LOCUS       sequence-106000-        7361 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 7361)
  AUTHORS   Park IH, Zhao R, West JA, Yabuuchi A, Huo H, Ince TA, Lerou PH, 
            Lensch MW, Daley GQ
  TITLE     Reprogramming of human somatic cells to pluripotency with defined 
            factors.
  JOURNAL   Nature. 2008 Jan 10. 451(7175):141-6.
  PUBMED    18157115
REFERENCE   2  (bases 1 to 7361)
  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: "Nature. 
            2008 Jan 10. 451(7175):141-6."
COMMENT     Sequence Label: pMIG-hSOX2
FEATURES             Location/Qualifiers
     source          1..7361
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(161..180)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      974..1562
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1463..1482
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1716..1733
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1850..1871
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1886..1891,1892..1909,1910..1916)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1924..1940
                     /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)."
     LTR             2102..2616
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     CDS             complement(join(2671..3384,3385..3387,3388..3390))
                     /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(2716..2737)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3065..3084
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3324..3345
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    3395..3856
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     3783..3800
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(4927..4950)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(4967..5383)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(4983..4999)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(5015..5037)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    5450..5791
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             5855..6371
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(6571..6590)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(6718..6740)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6934..6953)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7053..7075
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7113..7131)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7199..7303
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7304..7361,1..11,12..803)
                     /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"
ORIGIN
        1 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
       61 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
      121 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
      181 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
      241 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
      301 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
      361 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
      421 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
      481 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
      541 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
      601 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
      661 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
      721 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
      781 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
      841 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
      901 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
      961 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     1021 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     1081 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     1141 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     1201 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     1261 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     1321 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     1381 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     1441 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     1501 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     1561 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     1621 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     1681 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     1741 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     1801 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     1861 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     1921 gaattgtgag cggataacaa tttcacaatt ctccccagct ctttctgcgc tgtccataat
     1981 gaactatttc aggagtattg gagtatgtgg attcaaaaca gtggctaatt gagtcaggat
     2041 tacttaatat tcgactccta ccctcagaat gttgttctcc aacttcaaga aactgttacc
     2101 catgaaagac ccccgctgac gggtagtcaa tcactcagag gagaccctcc caaggaacag
     2161 cgagaccaca agtcggatgc aactgcaaga gggtttattg gatacacggg tacccgggcg
     2221 acgcagtcta tcggaggact ggcgcgccga gtgaggggtt gtgggctctt ttattgagct
     2281 cggggagcag aagcgcgcga acagaagcga gaagcgaact gattggttag ttcaaataag
     2341 gcacagggtc atttcaggtc cttggggcac cctggaaaca tctgatggtt ctctagaaac
     2401 tgctgagggc gggaccgcat ctggggacca tctgttcttg gccctgagcc ggggcaggaa
     2461 ctgcttacca cagatatcct gtttggccca tattctgctg tctctctgtt cctaaccttg
     2521 atctgaactt ctctattctc agttatgtat tttccatgcc ttgcaaaatg gcgttactta
     2581 agctagcttg ccaaacctac aggtggggtc tttcattccc ccctttttct ggagactaaa
     2641 taaaatcttt tattttatcg cgatgtcgac ttacttgtac agctcgtcca tgccgagagt
     2701 gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc
     2761 tttgctcagg gcggactggg tgctcaggta gtggttgtcg ggcagcagca cggggccgtc
     2821 gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt
     2881 gtggcggatc ttgaagttcg ccttgatgcc gttcttctgc ttgtcggcca tgatatagac
     2941 gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa
     3001 gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc
     3061 gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc
     3121 gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca
     3181 ctgcacgccg taggtcaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt
     3241 ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccaga
     3301 cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc
     3361 ggtgaacagc tcctcgccct tgctcaccat ggtattatca tcgtgttttt caaaggaaaa
     3421 ccacgtcccc gtggttcggg gggcctagac gttttttaac ctcgactaaa cacatgtaaa
     3481 gcatgtgcac cgaggcccca gatcagatcc catacaatgg ggtaccttct gggcatcctt
     3541 cagccccttg ttgaatacgc ttgaggtgag ccatttgact ctttccacaa ctatccaact
     3601 cacaacgtgg cactggggtt gtgccgcctt tgcaggtgta tcttatacac gtggcttttg
     3661 gccgcagagg cacctgtcgc caggtggggg gttccgctgc ctgcaaaggg tcgctacaga
     3721 cgttgtttgt cttcaagaag cttccagagg aactgcttcc ttcacgacat tcaacagacc
     3781 ttgcattcct ttggcgagag gggaaagacc cctaggaatg ctcgtcaaga agacagggcc
     3841 aggtttccgg gccctcacat tgccaaaaga cggcaatatg gtggaaaata acgtcgacga
     3901 tgtctcccag tgagtcggtt agtttgggtg gccgtagcct tgaggtacca gagatctctc
     3961 gagtcacatg tgtgagaggg gcagtgtgcc gttaatggcc gtgccgggca ccgggccgct
     4021 ctggtagtgc tgggacatgt gaagtctgct gggggcggcg ggttccggca cctcggcgcc
     4081 ggggagatac atgctgatca tgtcccggag gtccccggcc tggcagggcg ccctggagtg
     4141 ggaggaagag gtaaccacag gggggctgga gctggcctcg gacttgacca ccgaacccat
     4201 ggagccaaga gccatgccag gggtgccctg ctgcgagtag gacatgctgt aggtgggcga
     4261 gccgttcatg taggtctgcg agctggtcat ggagttgtac tgcagggcgc tcacgtcgta
     4321 gcggtgcatg ggctgcatct gcgctgcgcc gtgcgcattg aggcccgggt gctgcgggta
     4381 gcccagctgg tcctgcatca tgctgtagct gccgttgctc cagccgttca tgtgcgcgta
     4441 actgtccatg cgctggttca cgcccgcgcc caggccggcg cccaccccga ccccgctcgc
     4501 catgctattg ccgccggggg ccagcagccc gccgggcagc gtgtacttat ccttcttcat
     4561 gagcgtcttg gttttccgcc ggggccggta tttataatcc gggtgctcct tcatgtgcag
     4621 cgctcgcagc cgcttagcct cgtcgatgaa cggccgcttc tccgtctccg acaaaagttt
     4681 ccactcggcg cccaggcgct tgctgatctc cgagttgtgc atcttggggt tctcctgggc
     4741 catcttgcgc cgctgcccgc gggaccacac catgaaggca ttcatgggcc gcttgacgcg
     4801 gtccgggctg tttttctggt tgccgccggc cgccgccgcg gtggagttgc cgccgccgcc
     4861 ccccgaagtt tgctgcgggc ccggcggctt cagctccgtc tccatcatgt tgtacatgaa
     4921 ttcgatctta tcgtcgtcat ccttgtaatc catggtggac aattccggcg cctagagaag
     4981 gagtgagggc tggataaagg gaggatcgag gcggggtcga acgaggaggt tcaaggggga
     5041 gagacggggc ggatggagga agaggaggcg gaggcttagg gtgtacaaag ggcttgaccc
     5101 agggaggggg gtcaaaagcc aaggcttccc aggtcacgat gtaggggacc tggtctgggt
     5161 gtccatgcgg gccaggtgaa aagaccttga tcttaacctg ggtgatgagg tctcggttaa
     5221 aggtgccgtc tcgcggccat ccgacgttaa aggttggcca ttctgcagag cagaaggtaa
     5281 cccaacgtct cttcttgaca tctaccgact ggttgtgagc gatccgctcg acatctttcc
     5341 agtgacctaa ggtcaaactt aagggagtgg taacagtctg gccctaattt tcagacaaat
     5401 acagaaacac agtcagacag agacaacaca gaacgatgct gcagcgctgc agcagacaag
     5461 acgcgcggct tcggttccaa accgaaagca aaaattcaga cggaggcggg aactgtttta
     5521 ggttctcgtc tcctaccaga accacatatc ctgacggggt cggattccac atcgactccc
     5581 ttcctcaggt cgggccacaa aaacggcccc caaagtccct gggacgtctc ccagggttgc
     5641 ggccgggtgt tcagaactcg tcagttccac cacgggtccg ccagatacag agctagttag
     5701 ctaactagta cagacgcagg cgcaaacatt agatgccggc acaaacacgc acaaagacag
     5761 agacagacac gaaacgaccg ctggccagct tacctcccgg cgggggggtc ggtggtccct
     5821 gggcaggggt ctccaaatct cggtggaacc tccaaatgaa agacccccga ggtgggcagt
     5881 caatcaatct gaggagaccc tcccaaggat cagcgagtcc acgattcgga tgcaaacagc
     5941 aagaggcttt attgggaata cgggtacccg ggcgacgcag tctatcggag gactggcgcg
     6001 ccgagtgagg ggttgtgggc tcttttattg agctcgggga gcagaagcgc gcgaacagaa
     6061 gcgagaagcg aactgattgg ttagttcaaa taaggcacag ggtcatttca ggtccttggg
     6121 gcaccctgga aacatctgat ggttctctag aaactgctga gggcgggacc gcatctgggg
     6181 accatctgtt cttggccctg agccggggca ggaactgctt accacagata tcctgtttgg
     6241 cccatattct gctgtctctc tgttcctaac cttgatctga acttctctat tctcagttat
     6301 gtattttcca tgccttgcaa aatggcgtta cttaagctag cttgccaaac ctacaggtgg
     6361 ggtctttcat tccccccttt ttctggagac taaataaaat cttttatttt atctatggct
     6421 cgtactctat aggcttcagc tggtgatatt gttgagtcaa aactagagcc tggaccactg
     6481 atatcctgtc tttaacaaat tggactaatc gcctctacgc cggacgcatc gtggccggca
     6541 tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag
     6601 atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc
     6661 ccgtggccgg gggactgttg ggcgccatct ccttgcatgc accattcctt gcgccgtcgt
     6721 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     6781 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     6841 gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg
     6901 cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     6961 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     7021 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     7081 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     7141 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     7201 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     7261 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     7321 ccgtgtcgcc cttattccct tttttgcggc attttgcctt c
//
