LOCUS Exported 7315 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Dox-inducible lentiviral vector expressing mouse Oct4. ACCESSION . VERSION . KEYWORDS pHAGE2-TetOminiCMV-Oct4 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7315) 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 7315) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7315 /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)" misc_feature 3757..4345 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3810..3830) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4228..4239) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 4493..4673 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(4720..4738) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4806..4910 /gene="bla" /label=AmpR promoter CDS 4911..5771 /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(5129..5148) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5942..6530 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6431..6450 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6684..6701 /label=L4440 /note="L4440 vector, forward primer" primer_bind 6793..6812 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" protein_bind 6941..6962 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 6977..7007 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7015..7031 /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 7020..7042 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 7039..7055 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 7039..7055 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 7105..7124 /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 taaatcgata gatcctaatc aacctctgga ttacaaaatt 3781 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct 3841 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg 3901 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc 3961 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt 4021 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc 4081 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg 4141 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg 4201 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc 4261 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg 4321 atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc 4381 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca 4441 acgaagacaa gatcacctgc aggacaggcg cgccctgctt tttgcttgta ctgggtctct 4501 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa 4561 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc 4621 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcacccgggc 4681 gattaaggaa agggctagat cattcttgaa gacgaaaggg cctcgtgata cgcctatttt 4741 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa 4801 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca 4861 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc 4921 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc 4981 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt 5041 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt 5101 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg 5161 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact 5221 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg 5281 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga 5341 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg 5401 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa 5461 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac 5521 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc 5581 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca 5641 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga 5701 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta 5761 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc 5821 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc 5881 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt 5941 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac 6001 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct 6061 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact 6121 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg 6181 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata 6241 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga 6301 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag 6361 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg 6421 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac 6481 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca 6541 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg 6601 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc 6661 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa 6721 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagcaag ctcatggctg 6781 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa 6841 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctccccg tggcacgaca 6901 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc 6961 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga 7021 gcggataaca atttcacaca ggaaacagct atgacatgat tacgaatttc acaaataaag 7081 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg 7141 tctggatcaa ctggataact caagctaacc aaaatcatcc caaacttccc accccatacc 7201 ctattaccac tgccaattac ctgtggtttc atttactcta aacctgtgat tcctctgaat 7261 tattttcatt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagt //