LOCUS Exported 7216 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Dox-inducible lentiviral vector expressing mouse Sox2. ACCESSION . VERSION . KEYWORDS pHAGE2-TetOminiCMV-Sox2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7216) 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 7216) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7216 /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" misc_feature 3658..4246 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3711..3731) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4129..4140) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 4394..4574 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(4621..4639) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4707..4811 /gene="bla" /label=AmpR promoter CDS 4812..5672 /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(5030..5049) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5843..6431 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6332..6351 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6585..6602 /label=L4440 /note="L4440 vector, forward primer" primer_bind 6694..6713 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" protein_bind 6842..6863 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 6878..6908 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6916..6932 /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 6921..6943 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 6940..6956 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 6940..6956 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 7006..7025 /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 gtaaatcgat agatcctaat 3661 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct 3721 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg 3781 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg 3841 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt 3901 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt 3961 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg 4021 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc 4081 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat 4141 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc 4201 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg tacctttaag 4261 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact 4321 ggaagggcta attcactccc aacgaagaca agatcacctg caggacaggc gcgccctgct 4381 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta 4441 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg 4501 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg 4561 gaaaatctct agcacccggg cgattaagga aagggctaga tcattcttga agacgaaagg 4621 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt 4681 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac 4741 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa 4801 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat 4861 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc 4921 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga 4981 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg 5041 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc 5101 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag 5161 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc 5221 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg 5281 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg 5341 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac 5401 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac 5461 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg 5521 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg 5581 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg 5641 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac 5701 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg 5761 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 5821 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 5881 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 5941 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt 6001 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 6061 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 6121 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 6181 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 6241 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 6301 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 6361 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 6421 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 6481 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct 6541 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg 6601 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt 6661 aatgcagcaa gctcatggct gactaatttt ttttatttat gcagaggccg aggccgcctc 6721 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa 6781 aaagctcccc gtggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa 6841 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc 6901 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgacatga 6961 ttacgaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac 7021 tcatcaatgt atcttatcat gtctggatca actggataac tcaagctaac caaaatcatc 7081 ccaaacttcc caccccatac cctattacca ctgccaatta cctgtggttt catttactct 7141 aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt taaatatgta 7201 ctacaaactt agtagt //