LOCUS       Exported                7682 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pGCDNsam-Foxa1-IRES-GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7682)
  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 7682)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7682
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    814..1366
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(958..975)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1185..1204
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1378..2097
                     /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(1423..1444)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1684..1703)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2031..2052
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    2846..2862
                     /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(2870..2900)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2915..2936
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3053..3070)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3224..3812)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3304..3323)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3983..4843)
                     /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     4606..4625
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4844..4948)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5016..5034
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     LTR             5606..5781
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    5845..6186
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             6253..6669
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6599..6621
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6637..6653
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     6938..6954
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
ORIGIN
        1 actcgctgtc cttcaacgat tgtttcgtca aggtggcacg atccccggac aagccaggca
       61 agggctccta ctggacgctg cacccggact ccggcaacat gttcgagaac ggctgctact
      121 tgcgccgcca aaagcgcttc aagtgtgaga agcagccggg ggccggaggt gggagtgggg
      181 gcggcggctc caaagggggc ccagaaagtc gcaaggaccc ctcaggcccg gggaacccca
      241 gcgccgagtc accccttcac cggggtgtgc acggaaaggc tagccagcta gagggcgcgc
      301 cggccccagg gcccgccgcc agcccccaga ctctggacca cagcggggcc acggcgacag
      361 ggggcgcttc ggagttgaag tctccagcgt cttcatctgc gccccccata agctccgggc
      421 caggggcgct agcatctgta cccccctctc acccggctca cggcctggca ccccacgaat
      481 ctcagctgca tctgaaaggg gatccccact actcctttaa tcaccccttc tccatcaaca
      541 acctcatgtc ctcctccgag caacagcaca agctggactt caaggcatac gagcaggcgc
      601 tgcagtactc tccttatggc gctaccttgc ccgccagtct gccccttggc agcgcctcag
      661 tggccacgag gagccccatc gagccctcag ccctggagcc agcctactac caaggtgtgt
      721 attccagacc cgtgctaaat acttcctaga caatcactag tgaattgtag tcgacggatc
      781 cctcgaggtt aacgaattcc gccccccccc ctaacgttac tggccgaagc cgcttggaat
      841 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
      901 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
      961 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     1021 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     1081 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     1141 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     1201 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     1261 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     1321 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg
     1381 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     1441 gacgtaaacg gccacaagtt cagcgtgtct ggcgagggcg agggcgatgc cacctacggc
     1501 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     1561 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     1621 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     1681 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     1741 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     1801 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     1861 atcaaggcga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     1921 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1981 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     2041 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaatga
     2101 attaattaag aattatcaag cttatcgata aaataaaaga ttttatttag tctccagaaa
     2161 aaggggggaa tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aagcttattt
     2221 tgcaaggcat ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag
     2281 agagacagca gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc
     2341 agggccaaga acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca
     2401 tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac
     2461 caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag
     2521 cccacaaccc ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt
     2581 gtatccaata aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg
     2641 gtctcctctg agtgattgac tacccgtcag cgggggtctt tcatgggtaa cagtttcttg
     2701 aagttggaga acaacattct gagggtagga gtcgaatatt aagtaatcct gactcaatta
     2761 gccactgttt tgaatccaca tactccaata ctcctgaaat agttcattat ggacagcgca
     2821 gaaagagctg gggagaattg tgaaattgtt atccgctcac aattccacac aacatacgag
