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
//
