LOCUS       sequence-243812-        7712 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7712)
  AUTHORS   Sekiya S, Suzuki A
  TITLE     Direct conversion of mouse fibroblasts to hepatocyte-like cells by 
            defined factors.
  JOURNAL   Nature. 2011 Jun 29;475(7356):390-3. doi: 10.1038/nature10263.
  PUBMED    21716291
REFERENCE   2  (bases 1 to 7712)
  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"; doi: "10"; journalName: 
            "Nature"; date: "2011-06-29- 29"; volume: "475"; issue: "7356"; 
            pages: "390-3"
COMMENT     Sequence Label: pGCDNsam-Hnf4a-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..7712
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             574..749
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    813..1154
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1221..1637
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1567..1589
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1605..1621
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     1906..1922
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     misc_feature    3478..4044
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3494..4046
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3638..3655)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3865..3884
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4058..4060,4061..4063,4064..4777)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4103..4124)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4364..4383)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4711..4732
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4850..5364
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5526..5542
                     /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(5550..5556,5557..5574,5575..5580))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5595..5616
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5733..5750)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5904..6492)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5984..6003)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6663..7454,7455..7523))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7286..7305
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7524..7628)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(7696..7712,1..2)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gccctttcgt cttcaagaat tcataccaga tcaccgaaaa ctgtcctcca aatgtgtccc
       61 cctcacactc ccaaattcgc gggcttctgc ctcttagacc actctaccct attccccaca
      121 ctcaccggag ccaaagccgc ggcccttccg tttctttgct tttgaaagac cccacccgta
      181 ggtggcaagc tagcttaagt aacgccactt tgcaaggcat ggaaaaatac ataactgaga
      241 atagaaaagt tcagatcaag gtcaggaaca aagaaacagc tgaataccaa acaggatatc
      301 tgtggtaagc ggttcctgcc ccggctcagg gccaagaaca gatgagacag ctgagtgatg
      361 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcggggcca agaacagatg
      421 gtccccagat gcggtccagc cctcagcagt ttctagtgaa tcatcagatg tttccagggt
      481 gccccaagga cctgaaaatg accctgtacc ttatttgaac taaccaatca gttcgcttct
      541 cgcttctgtt cgcgcgcttc cgctctccga gctcaataaa agagcccaca acccctcact
      601 cggcgcgcca gtcttccgat agactgcgtc gcccgggtac ccgtattccc aataaagcct
      661 cttgctgttt gcatccgaat cgtggactcg ctgatccttg ggagggtctc ctcagattga
      721 ttgactgccc acctcggggg tctttcattt ggaggttcca ccgagatttg gagacccctg
      781 cccagggacc accgaccccc ccgccgggag gtaagctggc cagcggtcgt ttcgtgtctg
      841 tctctgtctt tgtgcgtgtt tgtgccggca tctaatgttt gcgcctgcgt ctgtactagt
      901 tagctaacta gctctgtatc tggcggaccc gtggtggaac tgacgagttc tgaacacccg
      961 gccgcaaccc tgggagacgt cccagggact ttgggggccg tttttgtggc ccgacctgag
     1021 gaagggagtc gatgtggaat ccgaccccgt caggatatgt ggttctggta ggagacgaga
     1081 acctaaaaca gttcccgcct ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg
     1141 cgtcttgtct gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc
     1201 tgtatttgtc tgaaaattag ggccagactg ttaccactcc cttaagtttg accttaggtc
     1261 actggaaaga tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt
     1321 gggttacctt ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca
     1381 cctttaaccg agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg
     1441 gacacccaga ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc
     1501 cctgggtcaa gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt
     1561 ctctccccct tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca
     1621 ctccttctct aggcgccgga attcgcggcc gtgacaagag ttactaacag cccctctctc
     1681 caagctcact tacaggctct ctacttagtc cagcacgaag tctggagacc tctggcggca
     1741 gcctaccaag aacaactgga ccgaccggtg gtacctcacc cttaccgagt cggcgacaca
     1801 gtgtgggtcc gccgacacca gactaagaac ctagaacctc gctggaaagg accttacaca
     1861 gtcctgctga ccacccccac cgccctcaaa gtagacggca tcgcagcttg gatacacgcc
     1921 gcccacgtga aggctgccga ccccgggggt ggaccatcct ctagactgac gcggccgcgg
     1981 gaattcgatt gtatgcgact ctctaaaacc cttgccggca tggatatggc cgactacagc
     2041 gctgccctgg acccagccta caccaccctg gagtttgaaa atgtgcaggt gttgaccatg
     2101 ggcaatgaca cgtccccatc tgaaggtgcc aacctcaatt catccaacag cctgggcgtc
     2161 agtgccctgt gcgccatctg tggcgaccgg gccaccggca aacactacgg agcctcgagc
     2221 tgtgacggct gcaaggggtt cttcaggagg agcgtgagga agaaccacat gtactcctgc
     2281 aggtttagcc gacaatgtgt ggtagacaaa gataagagga accagtgtcg ttactgcagg
     2341 cttaagaagt gcttccgggc tggcatgaag aaggaagctg tccaaaatga gcgggaccgg
     2401 atcagcacgc ggaggtcaag ctacgaggac agcagcctgc cctccatcaa cgcgctcctg
     2461 caggcagagg ttctgtccca gcagatcacc tctcccatct ctgggatcaa tggcgacatt
     2521 cgggcaaaga agattgccaa catcacagac gtgtgtgagt ctatgaagga gcagctgctg
