LOCUS       sequence-372908-        7084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7084)
  AUTHORS   Rosenbluh J, Nijhawan D, Cox AG, Li X, Neal JT, Schafer EJ, Zack TI,
            Wang X, Tsherniak A, Schinzel AC, Shao DD, Schumacher SE, Weir BA, 
            Vazquez F, Cowley GS, Root DE, Mesirov JP, Beroukhim R, Kuo CJ, 
            Goessling W, Hahn WC
  TITLE     beta-Catenin-Driven Cancers Require a YAP1 Transcriptional Complex 
            for Survival and Tumorigenesis.
  JOURNAL   Cell. 2012 Dec 21;151(7):1457-73. doi: 10.1016/j.cell.2012.11.026. 
            Epub 2012 Dec 13.
  PUBMED    23245941
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.cell.2012.11.026"; journalName: "Cell"; date: 
            "2012-12-21- 21"; volume: "151"; issue: "7"; pages: "1457-73"
COMMENT     Sequence Label: shYAP1 # 1
FEATURES             Location/Qualifiers
     source          1..7084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1932..2387)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2069..2090)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2281..2300
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2514..2536
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2528..2545
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2529..2545
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2555..2574
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2555..2573
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      2594..2729
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2769..2890
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2780..2799)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2834..2853
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             2962..3195
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     CDS             complement(3323..3922)
                     /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(3406..3426)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3903..3922
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(3944..4454)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4007..4025)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     misc_feature    4503..4620
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        complement(4717..4965)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(4775..4794)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(4945..4965)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     CDS             complement(5263..5307)
                     /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    5492..5725
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    6218..6343
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6390..6570
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(6571..6797)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(6825..6845)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6825..6843)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6864..6880)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6864..6880)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6877..6899)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6888..6904
                     /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(join(6912..6918,6919..6936,6937..6942))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6957..6978
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2461 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     2521 acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat agggcgaatt
     2581 gggtacgtcc ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc
     2641 gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg
     2701 gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggat agctagagcc
     2761 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     2821 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     2881 taaacaagtt cctctcactc tctgatattc atttctttgc aagttataaa tactgaataa
     2941 taagatgaca tgaactacta ctgctagaga ttttccacac tgactaaaag ggtctgaggg
     3001 atctctagtt accagagtca cacaacagac gggcacacac tacttgaagc actcaaggca
     3061 agctttattg aggcttaagc agtgggttcc ctagttagcc agagagctcc caggctcaga
     3121 tctggtctaa ccagagagac ccagtacaag caaaaagcag atcttgtctt cgttgggagt
     3181 gaattagccc ttccagtccc cccttttctt ttaaaaagtg gctaagatct acagctgcct
     3241 tgtaagtcat tggtcttaaa ggtaccgatg catggggtcg tgcgctcctt tcggtcgggc
     3301 gctgcgggtc gtggggcggg cgtcaggcac cgggcttgcg ggtcatgcac caggtgcgcg
     3361 gtccttcggg cacctcgacg tcggcggtga cggtgaagcc gagccgctcg tagaagggga
