LOCUS       sequence-442282-        8170 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mouse fibroblast-to-endothelial cell direct reprogramming.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8170)
  AUTHORS   Farber G, Dong Y, Wang Q, Rathod M, Wang H, Dixit M, Keepers B, Xie 
            Y, Butz K, Polacheck WJ, Liu J, Qian L
  TITLE     Direct conversion of cardiac fibroblasts into endothelial-like cells
            using Sox17 and Erg.
  JOURNAL   Nat Commun. 2024 May 16;15(1):4170. doi: 10.1038/s41467-024-48354-6.
  PUBMED    38755186
REFERENCE   2  (bases 1 to 8170)
  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: "Nat 
            Commun."; date: "2024-05-16"; volume: "15(1):4170. doi"; pages: " 
            10.1038/s41467-024-48354-6"
COMMENT     Sequence Label: pMX-Sox17-p2A-Erg-t2A-GFP
FEATURES             Location/Qualifiers
     source          1..8170
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             22..615
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    678..1035
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1094..1510
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1440..1462
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1478..1494
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1520..1894
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1832..1848
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             3189..3245
                     /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             4725..4778
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(4785..4787,4788..4790,4791..5504)
                     /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(4785..4787,4788..4790,4791..5501)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4830..4851)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5091..5110)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5438..5459
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             5675..6144
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6274..6291)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6445..7033)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6525..6544)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7197..7988,7989..8057))
                     /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     7820..7839
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8058..8162)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cccgaaaagt gccacctgca taatgaaaga ccccacctgt aggtttggca agctagctta
       61 agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaaa agttcagatc
      121 aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
      181 ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat
      241 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
      301 gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
      361 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
      421 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
      481 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
      541 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
      601 cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca gggaccaccg
      661 acccaccacc gggaggtaag ctggccagca acttatctgt gtctgtccga ttgtctagtg
      721 tctatgactg attttatgcg cctgcgtcgg tactagttag ctaactagct ctgtatctgg
      781 cggacccgtg gtggaactga cgagttcgga acacccggcc gcaaccctgg gagacgtccc
      841 agggacttcg ggggccgttt ttgtggcccg acctgagtcc aaaaatcccg atcgttttgg
      901 actctttggt gcacccccct aataggaggg atatgtggtt ctggtaggag acgagaacct
      961 aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tgggaccgaa gccgcgccgc
     1021 gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt
     1081 gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa
     1141 agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac
     1201 cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa
     1261 ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc
     1321 agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt
     1381 caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc
     1441 ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc
     1501 tctaggcgcc cccatatggc catatgagat cttatatggg gcacccccgc cccttgtaaa
     1561 cttccctgac cctgacatga caagagttac taacagcccc tctctccaag ctcacttaca
     1621 ggctctctac ttagtccagc acgaagtctg gagacctctg gcggcagcct accaagaaca
     1681 actggaccga ccggtggtac ctcaccctta ccgagtcggc gacacagtgt gggtccgccg
     1741 acaccagact aagaacctag aacctcgctg gaaaggacct tacacagtcc tgctgaccac
     1801 ccccaccgcc ctcaaagtag acggcatcgc agcttggata cacgccgccc acgtgaaggc
     1861 tgccgacccc gggggtggac catcctctag actgccggat ctagctagtt aattaaatga
     1921 gcagcccgga tgcgggatac gccagtgacg accagagcca gccccggagc gcgcagcccg
     1981 cggtgatggc agggttgggc ccctgtccct gggccgagtc cctgagcccc ctcggggatg
     2041 taaaggtgaa aggcgaggtg gtggcgagta gcggggcgcc agccgggacg tcgggccgag
     2101 ccaaagcgga gtctcgcatc cggcggccga tgaacgcctt tatggtgtgg gccaaagacg
     2161 aacgcaagcg gttggcacag cagaacccag atctgcacaa cgcagagcta agcaagatgc
     2221 taggcaagtc ttggaaggcg ttgaccttgg cagagaagcg gcccttcgtg gaagaggccg
     2281 agcggctgcg cgtgcagcat atgcaggacc accccaacta caagtaccgg ccgcggcggc
     2341 gcaagcaggt gaagcgcatg aagcgggtgg agggaggctt cctgcacgct ctcgtcgagc
     2401 cccaggccgg cgcgcttggt cccgagggcg gccgcgtggc catggatggc ctgggtctgc
     2461 ctttcccgga gccgggctat ccggccggtc ctccgctgat gtctccgcac atgggccccc
     2521 actatcggga ctgccaggga ctgggcgctc ccgcgctcga cggctaccct ctgcccactc
     2581 cggacacatc cccgctggat ggcgtggagc aggacccggc tttctttgca gccccgctgc
     2641 caggggactg cccggcggcc ggcacctaca cttacgctcc agtctcggac tatgcagtgt
     2701 ccgtagagcc gcccgctggc cccatgcgag tggggccgga cccctcgggc cctgcgatgc
     2761 cggggatcct ggcgcccccc agcgctctgc acctgtacta cggcgcgatg ggctcgcccg
     2821 ccgcaagtgc ggggcgcggt ttccacgcgc aaccccagca gccgctgcaa ccgcaggcac
