LOCUS       sequence-423016-        7012 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cardiomyocytes-specific plasmid, expressing the coding sequence of 
            mouse Nexn and GFP tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7012)
  AUTHORS   Guo CT, Jardin BD, Lin JS, Ambroise RL, Wang Z, Yang LZ, Mazumdar N,
            Lu FJ, Ma Q, Cao YP, Liu CZ, Li KL, Liu XJ, Lan F, Zhao MM, Xiao H, 
            Dong ED, Pu WT, Guo YX
  TITLE     In vivo proximity proteomics uncovers palmdelphin (PALMD) as a 
            Z-disc-associated mitigator of isoproterenol-induced cardiac injury.
  JOURNAL   Acta Pharmacol Sin. 2024 Jul 23. doi: 10.1038/s41401-024-01348-y.
  PUBMED    39043970
REFERENCE   2  (bases 1 to 7012)
  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: "Acta 
            Pharmacol Sin. 2024 Jul 23. doi: 10.1038/s41401-024-01348-y."
COMMENT     Sequence Label: pAAV-cTNT-NEXN-GFP
FEATURES             Location/Qualifiers
     source          1..7012
                     /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"
     CDS             join(3056..3058,3059..3061,3062..3775)
                     /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(3056..3058,3059..3061,3062..3772)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3101..3122)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3362..3381)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3709..3730
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3849..3868)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3914..3969
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   4062..4191
                     /label=AAV2 ITR
     primer_bind     complement(4210..4227)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4210..4226)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4219..4241)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4368..4823
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4455..4474)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4665..4686
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4849..4953
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4954..5022,5023..5814)
                     /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(5172..5191)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5985..6573
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6474..6493
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6861..6882
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6897..6902,6903..6920,6921..6927)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6935..6951
                     /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     6940..6962
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6959..6975
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6959..6975
                     /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 gtatgaatga
     1021 cgtttcgcaa aaggcagaga ttctgctttc ctcatgtaaa cctgtcccta agtcctatgt
     1081 gccaaagctt ggcaagggag atgtaaagga caagtttgaa gccatgcaga gggccaggga
     1141 agaaagaaat caaaggcgat ctagagacga aaagcaaaga agaaaagaac aatatattag
     1201 agagagagaa tggaacagga gaaagcagga gattaaagaa atgcttgctt ctgatgatga
     1261 ggaagaaagt tctcctaaaa tagaaaaggc ttatgtccca aagctaacag ggacagttaa
     1321 aggtaaattc gatgaaatgg agaaacacag acaggaagaa caaagaaaga gaacagaaga
     1381 agaaagaaaa cgcagaattg agcaggatct gttagaaaag aggaaaatac agcgcgaatt
     1441 agcaaaaaga gcggagcaga ttgaggacat aaacaatacg ggaaccgagt ccgcatcaga
     1501 ggaaggagat gactcactac ttataacagt ggtacctgcc aaatcataca aaacacctgg
     1561 aaaaacaaag gatcctgagg acctggacag agaagaagga aatgggagaa ccaaccatga
     1621 agaagataaa atgagatatg aagaagagtg ccgagttctc aaagaagcaa aatgtctttc
     1681 attagtcatg gatgatgaaa cagaggccaa aaaagaatca cattttcctg gaaaattgaa
     1741 atcaactttt gaagaactgg aacgacaaag acaagaaaat cgcaagaagc aagcggaaga
     1801 ggaagcgaga cgacgcttag aagaagagag acgttccttt gaggaggcga gaaggcacat
     1861 ggtaaatgag gaggatgaaa accaagatag agaaacagtt tttaaagagt accgcccagg
     1921 aaaactcaaa ctcagttttg aggaaataga aaggcaaagg agagaagacg aaaaacgaaa
     1981 agcagaggag gaggcccgaa ggagaataga ggaagagaag gcggcatttg ctgaagcaag
     2041 gagaagcatg gtattagatg atgattcccc agagatttat aaaacagtgt ctcaagaatc
     2101 tcttactcct ggaaaactag aaattaattt tgaacaatta ttaagacaaa aaatggaaga
     2161 agaaagacga cgaacagagg aagaacgcag gcataaacta gaaatggaga aacaggaatt
     2221 tgaacaactg agacaagaaa tgggaaagga agaagaagaa aatgaaagct ttgggttgag
     2281 tagagaatat gaagaattaa ttaaattaaa aaggagtggc tcaattcaag ctaaaaatct
     2341 aaaaagcaag tttgaaaaaa ttggacaatt gtctgaaaaa gaagtacaga aaaagataga
     2401 agaagagcga gcaaaaaggc gagcaattga ccttgaaatc aaagagcgag aagcagaaaa
     2461 cttccatgag gacgatgatg ttgatgtaag gcctgcaaag aagagtgagt ctccttttac
     2521 tcataaggtg aacatgaaag ccaggtttga acaaatggca aaggcaagag aagaggaaga
     2581 acaaagaaga attgaagaac aaaagttact acgaatgcag tttgaacaga aagaaatcga
     2641 tgcggcatta cagaagaaaa gagaagatga ggaggaagaa gaaggtagca ttgttaatgg
     2701 ctccactact gaagatgaag agcaaaccag atcaggagct ccatggttca aaaagccttt
     2761 aagaaatacc tcagttgtag acagtgagcc agtcagattt actgttaaag taacaggaga
     2821 acccaagccg gaaattacat ggtggtttga aggagaaata ctgcaggatg gagaagacta
     2881 tcagtacatc gaaagaggtg aaacttactg cctgtattta ccggaaacct tcccagaaga
     2941 tggaggagag tacatgtgta aggcagtcaa caataaaggc tcagcagcga gcacctgcat
     3001 tcttaccatt gaaatggatg actacggtgg cggtggctcg ggcggtggtg ggtcgatggt
     3061 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     3121 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     3181 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     3241 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     3301 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     3361 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     3421 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     3481 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     3541 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     3601 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     3661 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     3721 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaataagc
     3781 tcgcgtggta cctctagagt cgacccgggc ggcctcgagg acggggtgaa ctacgcctga
     3841 ggatccgatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat
     3901 ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg
     3961 tgtctctcac tcggaagcaa ttcgttgatc tgaatttcga ccacccataa tacccattac
     4021 cctggtagat aagtagcatg gcgggttaat cattaactac aaggaacccc tagtgatgga
     4081 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     4141 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gccttaatta
     4201 acctaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     4261 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     4321 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag
     4381 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4441 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4501 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4561 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4621 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4681 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4741 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     4801 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
     4861 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4921 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     4981 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5041 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     5101 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     5161 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     5221 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     5281 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5341 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5401 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5461 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5521 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5581 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5641 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5701 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5761 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5821 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5881 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5941 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6001 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6061 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6121 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6181 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6241 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6301 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6361 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6421 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6481 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6541 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6601 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6661 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     6721 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     6781 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     6841 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     6901 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     6961 ggaaacagct atgaccatga ttacgccaga tttaattaag gccttaatta gg
//
