LOCUS Exported 7542 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS FUW-tetO-lox-hOCT4 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7542) AUTHORS Frank Soldner, Dirk Hockemeyer, Caroline Beard, Qing Gao, George W. Bell, Elizabeth G. Cook, Gunnar Hargus, Alexandra Blak, Oliver Cooper, Maisam Mitalipova, Ole Isacson, Rudolf Jaenisch TITLE Parkinson's Disease Patient-Derived Induced Pluripotent Stem Cells Free of Viral Reprogramming Factors JOURNAL Cell, Volume 136, Issue 5, 964-977, 6 March 2009 REFERENCE 2 (bases 1 to 7542) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7542 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 6..385 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 387..585 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 540..560 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 603..783 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 830..955 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1448..1681 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1866..1910 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 2208..2325 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" protein_bind 2384..2654 /label=tetracycline response element /bound_moiety="tetracycline repressor TetR" /note="contains seven copies of the tetracycline operator tetO" promoter 2687..2724 /label=minimal CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2721..2745 /label=LNCX /note="Human CMV promoter, forward primer" misc_feature 3931..4519 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3984..4004) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4402..4413) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(4522..4538) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4523..4539) /label=pBluescriptKS /note="For pBluescript vector" protein_bind complement(5043..5076) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." LTR 5090..5270 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(5296..5313) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 5302..5413 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin complement(5559..6147) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5639..5658) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6318..7178) /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 6941..6960 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7179..7283) /gene="bla" /label=AmpR promoter primer_bind complement(7358..7377) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca 61 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc 121 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat 181 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt 241 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc 301 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta 361 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg 421 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt 481 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac 541 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagcgcgt tttgcctgta 601 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc 661 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt 721 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta 781 gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg 841 caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac 901 gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat 961 taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa 1021 aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa 1081 tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc 1141 ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg 1201 tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga 1261 gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag 1321 gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac 1381 cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag 1441 tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag 1501 cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga 1561 acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca 1621 tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc 1681 tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta 1741 gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca 1801 gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc 1861 aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt 1921 ttaacataac aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg 1981 taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt 2041 caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa 2101 tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg aacggatcgg 2161 cactgcgtgc gccaattctg cagacaaatg gcagtattca tccacaattt taaaagaaaa 2221 ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata 2281 caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg 2341 gacagcagag atccagtttg gttaatgatc tcgagtttac cactccctat cagtgataga 2401 gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt 2461 ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt 2521 gatagagaaa agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag 2581 tcgagtttac cactccctat cagtgataga gaaaagtgaa agtcgagttt accactccct 2641 atcagtgata gagaaaagtg aaagtcgagc tcggtacccg ggtcgagtag gcgtgtacgg 2701 tgggaggcct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat 2761 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg cggccccgaa 2821 ttcatggcgg gacacctggc ttcggatttc gccttctcgc cccctccagg tggtggaggt 2881 gatgggccag gggggccgga gccgggctgg gttgatcctc ggacctggct aagcttccaa 2941 ggccctcctg gagggccagg aatcgggccg ggggttgggc caggctctga ggtgtggggg 3001 attcccccat gccccccgcc gtatgagttc tgtgggggga tggcgtactg tgggccccag 3061 gttggagtgg ggctagtgcc ccaaggcggc ttggagacct ctcagcctga gggcgaagca 3121 ggagtcgggg tggagagcaa ctccgatggg gcctccccgg agccctgcac cgtcacccct 3181 ggtgccgtga agctggagaa ggagaagctg gagcaaaacc cggaggagtc ccaggacatc 3241 aaagctctgc agaaagaact cgagcaattt gccaagctcc tgaagcagaa gaggatcacc 3301 ctgggatata cacaggccga tgtggggctc accctggggg ttctatttgg gaaggtattc 3361 agccaaacga ccatctgccg ctttgaggct ctgcagctta gcttcaagaa catgtgtaag 3421 ctgcggccct tgctgcagaa gtgggtggag gaagctgaca acaatgaaaa tcttcaggag 3481 atatgcaaag cagaaaccct cgtgcaggcc cgaaagagaa agcgaaccag tatcgagaac 3541 cgagtgagag gcaacctgga gaatttgttc ctgcagtgcc cgaaacccac actgcagcag 3601 atcagccaca tcgcccagca gcttgggctc gagaaggatg tggtccgagt gtggttctgt 3661 aaccggcgcc agaagggcaa gcgatcaagc agcgactatg cacaacgaga ggattttgag 3721 gctgctgggt ctcctttctc agggggacca gtgtcctttc ctctggcccc agggccccat 3781 tttggtaccc caggctatgg gagccctcac ttcactgcac tgtactcctc ggtccctttc 3841 cctgaggggg aagcctttcc ccctgtctcc gtcaccactc tgggctctcc catgcattca 3901 aactgagaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa 3961 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta 4021 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa 4081 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg 4141 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc 4201 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc 4261 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg 4321 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg 4381 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg 4441 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc 4501 ctttgggccg cctccccgca tcgataccgt cgacctcgag acctagaaaa acatggagca 4561 atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag 4621 gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag 4681 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac 4741 tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct 4801 gattggcaga actacacacc agggccaggg atcagatatc cactgacctt tggatggtgc 4861 tacaagctag taccagttga gcaagagaag gtagaagaag ccaatgaagg agagaacacc 4921 cgcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga agtattagag 4981 tggaggtttg acagccgcct agcatttcat cacatggccc gagagctgca tccggactcg 5041 agataacttc gtataatgta tgctatacga agttattccg gactgtactg ggtctctctg 5101 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc 5161 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg 5221 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gggcccgttt 5281 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct 5341 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg 5401 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc 5461 aggacagcaa gggggaggat tgggaagaca atagcaggca tgtgagcaaa aggccagcaa 5521 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 5581 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 5641 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 5701 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca 5761 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 5821 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 5881 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 5941 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga 6001 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 6061 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 6121 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 6181 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc 6241 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag 6301 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt 6361 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag 6421 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca 6481 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact 6541 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca 6601 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg 6661 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc 6721 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg 6781 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca 6841 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt 6901 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc 6961 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc 7021 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca 7081 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa 7141 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat 7201 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa 7261 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga 7321 tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat 7381 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca 7441 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg 7501 ttaggcgttt tgcgctgctt cgcgatgtac gggccagata ta //