LOCUS Exported 8263 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pLOVE empty vector SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8263) AUTHORS Blelloch R, Venere M, Yen J, Ramalho-Santos M TITLE Generation of Induced Pluripotent Stem Cells in the Absence of Drug Selection. JOURNAL Cell Stem Cell. 2007 Sep 13. ():. PUBMED 18371358 REFERENCE 2 (bases 1 to 8263) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8263 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(56..75) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 247..626 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 628..826 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" LTR 844..1024 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1071..1196 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1695..1928 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2113..2157 /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 2455..2572 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" enhancer 2638..2941 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2942..3145 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" protein_bind 3147..3165 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 3168..3186 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 3227..3351 /gene="mutant version of attR" /label=attR1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" promoter 3376..3406 /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" CDS 3460..4119 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(3526..3545) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" CDS 4461..4766 /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /label=ccdB /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" primer_bind 4645..4664 /label=ccdB-fwd /note="ccdB gene, forward primer" protein_bind complement(4807..4931) /gene="mutant version of attR" /label=attR2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 4984..5025 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" protein_bind 5063..5096 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 5152..5740 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5205..5225) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5623..5634) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(5743..5759) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5744..5760) /label=pBluescriptKS /note="For pBluescript vector" LTR 6269..6449 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" rep_origin complement(6511..7099) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6591..6610) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7270..8130) /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 7893..7912 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8131..8235) /gene="bla" /label=AmpR promoter ORIGIN 1 gcttaagcgg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa 61 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg 121 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca 181 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata 241 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc 301 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac 361 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa 421 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag 481 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc 541 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct 601 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg 661 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt 721 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga 781 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt 841 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac 901 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg 961 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct 1021 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac 1081 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta 1141 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta 1201 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga 1261 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta 1321 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat 1381 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt 1441 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag 1501 agcaaaacaa aagtaagacc accgcacagc aagcggccgg ccgcgctgat cttcagacct 1561 ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa 1621 attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa 1681 agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg 1741 ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag 1801 cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc 1861 tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa 1921 cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg 1981 aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag 2041 tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa 2101 aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg 2161 aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga 2221 ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag 2281 ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc 2341 gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac 2401 ggatcggcac tgcgtgcgcc aattctgcag acaaatggca gtattcatcc acaattttaa 2461 aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac 2521 agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta 2581 ttacagggac agcagagatc cagtttggtt agtaccgggc ccgctctagc agttccgcgt 2641 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac 2701 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg 2761 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag 2821 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat 2881 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat 2941 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt 3001 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga 3061 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg 3121 gtgggaggtc tatataagca gagctctccc tatcagtgat agagatctcc ctatcagtga 3181 tagagatcgt cgactagtcc agtgtggtgg aattctgcag atatcaacaa gtttgtacaa 3241 aaaagctgaa cgagaaacgt aaaatgatat aaatatcaat atattaaatt agattttgca 3301 taaaaaacag actacataat actgtaaaac acaacatatc cagtcactat ggcggccgca 3361 ttaggcaccc caggctttac actttatgct tccggctcgt ataatgtgtg gattttgagt 3421 taggatccgt cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga 3481 tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca 3541 gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc 3601 gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg 3661 aatgctcatc cggaattccg tatggcaatg aaagacggtg agctggtgat atgggatagt 3721 gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt 3781 gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac 3841 ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc 3901 aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc 3961 gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg 4021 gcgattcagg ttcatcatgc cgtctgtgat ggcttccatg tcggcagaat gcttaatgaa 4081 ttacaacagt actgcgatga gtggcagggc ggggcgtaaa gatctggatc cggcttacta 4141 aaagccagat aacagtatgc gtatttgcgc gctgattttt gcggtataag aatatatact 4201 gatatgtata cccgaagtat gtcaaaaaga ggtgtgctat gaagcagcgt attacagtga 4261 cagttgacag cgacagctat cagttgctca aggcatatat gatgtcaata tctccggtct 4321 ggtaagcaca accatgcaga atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga 4381 aaatcaggaa gggatggctg aggtcgcccg gtttattgaa atgaacggct cttttgctga 4441 cgagaacagg gactggtgaa atgcagttta aggtttacac ctataaaaga gagagccgtt 4501 atcgtctgtt tgtggatgta cagagtgata ttattgacac gcccgggcga cggatggtga 4561 tccccctggc cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg 4621 tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggtct 4681 ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca 4741 ttaacctgat gttctgggga atataaatgt caggctccct tatacacagc cagtctgcag 4801 gtcgaccata gtgactggat atgttgtgtt ttacagtatt atgtagtctg ttttttatgc 4861 aaaatctaat ttaatatatt gatatttata tcattttacg tttctcgttc agctttcttg 4921 tacaaagtgg ttgatatcca gcacagtggc ggccgctcga gtctagaggg cccgcggttc 4981 gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac cggttagtaa 5041 tgacaattcg tcgagggacc taataacttc gtatagcata cattatacga agttatacat 5101 gtttaagggt tccggttcca ctaggtacaa ttcgatatca agcttatcga taatcaacct 5161 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 5221 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 5281 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 5341 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 5401 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 5461 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 5521 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt 5581 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 5641 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 5701 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgacctcga 5761 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga 5821 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt 5881 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa 5941 ggggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat 6001 ctaccacaca caaggctact tccctgattg gcagaactac acaccagggc cagggatcag 6061 atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag agaaggtaga 6121 agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc atgggatgga 6181 tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat ttcatcacat 6241 ggcccgagag ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg 6301 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt 6361 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc 6421 cttttagtca gtgtggaaaa tctctagcag catgtgagca aaaggccagc aaaaggccag 6481 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 6541 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 6601 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 6661 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag 6721 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 6781 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 6841 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 6901 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt 6961 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 7021 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg 7081 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 7141 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta 7201 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg 7261 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg 7321 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc 7381 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc 7441 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc 7501 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag 7561 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat 7621 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg 7681 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt 7741 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag 7801 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg 7861 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt 7921 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct 7981 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac 8041 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat 8101 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat 8161 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca 8221 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gac //