LOCUS       Exported                7484 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pMIG-hOCT4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7484)
  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 7484)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7484
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     13..29
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             62..85
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    1279..1740
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1335..1352)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             1745..2464
                     /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(1790..1811)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2051..2070)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2398..2419
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2519..3033
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3195..3211
                     /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(3219..3249)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3264..3285
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3402..3419)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3573..4161)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3653..3672)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4332..5192)
                     /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     4955..4974
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5193..5297)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5365..5383
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5421..5443)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5543..5562
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5756..5778
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5906..5925
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6125..6641
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6705..7046
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             7113..45
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     7459..7481
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
ORIGIN
        1 cctcgatcct ccctttatcc agccctcact ccttctctag gcgccggaat tgtccaccat
       61 ggattacaag gatgacgacg ataagatcga attcatggcg ggacacctgg cttcggattt
      121 cgccttctcg ccccctccag gtggtggagg tgatgggcca ggggggccgg agccgggctg
      181 ggttgatcct cggacctggc taagcttcca aggccctcct ggagggccag gaatcgggcc
      241 gggggttggg ccaggctctg aggtgtgggg gattccccca tgccccccgc cgtatgagtt
      301 ctgtgggggg atggcgtact gtgggcccca ggttggagtg gggctagtgc cccaaggcgg
      361 cttggagacc tctcagcctg agggcgaagc aggagtcggg gtggagagca actccgatgg
      421 ggcctccccg gagccctgca ccgtcacccc tggtgccgtg aagctggaga aggagaagct
      481 ggagcaaaac ccggaggagt cccaggacat caaagctctg cagaaagaac tcgagcaatt
      541 tgccaagctc ctgaagcaga agaggatcac cctgggatat acacaggccg atgtggggct
      601 caccctgggg gttctatttg ggaaggtatt cagccaaacg accatctgcc gctttgaggc
      661 tctgcagctt agcttcaaga acatgtgtaa gctgcggccc ttgctgcaga agtgggtgga
      721 ggaagctgac aacaatgaaa atcttcagga gatatgcaaa gcagaaaccc tcgtgcaggc
      781 ccgaaagaga aagcgaacca gtatcgagaa ccgagtgaga ggcaacctgg agaatttgtt
      841 cctgcagtgc ccgaaaccga cactgcagca gatcagccac atcgcccagc agcttgggct
      901 cgagaaggat gtggtccgag tgtggttctg taaccggcgc cagaagggca agcgatcaag
      961 cagcgactat gcacaacgag aggattttga ggctgctggg tctcctttct cagggggacc
     1021 agtgtccttt cctctggccc cagggcccca ttttggtacc ccaggctatg ggagccctca
     1081 cttcactgca ctgtactcct cggtcccttt ccctgagggg gaagcctttc cccctgtctc
     1141 cgtcaccact ctgggctctc ccatgcattc aaactgaaga tctctggtac ctcaaggcta
     1201 cggccaccca aactaaccga ctcactggga gacatcgtcg acgttatttt ccaccatatt
     1261 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc
     1321 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc
     1381 agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg
     1441 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc
     1501 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa
     1561 atggctcacc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg
     1621 tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa
     1681 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa
     1741 taccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     1801 ggacggcgac gtaaacggcc acaagttcag cgtgtctggc gagggcgagg gcgatgccac
     1861 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     1921 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     1981 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     2041 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     2101 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     2161 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     2221 gaacggcatc aaggcgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     2281 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     2341 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     2401 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     2461 gtaagtcgac atcgcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat
     2521 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     2581 gaaaatacat aactgagaat agagaagttc agatcaaggt taggaacaga gagacagcag
     2641 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     2701 cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat cagatgtttc
     2761 cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg
     2821 cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc
     2881 tcactcggcg cgccagtcct ccgatagact gcgtcgcccg ggtacccgtg tatccaataa
     2941 accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga
     3001 gtgattgact acccgtcagc gggggtcttt catgggtaac agtttcttga agttggagaa
     3061 caacattctg agggtaggag tcgaatatta agtaatcctg actcaattag ccactgtttt
     3121 gaatccacat actccaatac tcctgaaata gttcattatg gacagcgcag aaagagctgg
     3181 ggagaattgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     3241 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     3301 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     3361 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     3421 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3481 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3541 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3601 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3661 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3721 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3781 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3841 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3901 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3961 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4021 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4081 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4141 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4201 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4261 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4321 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4381 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4441 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4501 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4561 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4621 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4681 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4741 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4801 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4861 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4921 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4981 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5041 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5101 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5161 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5221 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5281 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     5341 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     5401 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     5461 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     5521 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     5581 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     5641 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     5701 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     5761 tcacgacgtt gtaaaacgac ggcgcaagga atggtgcatg caaggagatg gcgcccaaca
     5821 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
     5881 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
     5941 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggcgatta gtccaatttg
     6001 ttaaagacag gatatcagtg gtccaggctc tagttttgac tcaacaatat caccagctga
     6061 agcctataga gtacgagcca tagataaaat aaaagatttt atttagtctc cagaaaaagg
     6121 ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca
     6181 aggcatggaa aatacataac tgagaataga gaagttcaga tcaaggttag gaacagagag
     6241 acagcagaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
     6301 ccaagaacag atggtcccca gatgcggtcc cgccctcagc agtttctaga gaaccatcag
     6361 atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat
     6421 cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca
     6481 caacccctca ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt acccgtattc
     6541 ccaataaagc ctcttgctgt ttgcatccga atcgtggact cgctgatcct tgggagggtc
     6601 tcctcagatt gattgactgc ccacctcggg ggtctttcat ttggaggttc caccgagatt
     6661 tggagacccc tgcccaggga ccaccgaccc ccccgccggg aggtaagctg gccagcggtc
     6721 gtttcgtgtc tgtctctgtc tttgtgcgtg tttgtgccgg catctaatgt ttgcgcctgc
     6781 gtctgtacta gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt
     6841 tctgaacacc cggccgcaac cctgggagac gtcccaggga ctttgggggc cgtttttgtg
     6901 gcccgacctg aggaagggag tcgatgtgga atccgacccc gtcaggatat gtggttctgg
     6961 taggagacga gaacctaaaa cagttcccgc ctccgtctga atttttgctt tcggtttgga
     7021 accgaagccg cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt tgtctctgtc
     7081 tgactgtgtt tctgtatttg tctgaaaatt agggccagac tgttaccact cccttaagtt
     7141 tgaccttagg tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca
     7201 agaagagacg ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc
     7261 cgcgagacgg cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac
     7321 ctggcccgca tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt
     7381 ttgacccccc tccctgggtc aagccctttg tacaccctaa gcctccgcct cctcttcctc
     7441 catccgcccc gtctctcccc cttgaacctc ctcgttcgac cccg
//