     2581 gtcctggtcg agtgggccaa gtacatcccg gccttctgcg aactccttct ggatgaccag
     2641 gtggcgctgc tcagggccca cgccggtgag catctgctgc ttggagccac caagaggtcc
     2701 atggtgttta aggacgtgct gctcctaggc aatgactaca tcgtccctcg gcactgtcca
     2761 gagctagcgg agatgagccg tgtgtccatc cgcatcctcg atgagctggt cctgcccttc
     2821 caagagctgc agattgatga caatgaatat gcctgcctca aagccatcat cttctttgat
     2881 ccagatgcca aggggctgag tgacccgggc aagatcaagc ggctgcggtc acaggtgcaa
     2941 gtgagcctgg aggattacat caacgaccgg cagtacgact ctcggggccg ctttggagag
     3001 ctgctgctgc tgttgcccac gctgcagagc atcacctggc agatgatcga acagatccag
     3061 ttcatcaagc tcttcggcat ggccaagatt gacaacctgc tgcaggagat gcttctcgga
     3121 gggtctgcca gtgatgcacc ccacacccac caccccctgc accctcacct gatgcaagaa
     3181 cacatgggca ccaatgtcat tgttgctaac acgatgccct ctcacctcag caatggacag
     3241 atgtgtgagt ggccccgacc cagggggcag gcagccactc ccgagactcc acagccatca
     3301 ccaccaagtg gctcgggatc tgaatcctac aagctcctgc caggagccat caccaccatc
     3361 gtcaagcctc cctctgccat tccccagcca acgatcacca agcaagaagc catctaggca
     3421 atcactagtg aattcgcggc cgctacgtag tcgacggatc cctcgaggtt aacgaattcc
     3481 gccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc
     3541 tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc
     3601 cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt
     3661 ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct
     3721 gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa
     3781 aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt
     3841 tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag
     3901 gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt
     3961 acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt
     4021 ttcctttgaa aaacacgatg ataatatggc cacaaccatg gtgagcaagg gcgaggagct
     4081 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     4141 cagcgtgtct ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     4201 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     4261 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     4321 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     4381 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     4441 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     4501 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggcga acttcaagat
     4561 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     4621 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     4681 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     4741 cgggatcact ctcggcatgg acgagctgta caagtaatga attaattaag aattatcaag
     4801 cttatcgata aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc
     4861 cacctgtagg tttggcaagc tagcttaagt aagcttattt tgcaaggcat ggaaaataca
     4921 taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca gaatatgggc
     4981 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc
     5041 cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc
     5101 ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct
     5161 tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc
     5221 gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt gtatccaata aaccctcttg
     5281 cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac
     5341 tacccgtcag cgggggtctt tcatgggtaa cagtttcttg aagttggaga acaacattct
     5401 gagggtagga gtcgaatatt aagtaatcct gactcaatta gccactgttt tgaatccaca
     5461 tactccaata ctcctgaaat agttcattat ggacagcgca gaaagagctg gggagaattg
     5521 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     5581 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     5641 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     5701 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5761 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5821 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5881 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5941 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6001 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6061 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6121 agttcggtgt aggtcgttcg ctccaagctg ggctatgtgc acgaaccccc cgttcagccc
     6181 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6241 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6301 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     6361 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6421 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6481 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6541 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6601 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6661 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6721 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6781 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6841 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6901 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6961 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7021 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7081 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7141 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7201 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7261 tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg
     7321 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7381 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7441 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7501 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7561 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7621 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     7681 acattaacct ataaaaatag gcgtatcacg ag
//
