LOCUS       V000016                 8815 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V000016
VERSION     V000016
KEYWORDS    Tet-pLKO-puro-Scrambled
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8815)
  AUTHORS   Rudin CM, Durinck S, Stawiski EW, Poirier JT, Modrusan Z, Shames DS,
            Bergbower EA, Guan Y, Shin J, Guillory J, Rivers CS, Foo CK, Bhatt
            D, Stinson J, Gnad F, Haverty PM, Gentleman R, Chaudhuri S,
            Janakiraman V, Jaiswal BS, Parikh C, Yuan W, Zhang Z, Koeppen H, Wu
            TD, Stern HM, Yauch RL, Huffman KE, Paskulin DD, Illei PB,
            Varella-Garcia M, Gazdar AF, de Sauvage FJ, Bourgon R, Minna JD,
            Brock MV, Seshagiri S
  TITLE     Comprehensive genomic analysis identifies SOX2 as a frequently
            amplified gene in small-cell lung cancer.
  JOURNAL   Nat Genet. 2012 Oct;44(10):1111-6. doi: 10.1038/ng.2405. Epub 2012
            Sep 2.
   PUBMED   22941189
REFERENCE   2  (bases 1 to 8815)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8815)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1038/ng.2405";
            journalName: "Nat Genet"; date: "2012-10"; volume: "44"; issue:
            "10"; pages: "1111-6"
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8815
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             489..722
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    794..928
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      955..1090
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(1111..1129)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1139..1155)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      1297..1752
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1778..1882
                     /label="AmpR promoter"
     CDS             1883..2740
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      2914..3502
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    3790..3811
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3826..3856
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    3864..3880
                     /label="lac operator"
                     /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     3888..3904
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        3925..3943
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3971..4197
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     misc_feature    4425..4550
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    5043..5276
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             5461..5505
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     CDS             5654..5695
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     promoter        5794..5888
                     /label="H1-2O2 promoter"
                     /note="doxycycline-inducible variant of the human H1 RNA
                     promoter (Henriksen et al., 2007)"
     misc_feature    5988..6105
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        6154..6664
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     intron          6673..7245
                     /label="beta-globin intron"
                     /note="intron from rabbit beta-globin gene"
     promoter        7300..7318
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7329..7949
                     /label="TetR"
                     /note="tetracycline repressor TetR"
     misc_feature    7984..8557
                     /label="IRES"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     CDS             join(8577..8815,1..358)
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
ORIGIN
        1 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
       61 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
      121 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
      181 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
      241 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
      301 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
      361 acgcccgccc cacgacccgc agcgcccgac cgaaaggagc gcacgacccc atgcatcggt
      421 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
      481 ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac
      541 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
      601 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
      661 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
      721 cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc
      781 agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
      841 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
      901 catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccatcccg
      961 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     1021 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     1081 tttggaggcc tagggacgta cccaattcgc cctatagtga gtcgtattac gcgcgctcac
     1141 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     1201 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     1261 cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa
     1321 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     1381 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     1441 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     1501 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     1561 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     1621 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     1681 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     1741 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     1801 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     1861 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     1921 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     1981 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     2041 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     2101 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     2161 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     2221 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     2281 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     2341 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     2401 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     2461 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     2521 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     2581 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     2641 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     2701 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     2761 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     2821 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     2881 