LOCUS Exported 8477 bp ds-DNA circular SYN 06-JAN-2022 DEFINITION doxycycline-inducible lentiviral vector. ACCESSION . VERSION . KEYWORDS FUW-tetO-MCS SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8477) AUTHORS Panciera T, Azzolin L, Fujimura A, Di Biagio D, Frasson C, Bresolin S, Soligo S, Basso G, Bicciato S, Rosato A, Cordenonsi M, Piccolo S TITLE Induction of Expandable Tissue-Specific Stem/Progenitor Cells through Transient Expression of YAP/TAZ. JOURNAL Cell Stem Cell. 2016 Sep 9. pii: S1934-5909(16)30256-9. doi: 10.1016/j.stem.2016.08.009. PUBMED 27641305 REFERENCE 2 (bases 1 to 8477) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 6, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8477 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 27..406 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 408..606 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 561..581 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 624..804 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 851..976 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1469..1702 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1887..1931 /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 2229..2346 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" protein_bind 2399..2669 /label=tetracycline response element /bound_moiety="tetracycline repressor TetR" /note="contains seven copies of the tetracycline operator tetO" promoter 2702..2740 /label=minimal CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2737..2761 /label=LNCX /note="Human CMV promoter, forward primer" misc_feature 2958..3546 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3011..3031) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3429..3440) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(3549..3565) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3550..3566) /label=pBluescriptKS /note="For pBluescript vector" LTR 4071..4251 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(4277..4294) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4283..4507 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 4553..4981 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4640..4659) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4850..4871 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4990..5010) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4995..5324 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 5175..5310 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5237..5256 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 5372..5419 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 5438..5812 /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" polyA_signal 5942..6063 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5979..5998) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6033..6052 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(6112..6128) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6112..6128) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6125..6147) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6136..6152 /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)." promoter complement(6160..6190) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6205..6226 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6343..6360) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6514..7102) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6594..6613) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7273..8133) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 7896..7915 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8134..8238) /gene="bla" /label=AmpR promoter primer_bind complement(8313..8332) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc 61 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt 121 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta 181 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg 241 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga 301 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt 361 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg 421 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc 481 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg 541 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat 601 aagcagcgcg ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc 661 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc 721 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt 781 agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa 841 accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga 901 ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga 961 tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg 1021 gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga 1081 gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat 1141 actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa 1201 tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc 1261 tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc 1321 tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat 1381 ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg 1441 tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag 1501 caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt 1561 ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt 1621 tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat 1681 acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca 1741 ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca 1801 cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa 1861 ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat 1921 gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata aaattattca 1981 taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga 2041 atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg 2101 gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca 2161 ttcgattagt gaacggatcg gcactgcgtg cgccaattct gcagacaaat ggcagtattc 2221 atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta 2281 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa 2341 aattttcggg tttattacag ggacagcaga gatccagttt ggttatcgag tttaccactc 2401 cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa 2461 aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta 2521 ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat 2581 agagaaaagt gaaagtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg 2641 agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagctcggt acccgggtcg 2701 aggtaggcgt gtacggtggg aggcctatat aagcagagct cgtttagtga accgtcagat 2761 cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg accgatccag 2821 cctccgcggc cccgaattac aggctagcta tcaggatcct acgaggcgcg ccagatagaa 2881 ttctcttggg cctcaagggc catgagttaa ccagttctag atgtgactca tgtcaattcg 2941 atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta 3001 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc 3061 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt 3121 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg 3181 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt 3241 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct 3301 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt 3361 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct 3421 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc 3481 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct 3541 ccccgcatcg ataccgtcga cctcgagacc tagaaaaaca tggagcaatc acaagtagca 3601 atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg 3661 gttttccagt cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc 3721 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac 3781 aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat tggcagaact 3841 acacaccagg gccagggatc agatatccac tgacctttgg atggtgctac aagctagtac 3901 cagttgagca agagaaggta gaagaagcca atgaaggaga gaacacccgc ttgttacacc 3961 ctgtgagcct gcatgggatg gatgacccgg agagagaagt attagagtgg aggtttgaca 4021 gccgcctagc atttcatcac atggcccgag agctgcatcc ggactgtact gggtctctct 4081 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc 4141 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg 4201 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agggcccgtt 4261 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc 4321 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat 4381 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg 4441 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc 4501 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc 4561 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt 4621 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc 4681 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta 4741 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc 4801 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg 4861 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt 4921 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat 4981 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca 5041 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct 5101 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc 5161 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg 5221 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc 5281 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt 5341 gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat 5401 cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt 5461 tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg 5521 gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct 5581 gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt 5641 gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga 5701 cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct 5761 gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtgct 5821 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg 5881 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc 5941 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 6001 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 6061 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct 6121 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat 6181 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc 6241 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 6301 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 6361 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 6421 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 6481 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 6541 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 6601 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 6661 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 6721 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 6781 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 6841 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 6901 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt 6961 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 7021 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 7081 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 7141 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 7201 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 7261 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 7321 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 7381 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 7441 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 7501 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 7561 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 7621 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 7681 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 7741 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 7801 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 7861 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 7921 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 7981 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 8041 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 8101 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 8161 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 8221 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg 8281 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta 8341 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt 8401 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg 8461 cgttttgcgc tgcttcg //