LOCUS Exported 8350 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Dox-inducible polycistronic lentiviral vector expressing mouse Oct4, Sox2. ACCESSION . VERSION . KEYWORDS pHAGE2-TetOminiCMV-OS SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8350) 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 8350) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8350 /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" 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" misc_feature 4792..5380 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4845..4865) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5263..5274) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 5528..5708 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(5755..5773) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5841..5945 /gene="bla" /label=AmpR promoter CDS 5946..6806 /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(6164..6183) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6977..7565 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7466..7485 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7719..7736 /label=L4440 /note="L4440 vector, forward primer" primer_bind 7828..7847 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" protein_bind 7976..7997 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 8012..8042 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8050..8066 /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 8055..8077 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 8074..8090 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 8074..8090 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 8140..8159 /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 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 gaacttctct ctgttaaagc 3781 aagcaggaga 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 acatgtaaat cgatagatcc taatcaacct 4801 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 4861 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 4921 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 4981 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 5041 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 5101 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 5161 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt 5221 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 5281 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 5341 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat 5401 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg 5461 gctaattcac tcccaacgaa gacaagatca cctgcaggac aggcgcgccc tgctttttgc 5521 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg 5581 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg 5641 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat 5701 ctctagcacc cgggcgatta aggaaagggc tagatcattc ttgaagacga aagggcctcg 5761 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg 5821 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa 5881 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga 5941 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc 6001 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg 6061 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc 6121 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat 6181 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg 6241 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag 6301 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa 6361 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc 6421 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca 6481 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc 6541 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc 6601 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg 6661 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta 6721 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag 6781 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga 6841 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc 6901 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa 6961 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa 7021 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc 7081 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt 7141 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc 7201 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac 7261 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca 7321 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg 7381 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag 7441 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt 7501 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat 7561 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc 7621 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt 7681 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag 7741 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca 7801 gcaagctcat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc 7861 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct 7921 ccccgtggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg 7981 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt 8041 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac atgattacga 8101 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 8161 atgtatctta tcatgtctgg atcaactgga taactcaagc taaccaaaat catcccaaac 8221 ttcccacccc ataccctatt accactgcca attacctgtg gtttcattta ctctaaacct 8281 gtgattcctc tgaattattt tcattttaaa gaaattgtat ttgttaaata tgtactacaa 8341 acttagtagt //