LOCUS Exported 8740 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Dox-inducible polycistronic lentiviral vector expressing mouse Sox2, Klf4. ACCESSION . VERSION . KEYWORDS pHAGE2-TetOminiCMV-SK SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8740) AUTHORS Velychko S, Adachi K, Kim KP, Hou Y, MacCarthy CM, Wu G, Scholer HR TITLE Excluding Oct4 from Yamanaka Cocktail Unleashes the Developmental Potential of iPSCs. JOURNAL Cell Stem Cell. 2019 Oct 30. pii: S1934-5909(19)30423-0. doi: 10.1016/j.stem.2019.10.002. PUBMED 31708402 REFERENCE 2 (bases 1 to 8740) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8740 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 1..634 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 681..806 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1303..1536 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1721..1765 /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 2058..2175 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" protein_bind 2251..2521 /label=tetracycline response element /bound_moiety="tetracycline repressor TetR" /note="contains seven copies of the tetracycline operator tetO" primer_bind 2589..2613 /label=LNCX /note="Human CMV promoter, forward primer" CDS 3657..3710 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" misc_feature 5182..5770 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5235..5255) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5653..5664) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 5918..6098 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(6145..6163) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 6231..6335 /gene="bla" /label=AmpR promoter CDS 6336..7196 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(6554..6573) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7367..7955 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7856..7875 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 8109..8126 /label=L4440 /note="L4440 vector, forward primer" primer_bind 8218..8237 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" protein_bind 8366..8387 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 8402..8432 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8440..8456 /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)." primer_bind 8445..8467 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 8464..8480 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 8464..8480 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 8530..8549 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg 2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa 2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca 2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtcca 2221 caccctaact gacacactcg agtttaccac tccctatcag tgatagagaa aagtgaaagt 2281 cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta 2341 tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt 2401 gaaagtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac 2461 tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag 2521 aaaagtgaaa gtcgagctcg gtacccgggt cgaggtaggc atgtacggtg ggaggcctat 2581 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt 2641 gacctccata gaagacaccg ggaccgatcc agcctgcggc cgccatgtat aacatgatgg 2701 agacggagct gaagccgccg ggcccgcagc aagcttcggg gggcggcggc ggaggaggca 2761 acgccacggc ggcggcaacc ggcggcaacc agaagaacag cccggaccgc gtcaagaggc 2821 ccatgaacgc cttcatggta tggtcccggg ggcagcggcg taagatggcc caggagaacc 2881 ccaagatgca caactcggag atcagcaagc gcctgggcgc ggagtggaaa cttttgtccg 2941 agaccgagaa gcggccgttc atcgacgagg ccaagcggct gcgcgctctg cacatgaagg 3001 agcacccgga ttataaatac cggccgcggc ggaaaaccaa gacgctcatg aagaaggata 3061 agtacacgct tcccggaggc ttgctggccc ccggcgggaa cagcatggcg agcggggttg 3121 gggtgggcgc cggcctgggt gcgggcgtga accagcgcat ggacagctac gcgcacatga 3181 acggctggag caacggcagc tacagcatga tgcaggagca gctgggctac ccgcagcacc 3241 cgggcctcaa cgctcacggc gcggcacaga tgcaaccgat gcaccgctac gacgtcagcg 3301 ccctgcagta caactccatg accagctcgc agacctacat gaacggctcg cccacctaca 3361 gcatgtccta ctcgcagcag ggcacccccg gtatggcgct gggctccatg ggctctgtgg 3421 tcaagtccga ggccagctcc agcccccccg tggttacctc ttcctcccac tccagggcgc 3481 cctgccaggc cggggacctc cgggacatga tcagcatgta cctccccggc gccgaggtgc 3541 cggagcccgc tgcgcccagt agactgcaca tggcccagca ctaccagagc ggcccggtgc 3601 ccggcacggc cattaacggc acactgcccc tgtcgcacat ggcatgcggc tccggcgagg 3661 gcaggggaag tcttctaaca tgcggggacg tggaggaaaa tcccggccca ctcgagatga 3721 ggcagccacc tggcgagtct gacatggctg tcagcgacgc tctgctcccg tccttctcca 3781 cgttcgcgtc cggcccggcg ggaagggaga agacactgcg tccagcaggt gccccgacta 3841 accgttggcg tgaggaactc tctcacatga agcgacttcc cccacttccc ggccgcccct 3901 acgacctggc ggcgacggtg gccacagacc tggagagtgg cggagctggt gcagcttgca 3961 gcagtaacaa cccggccctc ctagcccgga gggagaccga ggagttcaac gacctcctgg 4021 acctagactt tatcctttcc aactcgctaa cccaccagga atcggtggcc gccaccgtga 4081 ccacctcggc gtcagcttca tcctcgtctt