatgag gagaattaga aaggaaaacg tagatgcggg tgagagggct aagcaggctc
     1261 tcgcttttga gcggactgat ttcgaccagg tcagatccct gatggagaac agcgatcggt
     1321 gccaggacat caggaacctc gcatttctgg gaattgcata taacacactt ctgcgcatag
     1381 ctgagatcgc ccggatcaga gtgaaagaca tcagtcgaac ggacggcggc cggatgctta
     1441 ttcatattgg acgcacaaag acattggtca gcaccgctgg cgttgaaaag gccttgtccc
     1501 tgggcgtaac gaagctggtg gaaagatgga tctcagtgtc cggcgtggct gacgacccta
     1561 ataattactt gttctgtcga gtgagaaaaa acggagtcgc cgcgccctct gccaccagcc
     1621 aattgagtac acgggccctt gaagggatct ttgaggcaac ccaccgactc atatacggag
     1681 ccaaggatga cagtggccag aggtatctcg cctggtcagg tcattctgct agggtggggg
     1741 ccgcacgaga catggcgcgg gcaggagtct ccataccaga gattatgcaa gctggaggtt
     1801 ggacaaatgt gaacatcgtt atgaactata tccgcaatct tgactctgaa accggggcca
     1861 tggtgagact gctcgaagat ggtgacgcca ccaacttctc cctgctgaag caggccggcg
     1921 acgtggagga gaaccccggc cccctcgagg gcggccgcat ggtgtctaag ggcgaagagc
     1981 tgattaagga gaacatgcac atgaagctgt acatggaggg caccgtgaac aaccaccact
     2041 tcaagtgcac atccgagggc gaaggcaagc cctacgaggg cacccagacc atgagaatca
     2101 aggtggtcga gggcggccct ctccccttcg ccttcgacat cctggctacc agcttcatgt
     2161 acggcagcag aaccttcatc aaccacaccc agggcatccc cgacttcttt aagcagtcct
     2221 tccctgaggg cttcacatgg gagagagtca ccacatacga agacgggggc gtgctgaccg
     2281 ctacccagga caccagcctc caggacggct gcctcatcta caacgtcaag atcagagggg
     2341 tgaacttccc atccaacggc cctgtgatgc agaagaaaac actcggctgg gaggccaaca
     2401 ccgagatgct gtaccccgct gacggcggcc tggaaggcag aagcgacatg gccctgaagc
     2461 tcgtgggcgg gggccacctg atctgcaact tcaagaccac atacagatcc aagaaacccg
     2521 ctaagaacct caagatgccc ggcgtctact atgtggacca cagactggaa agaatcaagg
     2581 aggccgacaa agagacctac gtcgagcagc acgaggtggc tgtggccaga tactgcgacc
     2641 tccctagcaa actggggcac aaacttaatt gaggtaccgg atccgatccg atctttttcc
     2701 ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata
     2761 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag
     2821 caattcgttg atctgaattt cgaccaccca taatacccat taccctggta gataagtagc
     2881 atggcgggtt aatcattaac tacaaggaac ccctagtgat ggagttggcc actccctctc
     2941 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     3001 cccgggcggc ctcagtgagc gagcgagcgc gcagccttaa ttaacctaat tcactggccg
     3061 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     3121 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     3181 aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
     3241 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3301 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3361 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3421 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3481 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3541 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     3601 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     3661 caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3721 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3781 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3841 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3901 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3961 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4021 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4081 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     4141 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4201 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4261 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     4321 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     4381 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     4441 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4501 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4561 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4621 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     4681 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4741 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4801 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4861 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4921 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     4981 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5041 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5101 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5161 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5221 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5281 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5341 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5401 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5461 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     5521 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     5581 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     5641 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     5701 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     5761 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     5821 tgattacgcc agatttaatt aaggccttaa tta
//