     3421 ggttgcgggg cgcggaggtc tccaggaagg cgggcacccc ggcgcgctcg gccgcctcca
     3481 ctccggggag cacgacggcg ctgcccagac ccttgccctg gtggtcgggc gagacgccga
     3541 cggtggccag gaaccacgcg ggctccttgg gccggtgcgg cgccaggagg ccttccatct
     3601 gttgctgcgc ggccagccgg gaaccgctca actcggccat gcgcgggccg atctcggcga
     3661 acaccgcccc cgcttcgacg ctctccggcg tggtccagac cgccaccgcg gcgccgtcgt
     3721 ccgcgaccca caccttgccg atgtcgagcc cgacgcgcgt gaggaagagt tcttgcagct
     3781 cggtgacccg ctcgatgtgg cggtccggat cgacggtgtg gcgcgtggcg gggtagtcgg
     3841 cgaacgcggc ggcgagggtg cgtacggccc tggggacgtc gtcgcgggtg gcgaggcgca
     3901 ccgtgggctt gtactcggtc atggtaagct ccggtggatc cccctgggga gagaggtcgg
     3961 tgattcggtc aacgagggag ccgactgccg acgtgcgctc cggaggcttg cagaatgcgg
     4021 aacaccgcgc gggcaggaac agggcccaca ctaccgcccc acaccccgcc tcccgcaccg
     4081 ccccttcccg gccgctgctc tcggcgcgcc ctgctgagca gccgctattg gccacagccc
     4141 atcgcggtcg gcgcgctgcc attgctccct ggcgctgtcc gtctgcgagg gtactagtga
     4201 gacgtgcggc ttccgtttgt cacgtccggc acgccgcgaa ccgcaaggaa ccttcccgac
     4261 ttaggggcgg agcaggaagc gtcgccgggg ggcccacaag ggtagcggcg aagatccggg
     4321 tgacgctgcg aacggacgtg aagaatgtgc gagacccagg gtcggcgccg ctgcgtttcc
     4381 cggaaccacg cccagagcag ccgcgtccct gcgcaaaccc agggctgcct tggaaaaggc
     4441 gcaaccccaa ccccctcgag ccgcggccaa agtggatctc tgctgtccct gtaataaacc
     4501 cgaaaatttt gaatttttgt aatttgtttt tgtaattctt tagtttgtat gtctgttgct
     4561 attatgtcta ctattctttc ccctgcactg taccccccaa tccccccttt tcttttaaaa
     4621 ttgtggatga atactgccat ttgtctcgag gtcgagaatt caaaaagcca ccaagctaga
     4681 taaagaactc gagttcttta tctagcttgg tggcccggtg tttcgtcctt tccacaagat
     4741 atataaagcc aagaaatcga aatactttca agttacggta agcatatgat agtccatttt
     4801 aaaacataat tttaaaactg caaactaccc aagaaattat tactttctac gtcacgtatt
     4861 ttgtactaat atctttgtgt ttacagtcaa attaattcca attatctctc taacagcctt
     4921 gtatcgtata tgcaaatatg aaggaatcat gggaaatagg ccctcggtga agggggcggc
     4981 cgctcgaggc tagtctcgtg atcgataccg tcgagatccg ttcactaatc gaatggatct
     5041 gtctctgtct ctctctccac cttcttcttc tattccttcg ggcctgtcgg gtcccctcgg
     5101 ggttgggagg tgggtctgaa acgataatgg tgaatatccc tgcctaactc tattcactat
     5161 agaaagtaca gcaaaaacta ttcttaaacc taccaagcct cctactatca ttatgaataa
     5221 ttttatatac cacagccaat ttgttatgtt aaaccaattc cacaaacttg cccatttatc
     5281 taattccaat aattcttgtt cattcttttc ttgctggttt tgcgattctt caattaagga
     5341 gtgtattaag cttgtgtaat tgttaatttc tctgtcccac tccatccagg tcgtgtgatt
     5401 ccaaatctgt tccagagatt tattactcca actagcattc caaggcacag cagtggtgca
     5461 aatgagtttt ccagagcaac cccaaatccc caggagctgt tgatccttta ggtatctttc
     5521 cacagccagg attcttgcct ggagctgctt gatgccccag actgtgagtt gcaacagatg
     5581 ctgttgcgcc tcaatagccc tcagcaaatt gttctgctgc tgcactatac cagacaataa
     5641 ttgtctggcc tgtaccgtca gcgtcattga cgctgcgccc atagtgcttc ctgctgctcc
     5701 caagaaccca aggaacaaag ctcctattcc cactgctctt ttttctctct gcaccactct
     5761 tctctttgcc ttggtgggtg ctactcctaa tggttcaatt tttactactt tatatttata
     5821 taattcactt ctccaattgt ccctcatatc tcctcctcca ggtctgaaga tcagcggccg
     5881 cttgctgtgc ggtggtctta cttttgtttt gctcttcctc tatcttgtct aaagcttcct
     5941 tggtgtcttt tatctctatc ctttgatgca cacaatagag ggttgctact gtattatata
     6001 atgatctaag ttcttctgat cctgtctgaa gggatggttg tagctgtccc agtatttgtc
     6061 tacagccttc tgatgtttct aacaggccag gattaactgc gaatcgttct agctccctgc
     6121 ttgcccatac tatatgtttt aatttatatt ttttctttcc ccctggcctt aaccgaattt
     6181 tttcccatcg cgatctaatt ctcccccgct taatactgac gctctcgcac ccatctctct
     6241 ccttctagcc tccgctagtc aaaatttttg gcgtactcac cagtcgccgc ccctcgcctc
     6301 ttgccgtgcg cgcttcagca agccgagtcc tgcgtcgaga gagctcctct ggtttccctt
     6361 tcgctttcaa gtccctgttc gggcgccact gctagagatt ttccacactg actaaaaggg
     6421 tctgagggat ctctagttac cagagtcaca caacagacgg gcacacacta cttgaagcac
     6481 tcaaggcaag ctttattgag gcttaagcag tgggttccct agttagccag agagctccca
     6541 ggctcagatc tggtctaacc agagagaccc gtttatgtat cgagctaggc acttaaatac
     6601 aatatctctg caatgcggca attcagtggt tcgtccaatc catgtcagac ccgtctgttg
     6661 ccttcctaat aaggcacgat cgtaccacct tacttccacc aatcggcatg cacggtgctt
     6721 tttctctcct tgtaaggcat gttgctaact catcgttacc atgttgcaag actacaagag
     6781 tattgcataa gactacatta agcttgcagc tccagctttt gttcccttta gtgagggtta
     6841 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     6901 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     6961 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     7021 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     7081 cgct
//