     2881 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     2941 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     3001 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3061 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3121 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3181 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3241 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3301 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3361 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3421 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3481 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3541 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3601 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3661 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3721 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3781 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     3841 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     3901 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     3961 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4021 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4081 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4141 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4201 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4261 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4321 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4381 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     4441 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4501 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4561 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac
     4621 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     4681 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     4741 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     4801 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     4861 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     4921 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     4981 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     5041 gtcttcaaga attcatacca gatcaccgaa aactgtcctc caaatgtgtc cccctcacac
     5101 tcccaaattc gcgggcttct gcctcttaga ccactctacc ctattcccca cactcaccgg
     5161 agccaaagcc gcggcccttc cgtttctttg cttttgaaag accccacccg taggtggcaa
     5221 gctagcttaa gtaacgccac tttgcaaggc atggaaaaat acataactga gaatagaaaa
     5281 gttcagatca aggtcaggaa caaagaaaca gctgaatacc aaacaggata tctgtggtaa
     5341 gcggttcctg ccccggctca gggccaagaa cagatgagac agctgagtga tgggccaaac
     5401 aggatatctg tggtaagcag ttcctgcccc ggctcggggc caagaacaga tggtccccag
     5461 atgcggtcca gccctcagca gtttctagtg aatcatcaga tgtttccagg gtgccccaag
     5521 gacctgaaaa tgaccctgta ccttatttga actaaccaat cagttcgctt ctcgcttctg
     5581 ttcgcgcgct tccgctctcc gagctcaata aaagagccca caacccctca ctcggcgcgc
     5641 cagtcttccg atagactgcg tcgcccgggt acccgtattc ccaataaagc ctcttgctgt
     5701 ttgcatccga atcgtggact cgctgatcct tgggagggtc tcctcagatt gattgactgc
     5761 ccacctcggg ggtctttcat ttggaggttc caccgagatt tggagacccc tgcccaggga
     5821 ccaccgaccc ccccgccggg aggtaagctg gccagcggtc gtttcgtgtc tgtctctgtc
     5881 tttgtgcgtg tttgtgccgg catctaatgt ttgcgcctgc gtctgtacta gttagctaac
     5941 tagctctgta tctggcggac ccgtggtgga actgacgagt tctgaacacc cggccgcaac
     6001 cctgggagac gtcccaggga ctttgggggc cgtttttgtg gcccgacctg aggaagggag
     6061 tcgatgtgga atccgacccc gtcaggatat gtggttctgg taggagacga gaacctaaaa
     6121 cagttcccgc ctccgtctga atttttgctt tcggtttgga accgaagccg cgcgtcttgt
     6181 ctgctgcagc gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg
     6241 tctgaaaatt agggccagac tgttaccact cccttaagtt tgaccttagg tcactggaaa
     6301 gatgtcgagc ggatcgctca caaccagtcg gtagatgtca agaagagacg ttgggttacc
     6361 ttctgctctg cagaatggcc aacctttaac gtcggatggc cgcgagacgg cacctttaac
     6421 cgagacctca tcacccaggt taagatcaag gtcttttcac ctggcccgca tggacaccca
     6481 gaccaggtcc cctacatcgt gacctgggaa gccttggctt ttgacccccc tccctgggtc
     6541 aagccctttg tacaccctaa gcctccgcct cctcttcctc catccgcccc gtctctcccc
     6601 cttgaacctc ctcgttcgac cccgcctcga tcctcccttt atccagccct cactccttct
     6661 ctaggcgccg gaattcgcgg ccgtgacaag agttactaac agcccctctc tccaagctca
     6721 cttacaggct ctctacttag tccagcacga agtctggaga cctctggcgg cagcctacca
     6781 agaacaactg gaccgaccgg tggtacctca cccttaccga gtcggcgaca cagtgtgggt
     6841 ccgccgacac cagactaaga acctagaacc tcgctggaaa ggaccttaca cagtcctgct
     6901 gaccaccccc accgccctca aagtagacgg catcgcagct tggatacacg ccgcccacgt
     6961 gaaggctgcc gaccccgggg gtggaccatc ctctagactg acgcggccgc tacaattcga
     7021 ttgtatgtta gggactgtga agatggaagg gcatgagagc aacgactgga acagctacta
     7081 cgcggacacg caggaggcct actcctctgt ccctgtcagc aacatgaact ccggcctggg
     7141 ctctatgaac tccatgaaca cctacatgac catgaacacc atgaccacga gcggcaacat
     7201 gaccccggct tccttcaaca tgtcctacgc caacacgggc ttaggggccg gcctgagtcc
     7261 cggtgctgtg gctggcatgc caggggcctc tgcaggcgcc atgaacagca tgactgcggc
     7321 gggcgtcacg gccatgggta cggcgctgag cccgggaggc atgggctcca tgggcgcgca
     7381 gcccgccacc tccatgaacg gcctgggtcc ctacgccgcc gccatgaacc cgtgcatgag
     7441 tcccatggcg tacgcgccgt ccaacctggg ccgcagccgc gcggggggcg gcggcgacgc
     7501 caagacattc aagcgcagct accctcacgc caagccgcct tactcctaca tctcgctcat
     7561 cacgatggcc atccagcagg cgcccagcaa gatgctcacg ctgagcgaga tctaccagtg
     7621 gatcatggac ctcttcccct attaccgcca gaaccagcag cgctggcaga actccatccg
     7681 cc
//
