LOCUS Exported 9874 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Dox-inducible polycistronic reprogramming lentiviral vector expressing mouse Oct4, Sox2, Klf4. ACCESSION . VERSION . KEYWORDS pHAGE2-TetOminiCMV-OSK SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9874) 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 9874) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9874 /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" promoter 2554..2592 /label=minimal CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2589..2613 /label=LNCX /note="Human CMV promoter, forward primer" regulatory 2679..2688 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3756..3812 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS 4791..4844 /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 6316..6904 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(6369..6389) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6787..6798) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 7052..7232 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(7279..7297) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 7365..7469 /gene="bla" /label=AmpR promoter CDS 7470..8330 /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(7688..7707) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 8501..9089 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8990..9009 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 9243..9260 /label=L4440 /note="L4440 vector, forward primer" primer_bind 9352..9371 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" protein_bind 9500..9521 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 9536..9566 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 9574..9590 /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 9579..9601 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 9598..9614 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 9598..9614 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 9664..9683 /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 gtgtacggtg ggaggcctat 2581 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt 2641 gacctccata gaagacaccg ggaccgatcc agcctgcggc cgccatggct ggacacctgg 2701 cttcagactt cgccttctca cccccaccag gtgggggtga tgggtcagca gggctggagc 2761 cgggctgggt ggatcctcga acctggctaa gcttccaagg gcctccaggt gggcctggaa 2821 tcggaccagg ctcagaggta ttggggatct ccccatgtcc gcccgcatac gagttctgcg 2881 gagggatggc atactgtgga cctcaggttg gactgggcct agtcccccaa gttggcgtgg 2941 agactttgca gcctgagggc caggcaggag cacgagtgga aagcaactca gagggaacct 3001 cctctgagcc ctgtgccgac cgccccaatg ccgtgaagtt ggagaaggtg gaaccaactc 3061 ccgaggagtc ccaggacatg aaagccctgc agaaggagct agaacagttt gccaagctgc 3121 tgaagcagaa gaggatcacc ttggggtaca cccaggccga cgtggggctc accctgggcg 3181 ttctctttgg aaaggtgttc agccagacca ccatctgtcg cttcgaggcc ttgcagctca 3241 gccttaagaa catgtgtaag ctgcggcccc tgctggagaa gtgggtggag gaagccgaca 3301 acaatgagaa ccttcaggag atatgcaaat cggagaccct ggtgcaggcc cggaagagaa 3361 agcgaactag cattgagaac cgtgtgaggt ggagtctgga gaccatgttt ctgaagtgcc 3421 cgaagccctc cctacagcag atcactcaca tcgccaatca gcttgggcta gagaaggatg 3481 tggttcgagt atggttctgt aaccggcgcc agaagggcaa aagatcaagt attgagtatt 3541 cccaacgaga agagtatgag gctacaggga cacctttccc agggggggct gtatcctttc 3601 ctctgccccc aggtccccac tttggcaccc caggctatgg aagcccccac ttcaccacac 3661 tctactcagt cccttttcct gagggcgagg cctttccctc tgttcccgtc actgctctgg 3721 gctctcccat gcattcaaac gctagcggca gcggcgccac aaacttctct ctgctaaagc 3781 aagcaggtga tgttgaagaa aaccccgggc ctgcatgcat gtataacatg atggagacgg 3841 agctgaagcc gccgggcccg cagcaagctt cggggggcgg cggcggagga ggcaacgcca 3901 cggcggcggc gaccggcggc aaccagaaga acagcccgga ccgcgtcaag aggcccatga 3961 acgccttcat ggtatggtcc cgggggcagc ggcgtaagat ggcccaggag aaccccaaga 4021 tgcacaactc ggagatcagc aagcgcctgg gcgcggagtg gaaacttttg tccgagaccg 4081 agaagcggcc gttcatcgac gaggccaagc ggctgcgcgc tctgcacatg aaggagcacc 4141 cggattataa ataccggccg cggcggaaaa ccaagacgct catgaagaag gataagtaca 4201 cgcttcccgg aggcttgctg gcccccggcg ggaacagcat ggcgagcggg gttggggtgg 4261 gcgccggcct gggtgcgggc gtgaaccagc gcatggacag ctacgcgcac atgaacggct 4321 ggagcaacgg cagctacagc atgatgcagg agcagctggg ctacccgcag cacccgggcc 4381 tcaacgctca cggcgcggca cagatgcaac cgatgcaccg ctacgacgtc agcgccctgc 4441 agtacaactc catgaccagc tcgcagacct acatgaacgg ctcgcccacc tacagcatgt 4501 cctactcgca gcagggcacc cccggtatgg cgctgggctc catgggctct gtggtcaagt 4561 ccgaggccag ctccagcccc cccgtggtta cctcttcctc ccactccagg gcgccctgcc 4621 aggccgggga cctccgggac atgatcagca tgtacctccc cggcgccgag gtgccggagc 4681 ccgctgcgcc cagtagactg cacatggccc agcactacca gagcggcccg gtgcccggca 4741 cggccattaa cggcacactg cccctgtcgc acatggcatg cggctccggc gagggcaggg 4801 gaagtcttct aacatgcggg gacgtggagg aaaatcccgg cccactcgag atgaggcagc 4861 cacctggcga gtctgacatg gctgtcagcg acgctctgct cccgtccttc tccacgttcg 4921 cgtccggccc ggcgggaagg gagaagacac tgcgtccagc aggtgccccg actaaccgtt 4981 ggcgtgagga actctctcac atgaagcgac ttcccccact tcccggccgc ccctacgacc 5041 tggcggcgac ggtggccaca gacctggaga gtggcggagc tggtgcagct tgcagcagta 5101 acaacccggc cctcctagcc cggagggaga ccgaggagtt