LOCUS Exported 6263 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Episomal reprogramming . ACCESSION . VERSION . KEYWORDS pSimple-miR302/367 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6263) AUTHORS Howden SE, Maufort JP, Duffin BM, Elefanty AG, Stanley EG, Thomson JA TITLE Simultaneous Reprogramming and Gene Correction of Patient Fibroblasts. JOURNAL Stem Cell Reports. 2015 Dec 8;5(6):1109-18. doi: 10.1016/j.stemcr.2015.10.009. Epub 2015 Nov 12. PUBMED 26584543 REFERENCE 2 (bases 1 to 6263) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6263 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 213..1375 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 444..1366 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1323..1343 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" primer_bind complement(2553..2570) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2559..2783 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2948..4061 /label=mini-oriP /note="internally truncated variant of the Epstein-Barr virus oriP replication origin (Tanaka et al., 1999)" primer_bind complement(4106..4122) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4106..4122) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4119..4141) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4130..4146 /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(4154..4184) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4199..4220 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4337..4354) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4508..5096) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4588..4607) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5267..6127) /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 5890..5909 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6128..6232) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg gaggctccgg tgcccgtcag tgggcagagc 241 gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct 301 agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc 361 ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca 421 acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct 481 ttacgggtta tggcccttgc gtgccttgaa ttacttccac ctggctccag tacgtgattc 541 ttgatcccga gctggagcca ggggcgggcc ttgcgcttta ggagcccctt cgcctcgtgc 601 ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg 661 cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc 721 gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caggatctgc acactggtat 781 ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc 841 gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg 901 gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct 961 ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctccagg 1021 gggctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag 1081 gaaaggggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc 1141 gtccaggcac ctcgattagt tctggagctt ttggagtacg tcgtctttag gttgggggga 1201 ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc 1261 ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat 1321 tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgaacacg 1381 tggtcgcggc caattctgca gtcgacggta ccgcgggccc gggatccgga attcccatcg 1441 atggcctcga gaaagcggcc gcagctgcta gcgtccctgc cttttaccct tctggaggag 1501 aacacgaatc tttgggaact agttcaggaa ggtaaaaaaa tttttttctt ctaaagttat 1561 gccattttgt tttctttctc ctcagctcta aatactctga agtccaaaga agttgtatgt 1621 tgggtgggct cccttcaact ttaacatgga agtgctttct gtgactttaa aagtaagtgc 1681 ttccatgttt tagtaggagt gaatccaatt tacttctcca aaatagaaca cgctaacctc 1741 atttgaaggg atcccctttg ctttaacatg ggggtacctg ctgtgtgaaa caaaagtaag 1801 tgcttccatg tttcagtgga ggtgtctcca agccagcaca ccttttgtta caaaattttt 1861 ttgttattgt gttttaaggt tactaagctt gttacaggtt aaaggattct aactttttcc 1921 aagactgggc tccccaccac ttaaacgtgg atgtacttgc tttgaaacta aagaagtaag 1981 tgcttccatg ttttggtgat ggtaagtctt ccttttacat ttttattatt tttttagaaa 2041 ataactttat tgtattgacc gcagctcata tatttaagct ttattttgta tttttacatc 2101 tgttaagggg ccccctctac tttaacatgg aggcacttgc tgtgacatga caaaaataag 2161 tgcttccatg tttgagtgtg gtggttccta cctaatcagc aattgagtta acgcccacac 2221 tgtgtgcagt tcttggctac aggccattac tgttgctaat atgcaactct gttgaatata 2281 aattggaatt gcactttagc aatggtgatg gattgttaag ccaatgacag aatttaaacc 2341 acagacttac tttgatagca ctcttaatgg tataacttct tctcccattt tatgtctctc 2401 tttatgtttt ttcttatgtt tcctttttgt tttcaagaga gagctatctt ttagatctcc 2461 agtatccttt tcctcttcag tagagatttc tatatcttga aataaataaa aaagctctcc 2521 gttaagacat gggtctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc 2581 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt 2641 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct 2701 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc 2761 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctcgaccga 2821 tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg 2881 tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccctggccgg 2941 ggtcccgcag gaaaaggaca agcagcgaaa attcacgccc ccttgggagg tggcggcata 3001 tgcaaaggat agcactccca ctctactact gggtatcata tgctgactgt atatgcatga 3061 ggatagcata tgctacccgg atacagatta ggatagcata tactacccag atatagatta 3121 ggatagcata tgctacccag atatagatta ggatagccta tgctacccag atataaatta 3181 ggatagcata tactacccag atatagatta ggatagcata tgctacccag atatagatta 3241 ggatagccta tgctacccag atatagatta ggatagcata tgctacccag atatagatta 3301 ggatagcata tgctatccag atatttgggt agtatatgct acccagatat aaattaggat 3361 agcatatact accctaatct ctattaggat agcatatgct acccggatac agattaggat 3421 agcatatact acccagatat agattaggat agcatatgct acccagatat agattaggat 3481 agcctatgct acccagatat aaattaggat agcatatact acccagatat agattaggat 3541 agcatatgct acccagatat agattaggat agcctatgct acccagatat agattaggat 3601 agcatatgct atccagatat ttgggtagta tatgctatcc atggcaacat tagcccaccg 3661 tgctctcagc gacctcgtga atatgaggac caacaaccct gtgcttggcg ctcaggcgca 3721 agtgtgtgta atttgtcctc cagatcgcag caatcgcgcc cctatcttgg cccgcccacc 3781 tacttatgca ggtattcccc ggggtgccat tagtggtttt gtgggcaagt ggtttgaccg 3841 cagtggttag cggggttaca atcagccaag ttattacacc cttattttac agtccaaaac 3901 cgcagggcgg cgtgtggggg ctgacgcgat cgctgttcct taggaccctt ttactaaccc 3961 taattcgata gcatatgctt cccgttgggt aacatatgct attgaattag ggttagtctg 4021 gatagtatat actactaccc gggaagcata tgctacccgt ttagggttta ccgtcgacct 4081 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc 4141 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat 4201 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 4261 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 4321 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 4381 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 4441 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 4501 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 4561 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 4621 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 4681 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg 4741 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 4801 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 4861 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 4921 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc 4981 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 5041 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 5101 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 5161 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa 5221 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa 5281 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc 5341 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga 5401 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa 5461 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt 5521 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg 5581 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 5641 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg 5701 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 5761 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt 5821 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 5881 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac 5941 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac 6001 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag 6061 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa 6121 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga 6181 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc 6241 cccgaaaagt gccacctgac gtc //