LOCUS Exported 9736 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Dox-inducible polycistronic reprogramming lentiviral vector expressing mouse Oct4, Sox2, cMyc. ACCESSION . VERSION . KEYWORDS pHAGE2-TetOminiCMV-OSM SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9736) 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 9736) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9736 /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" CDS 4785..4844 /codon_start=1 /product="2A peptide from equine rhinitis A virus polyprotein" /label=E2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="QCTNYALLKLAGDVESNPGP" misc_feature 6178..6766 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(6231..6251) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6649..6660) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 6914..7094 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(7141..7159) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 7227..7331 /gene="bla" /label=AmpR promoter CDS 7332..8192 /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(7550..7569) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 8363..8951 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8852..8871 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 9105..9122 /label=L4440 /note="L4440 vector, forward primer" primer_bind 9214..9233 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" protein_bind 9362..9383 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 9398..9428 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 9436..9452 /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 9441..9463 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 9460..9476 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 9460..9476 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 9526..9545 /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 ctgcttgcat 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 acatgggtag tgggcaatgt actaactacg 4801 ctttgttgaa actcgctggc gatgttgaaa gtaaccccgg tcctatgccc ctcaacgtga 4861 acttcaccaa caggaactat gacctcgact acgactccgt acagccctat ttcatctgcg 4921 acgaggaaga gaatttctat caccagcaac agcagagcga gctgcagccg cccgcgccca 4981 gtgaggatat ctggaagaaa ttcgagctgc ttcccacccc gcccctgtcc ccgagccgcc 5041 gctccgggct ctgctctcca tcctatgttg cggtcgctac gtccttctcc ccaagggaag 5101 acgatgacgg cggcggtggc aacttctcca ccgccgatca gctggagatg atgaccgagt 5161 tacttggagg agacatggtg aaccagagct tcatctgcga tcctgacgac gagaccttca 5221 tcaagaacat catcatccag gactgtatgt ggagcggttt ctcagccgct gccaagctgg 5281 tctcggagaa gctggcctcc taccaggctg cgcgcaaaga cagcaccagc ctgagccccg 5341 cccgcgggca cagcgtctgc tccacctcca gcctgtacct gcaggacctc accgccgccg 5401 cgtccgagtg cattgacccc tcagtggtct ttccctaccc gctcaacgac agcagctcgc 5461 ccaaatcctg tacctcgtcc gattccacgg ccttctctcc ttcctcggac tcgctgctgt 5521 cctccgagtc ctccccacgg gccagccctg agcccctagt gctgcatgag gagacaccgc 5581 ccaccaccag cagcgactct gaagaagagc aagaagatga ggaagaaatt gatgtggtgt 5641 ctgtggagaa gaggcaaacc cctgccaaga ggtcggagtc gggctcatct ccatcccgag 5701 gccacagcaa acctccgcac agcccactgg tcctcaagag gtgccacgtc tccactcacc 5761 agcacaacta cgccgcaccc ccctccacaa ggaaggacta tccagctgcc aagagggcca 5821 agttggacag tggcagggtc ctgaagcaga tcagcaacaa ccgcaagtgc tccagcccca 5881 ggtcctcaga cacggaggaa aacgacaaga ggcggacaca caacgtcttg gaacgtcaga 5941 ggaggaacga gctgaagcgc agcttttttg ccctgcgtga ccagatccct gaattggaaa 6001 acaacgaaaa ggcccccaag gtagtgatcc tcaaaaaagc caccgcctac atcctgtcca 6061 ttcaagcaga cgagcacaag ctcacctctg aaaaggactt attgaggaaa cgacgagaac 6121 agttgaaaca caaactcgaa cagcttcgaa actctggtgc ataaatcgat agatcctaat 6181 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct 6241 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg 6301 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg 6361 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt 6421 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt 6481 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg 6541 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc 6601 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat 6661 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc 6721 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg tacctttaag 6781 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact 6841 ggaagggcta attcactccc aacgaagaca agatcacctg caggacaggc gcgccctgct 6901 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta 6961 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg 7021 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg 7081 gaaaatctct agcacccggg cgattaagga aagggctaga tcattcttga agacgaaagg 7141 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt 7201 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac 7261 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa 7321 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat 7381 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc 7441 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga 7501 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg 7561 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc 7621 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag 7681 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc 7741 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg 7801 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg 7861 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac 7921 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac 7981 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg 8041 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg 8101 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg 8161 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac 8221 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg 8281 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 8341 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 8401 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 8461 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt 8521 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 8581 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 8641 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 8701 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 8761 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 8821 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 8881 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 8941 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 9001 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct 9061 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg 9121 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt 9181 aatgcagcaa gctcatggct gactaatttt ttttatttat gcagaggccg aggccgcctc 9241 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa 9301 aaagctcccc gtggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa 9361 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc 9421 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgacatga 9481 ttacgaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac 9541 tcatcaatgt atcttatcat gtctggatca actggataac tcaagctaac caaaatcatc 9601 ccaaacttcc caccccatac cctattacca ctgccaatta cctgtggttt catttactct 9661 aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt taaatatgta 9721 ctacaaactt agtagt //