LOCUS Exported 7361 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pMIG-hSOX2 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7361 /organism="synthetic DNA construct" /mol_type="other DNA" 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 1886..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(2671..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 7304..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 //