ccccagcgag cagcggccct gccagcgcgc 4141 cctccacctg cagcttcagc tatccgatcc gggccggggg tgacccgggc gtggctgcca 4201 gcaacacagg tggagggctc ctctacagcc gagaatctgc gccacctccc acggccccct 4261 tcaacctggc ggacatcaat gacgtgagcc cctcgggcgg cttcgtggct gagctcctgc 4321 ggccggagtt ggacccagta tacattccgc cacagcagcc tcagccgcca ggtggcgggc 4381 tgatgggcaa gtttgtgctg aaggcgtctc tgaccacccc tggcagcgag tacagcagcc 4441 cttcggtcat cagtgttagc aaaggaagcc cagacggcag ccaccccgtg gtagtggcgc 4501 cctacagcgg tggcccgccg cgcatgtgcc ccaagattaa gcaagaggcg gtcccgtcct 4561 gcacggtcag ccggtcccta gaggcccatt tgagcgctgg accccagctc agcaacggcc 4621 accggcccaa cacacacgac ttccccctgg ggcggcagct ccccaccagg actaccccta 4681 cactgagtcc cgaggaactg ctgaacagca gggactgtca ccctggcctg cctcttcccc 4741 caggattcca tccccatccg gggcccaact accctccttt cctgccagac cagatgcagt 4801 cacaagtccc ctctctccat tatcaagagc tcatgccacc gggttcctgc ctgccagagg 4861 agcccaagcc aaagagggga agaaggtcgt ggccccggaa aagaacagcc acccacactt 4921 gtgactatgc aggctgtggc aaaacctata ccaagagttc tcatctcaag gcacacctgc 4981 gaactcacac aggcgagaaa ccttaccact gtgactggga cggctgtggg tggaaattcg 5041 cccgctcgga tgaactgacc aggcactacc gcaaacacac agggcaccgg ccctttcagt 5101 gccagaagtg cgacagggcc ttttccaggt cggaccacct tgccttacac atgaagaggc 5161 acttttaaat cgatagatcc taatcaacct ctggattaca aaatttgtga aagattgact 5221 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg 5281 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg 5341 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg 5401 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg 5461 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc 5521 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca 5581 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc 5641 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct 5701 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc 5761 gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca 5821 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatca 5881 cctgcaggac aggcgcgccc tgctttttgc ttgtactggg tctctctggt tagaccagat 5941 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt 6001 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc 6061 cctcagaccc ttttagtcag tgtggaaaat ctctagcacc cgggcgatta aggaaagggc 6121 tagatcattc ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca 6181 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc 6241 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct 6301 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg 6361 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg 6421 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc 6481 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca 6541 cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac 6601 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa 6661 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg 6721 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt 6781 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg 6841 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc 6901 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga 6961 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta 7021 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc 7081 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg 7141 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt 7201 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa 7261 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt 7321 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt 7381 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt 7441 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga 7501 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag 7561 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata 7621 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg 7681 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga 7741 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca 7801 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa 7861 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt 7921 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac 7981 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt 8041 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga 8101 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc 8161 tccccgcgcg ttggccgatt cattaatgca gcaagctcat ggctgactaa ttttttttat 8221 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt 8281 ttttggaggc ctaggctttt gcaaaaagct ccccgtggca cgacaggttt cccgactgga 8341 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg 8401 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc 8461 acacaggaaa cagctatgac atgattacga atttcacaaa taaagcattt ttttcactgc 8521 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaactgga 8581 taactcaagc taaccaaaat catcccaaac ttcccacccc ataccctatt accactgcca 8641 attacctgtg gtttcattta ctctaaacct gtgattcctc tgaattattt tcattttaaa 8701 gaaattgtat ttgttaaata tgtactacaa acttagtagt //