LOCUS       sequence-380256-        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 shYAP1-1
FEATURES             Location/Qualifiers
     source          1..7084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     461..481
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             692..925
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    997..1118
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1034..1053)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1088..1107
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1158..1293
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1313..1332)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1314..1332)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1342..1359)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1342..1358)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1351..1373)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1500..1955
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1587..1606)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1797..1818
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1981..2085
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2086..2154,2155..2946)
                     /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(2304..2323)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3117..3705
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3606..3625
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3859..3876
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3993..4014
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4029..4034,4035..4052,4053..4059)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4067..4083
                     /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     4072..4094
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4091..4107
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4091..4107
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4126..4146
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4128..4146
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4174..4400
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4401..4581
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4628..4753
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5246..5479
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5664..5708
                     /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        6006..6254
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6006..6026
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6177..6196
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    6351..6468
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6517..7027
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     6946..6964
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             join(7049..7084,1..564)
                     /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(7049..7068)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
       61 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
      121 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
      181 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
      241 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
      301 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
      361 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
      421 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
      481 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
      541 atgacccgca agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg
      601 agcgcacgac cccatgcatc ggtaccttta agaccaatga cttacaaggc agctgtagat
      661 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga
      721 caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag
      781 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
      841 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
      901 tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta
      961 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat
     1021 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     1081 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc
     1141 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     1201 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     1261 tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag
     1321 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     1381 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     1441 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     1501 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     1561 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     1621 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     1681 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     1741 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     1801 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     1861 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     1921 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     1981 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2041 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     2101 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     2161 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     2221 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     2281 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     2341 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     2401 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     2461 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     2521 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     2581 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     2641 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     2701 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     2761 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     2821 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     2881 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     2941 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     3001 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     3061 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3121 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3181 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3241 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     3301 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3361 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3421 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3481 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3541 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3601 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3661 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3721 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     3781 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     3841 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     3901 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     3961 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     4021 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     4081 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac
     4141 taaagggaac aaaagctgga gctgcaagct taatgtagtc ttatgcaata ctcttgtagt
     4201 cttgcaacat ggtaacgatg agttagcaac atgccttaca aggagagaaa aagcaccgtg
     4261 catgccgatt ggtggaagta aggtggtacg atcgtgcctt attaggaagg caacagacgg
     4321 gtctgacatg gattggacga accactgaat tgccgcattg cagagatatt gtatttaagt
     4381 gcctagctcg atacataaac gggtctctct ggttagacca gatctgagcc tgggagctct
     4441 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     4501 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     4561 cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc
     4621 agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg
     4681 gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg
     4741 gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt
     4801 aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct
     4861 agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact
     4921 gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac
     4981 agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt
     5041 agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga
     5101 tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata
     5161 aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc
     5221 agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag
     5281 gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg
     5341 gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc
     5401 aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc
     5461 taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg
     5521 ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga
     5581 cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg
     5641 aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg
     5701 caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa
     5761 tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata
     5821 gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac
     5881 ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc
     5941 gattagtgaa cggatctcga cggtatcgat cacgagacta gcctcgagcg gccgccccct
     6001 tcaccgaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta
     6061 gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
     6121 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
     6181 atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga
     6241 aaggacgaaa caccggccca gttaaatgtt caccaatctc gagattggtg aacatttaac
     6301 tgggtttttg aattctcgac ctcgagacaa atggcagtat tcatccacaa ttttaaaaga
     6361 aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac
     6421 atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac
     6481 agggacagca gagatccact ttggccgcgg ctcgaggggg ttggggttgc gccttttcca
     6541 aggcagccct gggtttgcgc agggacgcgg ctgctctggg cgtggttccg ggaaacgcag
     6601 cggcgccgac cctgggtctc gcacattctt cacgtccgtt cgcagcgtca cccggatctt
     6661 cgccgctacc cttgtgggcc ccccggcgac gcttcctgct ccgcccctaa gtcgggaagg
     6721 ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa gccgcacgtc tcactagtac
     6781 cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc cgaccgcgat gggctgtggc
     6841 caatagcggc tgctcagcag ggcgcgccga gagcagcggc cgggaagggg cggtgcggga
     6901 ggcggggtgt ggggcggtag tgtgggccct gttcctgccc gcgcggtgtt ccgcattctg
     6961 caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt gaccgaatca ccgacctctc
     7021 tccccagggg gatccaccgg agcttaccat gaccgagtac aagcccacgg tgcgcctcgc
     7081 cacc
//