caacgacctc ctggacctag 5161 actttatcct ttccaactcg ctaacccacc aggaatcggt ggccgccacc gtgaccacct 5221 cggcgtcagc ttcatcctcg tcttccccag cgagcagcgg ccctgccagc gcgccctcca 5281 cctgcagctt cagctatccg atccgggccg ggggtgaccc gggcgtggct gccagcaaca 5341 caggtggagg gctcctctac agccgagaat ctgcgccacc tcccacggcc cccttcaacc 5401 tggcggacat caatgacgtg agcccctcgg gcggcttcgt ggctgagctc ctgcggccgg 5461 agttggaccc agtatacatt ccgccacagc agcctcagcc gccaggtggc gggctgatgg 5521 gcaagtttgt gctgaaggcg tctctgacca cccctggcag cgagtacagc agcccttcgg 5581 tcatcagtgt tagcaaagga agcccagacg gcagccaccc cgtggtagtg gcgccctaca 5641 gcggtggccc gccgcgcatg tgccccaaga ttaagcaaga ggcggtcccg tcctgcacgg 5701 tcagccggtc cctagaggcc catttgagcg ctggacccca gctcagcaac ggccaccggc 5761 ccaacacaca cgacttcccc ctggggcggc agctccccac caggactacc cctacactga 5821 gtcccgagga actgctgaac agcagggact gtcaccctgg cctgcctctt cccccaggat 5881 tccatcccca tccggggccc aactaccctc ctttcctgcc agaccagatg cagtcacaag 5941 tcccctctct ccattatcaa gagctcatgc caccgggttc ctgcctgcca gaggagccca 6001 agccaaagag gggaagaagg tcgtggcccc ggaaaagaac agccacccac acttgtgact 6061 atgcaggctg tggcaaaacc tataccaaga gttctcatct caaggcacac ctgcgaactc 6121 acacaggcga gaaaccttac cactgtgact gggacggctg tgggtggaaa ttcgcccgct 6181 ccgatgaact gaccaggcac taccgcaaac acacagggca ccggcccttt cagtgccaga 6241 agtgcgacag ggccttttcc aggtcggacc accttgcctt acacatgaag aggcactttt 6301 aaatcgatag atcctaatca acctctggat tacaaaattt gtgaaagatt gactggtatt 6361 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat 6421 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct 6481 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct 6541 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc 6601 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg 6661 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc 6721 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac 6781 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg 6841 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc 6901 ccgcctggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt 6961 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag atcacctgca 7021 ggacaggcgc gccctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc 7081 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg 7141 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag 7201 acccttttag tcagtgtgga aaatctctag cacccgggcg attaaggaaa gggctagatc 7261 attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa 7321 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt 7381 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa 7441 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta 7501 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag 7561 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca 7621 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta 7681 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc 7741 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc 7801 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca 7861 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc 7921 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca 7981 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac 8041 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg 8101 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg 8161 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg 8221 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac 8281 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc 8341 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct 8401 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc 8461 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc 8521 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg 8581 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa 8641 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc 8701 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt 8761 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa 8821 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc 8881 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc 8941 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct 9001 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat 9061 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc 9121 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg 9181 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc 9241 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg 9301 cgcgttggcc gattcattaa tgcagcaagc tcatggctga ctaatttttt ttatttatgc 9361 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg 9421 aggcctaggc ttttgcaaaa agctccccgt ggcacgacag gtttcccgac tggaaagcgg 9481 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac 9541 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag 9601 gaaacagcta tgacatgatt acgaatttca caaataaagc atttttttca ctgcattcta 9661 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcaac tggataactc 9721 aagctaacca aaatcatccc aaacttccca ccccataccc tattaccact gccaattacc 9781 tgtggtttca tttactctaa acctgtgatt cctctgaatt attttcattt taaagaaatt 9841 gtatttgtta aatatgtact acaaacttag tagt //