LOCUS       sequence-380254-        7084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive lentiviral expression of YAP1 shRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7084)
  AUTHORS   Kim M, Ly SH, Xie Y, Duronio GN, Ford-Roshon D, Hwang JH, Sulahian 
            R, Rennhack JP, So J, Gjoerup O, Talamas JA, Grandclaudon M, Long 
            HW, Doench JG, Sethi NS, Giannakis M, Hahn WC
  TITLE     YAP1 and PRDM14 converge to promote cell survival and tumorigenesis.
  JOURNAL   Dev Cell. 2022 Jan 24;57(2):212-227.e8. doi: 
            10.1016/j.devcel.2021.12.006. Epub 2022 Jan 5.
  PUBMED    34990589
REFERENCE   2  (bases 1 to 7084)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.devcel.2021.12.006"; journalName: "Dev Cell"; date: 
            "2022-01-24- 24"; volume: "57"; issue: "2"; pages: "212-227.e8"
COMMENT     Sequence Label: pLKO.1 puro shYAP1-2
FEATURES             Location/Qualifiers
     source          1..7084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     449..469
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             680..913
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    985..1106
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1022..1041)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1076..1095
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1146..1281
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1301..1320)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1302..1320)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1330..1347)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1330..1346)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1339..1361)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1488..1943
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1575..1594)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1785..1806
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1969..2073
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2074..2142,2143..2934)
                     /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     complement(2292..2311)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3105..3693
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3594..3613
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3847..3864
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3981..4002
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4017..4022,4023..4040,4041..4047)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4055..4071
                     /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     4060..4082
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4079..4095
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4079..4095
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4114..4134
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4116..4134
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4162..4388
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4389..4569
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4616..4741
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5234..5467
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5652..5696
                     /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"
     promoter        5994..6242
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5994..6014
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6165..6184
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    6339..6456
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6505..7015
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     6934..6952
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             join(7037..7084,1..552)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(7037..7056)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
       61 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
      121 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
      181 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
      241 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
      301 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
      361 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
      421 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
      481 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
      541 cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc
      601 catgcatcgg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt
      661 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt
      721 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
      781 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
      841 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
      901 aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag
      961 aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa
     1021 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     1081 ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc
     1141 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     1201 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     1261 gaggaggctt ttttggaggc ctagggacgt acccaattcg ccctatagtg agtcgtatta
     1321 cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     1381 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     1441 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag
     1501 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     1561 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     1621 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     1681 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     1741 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     1801 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     1861 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     1921 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
     1981 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2041 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     2101 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     2161 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     2221 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     2281 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     2341 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     2401 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     2461 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     2521 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     2581 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     2641 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     2701 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     2761 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     2821 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     2881 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     2941 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     3001 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     3061 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     3121 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3181 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     3241 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     3301 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     3361 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     3421 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     3481 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     3541 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     3601 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     3661 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     3721 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     3781 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     3841 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     3901 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     3961 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     4021 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     4081 ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa
     4141 aagctggagc tgcaagctta atgtagtctt atgcaatact cttgtagtct tgcaacatgg
     4201 taacgatgag ttagcaacat gccttacaag gagagaaaaa gcaccgtgca tgccgattgg
     4261 tggaagtaag gtggtacgat cgtgccttat taggaaggca acagacgggt ctgacatgga
     4321 ttggacgaac cactgaattg ccgcattgca gagatattgt atttaagtgc ctagctcgat
     4381 acataaacgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     4441 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     4501 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     4561 tctctagcag tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc
     4621 tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt
     4681 gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt
     4741 cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg
     4801 aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc
     4861 agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca
     4921 accatccctt cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct
     4981 ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag acaagataga
     5041 ggaagagcaa aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg
     5101 gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa
     5161 ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa
     5221 gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg
     5281 gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc
     5341 agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct
     5401 ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac
     5461 agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga
     5521 atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt
     5581 gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa
     5641 accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga
     5701 attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag
     5761 gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg
     5821 gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg
     5881 aaggaataga agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg
     5941 gatctcgacg gtatcgatca cgagactagc ctcgagcggc cgcccccttc accgagggcc
     6001 tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga gagataattg
     6061 gaattaattt gactgtaaac acaaagatat tagtacaaaa tacgtgacgt agaaagtaat
     6121 aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat catatgctta
     6181 ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttgtggaaa ggacgaaaca
     6241 ccgggccacc aagctagata aagaactcga gttctttatc tagcttggtg gctttttgaa
     6301 ttctcgacct cgagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt
     6361 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
     6421 gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga
     6481 gatccacttt ggccgcggct cgagggggtt ggggttgcgc cttttccaag gcagccctgg
     6541 gtttgcgcag ggacgcggct gctctgggcg tggttccggg aaacgcagcg gcgccgaccc
     6601 tgggtctcgc acattcttca cgtccgttcg cagcgtcacc cggatcttcg ccgctaccct
     6661 tgtgggcccc ccggcgacgc ttcctgctcc gcccctaagt cgggaaggtt ccttgcggtt
     6721 cgcggcgtgc cggacgtgac aaacggaagc cgcacgtctc actagtaccc tcgcagacgg
     6781 acagcgccag ggagcaatgg cagcgcgccg accgcgatgg gctgtggcca atagcggctg
     6841 ctcagcaggg cgcgccgaga gcagcggccg ggaaggggcg gtgcgggagg cggggtgtgg
     6901 ggcggtagtg tgggccctgt tcctgcccgc gcggtgttcc gcattctgca agcctccgga
     6961 gcgcacgtcg gcagtcggct ccctcgttga ccgaatcacc gacctctctc cccaggggga
     7021 tccaccggag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     7081 cgtc
//
