LOCUS       sequence-353289-        5030 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dual fluorescent reporter for MYH6 (short) and TNNT2 promoters.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5030)
  AUTHORS   Romero-Tejeda M, Fonoudi H, Weddle CJ, DeKeyser JM, Lenny B, 
            Fetterman KA, Magdy T, Sapkota Y, Epting C, Burridge PW
  TITLE     A novel transcription factor combination for direct reprogramming to
            a spontaneously contracting human cardiomyocyte-like state.
  JOURNAL   J Mol Cell Cardiol. 2023 Jul 6:S0022-2828(23)00106-2. doi: 
            10.1016/j.yjmcc.2023.06.005.
  PUBMED    37421991
REFERENCE   2  (bases 1 to 5030)
  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: "J Mol Cell 
            Cardiol. 2023 Jul 6:S0022-2828(23)00106-2. doi: 
            10.1016/j.yjmcc.2023.06.005."
COMMENT     Sequence Label: pFU-GW-sMYH6-mNeon-TNNT2-mScarlet
FEATURES             Location/Qualifiers
     source          1..5030
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     181..201
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(210..638)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(320..341)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     532..551
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    684..908
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     897..914
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             940..1120
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(1138..1336)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1163..1183)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1338..1717
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1889..1908
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1983..2087
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2088..2156,2157..2948)
                     /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(2306..2325)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3119..3707
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3608..3627
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3861..3878
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3995..4016
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4031..4036,4037..4054,4055..4061)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4069..4085
                     /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     4074..4096
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4093..4109
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4093..4109
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4158..4279
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4169..4188)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4223..4242
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4409..4783)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(4802..4849)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(4897..5030,1..196))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(4911..5030,1..16)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4965..4984)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat
       61 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg
      121 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
      181 accctaactg acacacattc cacagaatta attcgcgtta aatttttgtt aaatcagctc
      241 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      301 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      361 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      421 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      481 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      541 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      601 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtgg ggataccccc tagagcccca
      661 gctggttctt tccgcctcag aagccataga gcccaccgca tccccagcat gcctgctatt
      721 gtcttcccaa tcctccccct tgctgtcctg ccccacccca ccccccagaa tagaatgaca
      781 cctactcaga caatgcgatg caatttcctc attttattag gaaaggacag tgggagtggc
      841 accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct ggcaactaga
      901 aggcacagtc gaggctgatc agcgggttta aacgggccct gctagagatt ttccacactg
      961 actaaaaggg tctgagggat ctctagttac cagagtcaca caacagacgg gcacacacta
     1021 cttgaagcac tcaaggcaag ctttattgag gcttaagcag tgggttccct agttagccag
     1081 agagctccca ggctcagatc tggtctaacc agagagaccc agtacaggca aaacgcgctg
     1141 cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg
     1201 ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca
     1261 atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac
     1321 tgccaaaacc gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta
     1381 ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg
     1441 acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt
     1501 taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga
     1561 acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca
     1621 tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta
     1681 attgattact attaataact agtcaataat caatgtcaac gcgtatatct ggcccgtaca
     1741 tcgcgaagca gcgcaaaacg cctaacccta agcagattct tcatgcaatt gtcggtcaag
     1801 ccttgccttg ttgtagctta aattttgctc gcgcactact cagcgacctc caacacacaa
     1861 gcagggagca gatactggct taactatgcg gcatcagagc agattgtact gagagtgcac
     1921 cataggggat cgggagatct cccgatccgt cgacgtcagg tggcactttt cggggaaatg
     1981 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     2041 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     2101 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     2161 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     2221 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     2281 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     2341 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     2401 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     2461 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     2521 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     2581 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     2641 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     2701 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     2761 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     2821 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     2881 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     2941 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     3001 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     3061 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3121 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3181 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3241 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     3301 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3361 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3421 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3481 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3541 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3601 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3661 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3721 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     3781 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     3841 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     3901 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     3961 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     4021 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     4081 aacaatttca cacaggaaac agctatgacc atgattacgc caagctctag ctagaggtcg
     4141 acggtataca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     4201 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     4261 aagctgcaat aaacaagttg gggtgggcga agaactccag catgagatcc ccgcgctgga
     4321 ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca tagaaggcgg
     4381 cggtggaatc gaaatctcgt agcacgtgtc agtcctgctc ctcggccacg aagtgcacgc
     4441 agttgccggc cgggtcgcgc agggcgaact cccgccccca cggctgctcg ccgatctcgg
     4501 tcatggccgg cccggaggcg tcccggaagt tcgtggacac gacctccgac cactcggcgt
     4561 acagctcgtc caggccgcgc acccacaccc aggccagggt gttgtccggc accacctggt
     4621 cctggaccgc gctgatgaac agggtcacgt cgtcccggac cacaccggcg aagtcgtcct
     4681 ccacgaagtc ccgggagaac ccgagccggt cggtccagaa ctcgaccgct ccggcgacgt
     4741 cgcgcgcggt gagcaccgga acggcactgg tcaacttggc catggtttag ttcctcacct
     4801 tgtcgtatta tactatgccg atatactatg ccgatgatta attgtcaaca cgtgctgatc
     4861 agatccgaaa atggatatac aagctcccgg gagctttttg caaaagccta ggcctccaaa
     4921 aaagcctcct cactacttct ggaatagctc agaggcagag gcggcctcgg cctctgcata
     4981 aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg
//