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2941 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3001 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3061 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3121 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3181 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3241 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3301 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3361 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3421 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3481 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3541 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     3601 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     3661 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     3721 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     3781 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     3841 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     3901 tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa agctggagct
     3961 gcaagcttaa tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt
     4021 tagcaacatg ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg
     4081 tggtacgatc gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc
     4141 actgaattgc cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg
     4201 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     4261 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     4321 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     4381 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
     4441 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
     4501 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
     4561 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     4621 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     4681 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     4741 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     4801 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     4861 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     4921 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     4981 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     5041 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     5101 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     5161 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     5221 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     5281 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     5341 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     5401 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     5461 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     5521 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     5581 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     5641 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     5701 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg
     5761 tatcgatcac gagactagcc tcgagcggcc gcaatatttg catgtcgcta tgtgttctgg
     5821 gaaatcacca taaacgtgaa atccctatca gtgatagaga cttataagtt ccctatcagt
     5881 gatagagaca ccggcctaag gttaagtcgc cctcgctcga gcgagggcga cttaacctta
     5941 ggtttttaat tctcgacctc gagacaaatg gcagtattca tccacaattt taaaagaaaa
     6001 ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata
     6061 caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg
     6121 gacagcagag atccactttg gccgcggctc gagggggttg gggttgcgcc ttttccaagg
     6181 cagccctggg tttgcgcagg gacgcggctg ctctgggcgt ggttccggga aacgcagcgg
     6241 cgccgaccct gggtctcgca cattcttcac gtccgttcgc agcgtcaccc ggatcttcgc
     6301 cgctaccctt gtgggccccc cggcgacgct tcctgctccg cccctaagtc gggaaggttc
     6361 cttgcggttc gcggcgtgcc ggacgtgaca aacggaagcc gcacgtctca ctagtaccct
     6421 cgcagacgga cagcgccagg gagcaatggc agcgcgccga ccgcgatggg ctgtggccaa
     6481 tagcggctgc tcagcagggc gcgccgagag cagcggccgg gaaggggcgg tgcgggaggc
     6541 ggggtgtggg gcggtagtgt gggccctgtt cctgcccgcg cggtgttccg cattctgcaa
     6601 gcctccggag cgcacgtcgg cagtcggctc cctcgttgac cgaatcaccg acctctctcc
     6661 ccagggggat ctgtgagttt ggggaccctt gattgttctt tctttttcgc tattgtaaaa
     6721 ttcatgttat atggaggggg caaagttttc agggtgttgt ttagaatggg aagatgtccc
     6781 ttgtatcacc atggaccctc atgataattt tgtttctttc actttctact ctgttgacaa
     6841 ccattgtctc ctcttatttt cttttcattt tctgtaactt tttcgttaaa ctttagcttg
     6901 catttgtaac gaatttttaa attcactttt gtttatttgt cagattgtaa gtactttctc
     6961 taatcacttt tttttcaagg caatcagggt atattatatt gtacttcagc acagttttag
     7021 agaacaattg ttataattaa atgataaggt agaatatttc tgcatataaa ttctggctgg
     7081 cgtggaaata ttcttattgg tagaaacaac tacatcctgg tcatcatcct gcctttctct
     7141 ttatggttac aatgatatac actgtttgag atgaggataa aatactctga gtccaaaccg
     7201 ggcccctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg
     7261 ctggttattg tgctgtctca tcattttggc aaagaattgt aatacgactc actatagggc
     7321 gaattgatat gtctagatta gataaaagta aagtgattaa cagcgcatta gagctgctta
     7381 atgaggtcgg aatcgaaggt ttaacaaccc gtaaactcgc ccagaagcta ggtgtagagc
     7441 agcctacatt gtattggcat gtaaaaaata agcgggcttt gctcgacgcc ttagccattg
     7501 agatgttaga taggcaccat actcactttt gccctttaga aggggaaagc tggcaagatt
     7561 ttttacgtaa taacgctaaa agttttagat gtgctttact aagtcatcgc gatggagcaa
     7621 aagtacattt aggtacacgg cctacagaaa aacagtatga aactctcgaa aatcaattag
     7681 cctttttatg ccaacaaggt ttttcactag agaatgcatt atatgcactc agcgctgtgg
     7741 ggcattttac tttaggttgc gtattggaag atcaagagca tcaagtcgct aaagaagaaa
     7801 gggaaacacc tactactgat agtatgccgc cattattacg acaagctatc gaattatttg
     7861 atcaccaagg tgcagagcca gccttcttat tcggccttga attgatcata tgcggattag
     7921 aaaaacaact taaatgtgaa agtgggtccg cgtacagcgg ctcccgggag ttctagggat
     7981 ctgcccctct ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg
     8041 gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc
     8101 ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa
     8161 aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag
     8221 acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg
     8281 cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg
     8341 ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa
     8401 caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg
     8461 gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca
     8521 cggggacgtg gttttccttt gaaaaacacg atgataagga tccaccggag cttaccatga
     8581 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca
     8641 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc
     8701 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     8761 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccgga
//