     2881 cgccgccgcc accgcagcag cagcacccag cgcacggccc cgggcaacct tcgccccctc
     2941 ccgaggctct gccctgccgg gatggcacgg aatccaacca gcccactgag ctcctagggg
     3001 aggtggaccg cacggaattc gaacagtatc tgccctttgt gtataagccc gagatgggtc
     3061 ttccctacca gggacacgac tgcggagtga acctctcaga cagccacgga gccatttcct
     3121 ccgtggtgtc cgacgctagc tcagcggtct actattgcaa ctaccccgac attactagcg
     3181 gcagcggcgc cacaaacttc tctctgctaa agcaagcagg tgatgttgaa gaaaaccccg
     3241 ggcctactag tatgatccag actgtacctg acccagcagc tcatattaag gaggccttgt
     3301 cagttgtgag cgaggaccag tcactatttg agtgtgccta cggaacgcca cacctggcta
     3361 agacagagat gaccgcatcc tcttccagtg actatggcca gacatccaag atgagtccca
     3421 gagtccctca gcaggactgg ctgtctcaag ccccagccag ggtcaccatc aagatggagt
     3481 gcaaccctag tcaggtgaat ggttccagga actcacctga tgagtgcagt gtgaacaaag
     3541 gtgggaagat ggtgggcagc ccggatactg tggggatgag ctacggcagc tacatggagg
     3601 agaagcatgt gccgcctccc aatatgacca caaatgagcg cagagtgatc gtccctgcag
     3661 atcctactct gtggagcaca gaccatgtcc gacagtggct ggagtgggcg gtgaaagaat
     3721 atggcctcct cgatgtggac gtcttactat ttcagaatat cgatgggaag gagctgtgca
     3781 agatgacaaa ggatgacttc cagcggctca cgccgagcta caatgccgac attcttctct
     3841 cacatctcca ctacctcaga gagactcccc ttccacatct gacttccgat gacgttgata
     3901 aggctttaca aaactctcca cggttaatgc atgccagaaa cacagggggt gcagctttta
     3961 ttttcccaaa tacttcagta tatcccgaag ctacgcaaag aattacaact aggccagatt
     4021 taccttatga gcctcccagg agatcagcct ggaccggcca cagccacctc acccctcagt
     4081 ccaaagctgc tcagccatct ccctctgcag tgcccaaaac tgaagaccag cgtcctcagt
     4141 tagatcctta ccagatcctg ggaccgacca gtagccgcct tgctaatcca ggtagtggcc
     4201 agatccagct gtggcagttc ctgctcgaac tcctgtcaga cagctccaac tccaactgca
     4261 tcacctggga aggcaccaac ggggagttca agatgacaga cccggacgag gtggctcggc
     4321 gctgggggga gaggaagagc aagcccaaca tgaactatga caagctcagc cgcgccctcc
     4381 gctactacta cgacaaaaac atcatgacca aggtgcacgg gaagcgctac gcctacaagt
     4441 ttgacttcca cgggattgcc caggccctgc agccccaccc tcctgagtcg tccctgtaca
     4501 agtacccctc cgacctgcca tacatgggct cctatcacgc ccacccccag aagatgaact
     4561 ttgtgtctcc ccaccctccc gctctcccag tcacatcttc cagtttcttt gcttccccga
     4621 acccatactg gaattcaccg actgggggca tctacccgaa cactaggctc ccagccagcc
     4681 atatgccctc tcacctgggc acctactaca agcttggctc cggcgagggc aggggaagtc
     4741 ttctaacatg cggggacgtg gaggaaaatc ccggcccact cgagatggtg agcaagggcg
     4801 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     4861 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     4921 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     4981 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     5041 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     5101 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     5161 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     5221 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     5281 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     5341 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     5401 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     5461 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagtcgac gataaaataa
     5521 aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc
     5581 aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact gagaatagag
     5641 aagttcagat caaggtcagg aacagatgga acagctgaat atgggccaaa caggatatct
     5701 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggaacagc tgaatatggg
     5761 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt
     5821 ccccagatgc ggtccagccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc
     5881 cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc
     5941 ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg
     6001 ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt
     6061 gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga
     6121 ctacccgtca gcgggggtct ttcacatgca gcatgtatca aaattaattt ggtttttttt
     6181 cttaagtatt tacattaaat ggccatagtt gcattaatga atcggccaac gcgcggggag
     6241 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6301 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6361 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6421 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6481 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6541 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6601 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6661 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6721 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6781 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6841 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6901 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6961 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7021 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7081 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7141 tttgcggccg gccgcaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7201 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7261 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7321 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7381 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7441 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7501 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7561 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7621 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7681 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7741 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7801 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7861 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7921 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7981 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     8041 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     8101 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     8161 cgcacatttc
//
