LOCUS       sequence-336441-        7082 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 7082)
  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 7082)
  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: shCTNNB1 # 2
FEATURES             Location/Qualifiers
     source          1..7082
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     128..147
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     381..398
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    515..536
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(551..556,557..574,575..581)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    589..605
                     /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     594..616
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     613..629
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     613..629
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     648..668
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        650..668
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        696..922
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             923..1103
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1150..1275
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1768..2001
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2186..2230
                     /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        2528..2776
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2528..2548
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2699..2718
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    2872..2989
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3038..3548
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3467..3485
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             3570..4169
                     /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(3570..3589)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4066..4086
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4297..4530
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4602..4723
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4639..4658)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4693..4712
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4763..4898
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4918..4937)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4919..4937)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4947..4964)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4947..4963)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4956..4978)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5105..5560
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5192..5211)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5402..5423
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5586..5690
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5691..5759,5760..6551)
                     /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(5909..5928)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6722..7082,1..227)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
       61 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
      121 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
      181 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
      241 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
      301 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
      361 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
      421 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
      481 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
      541 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
      601 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
      661 actaaaggga acaaaagctg gagctgcaag cttaatgtag tcttatgcaa tactcttgta
      721 gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga aaaagcaccg
      781 tgcatgccga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa ggcaacagac
      841 gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata ttgtatttaa
      901 gtgcctagct cgatacataa acgggtctct ctggttagac cagatctgag cctgggagct
      961 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     1021 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     1081 gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa
     1141 ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag
     1201 gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat
     1261 gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg
     1321 ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag
     1381 ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata
     1441 ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat
     1501 acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct
     1561 ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct
     1621 gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata
     1681 taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt
     1741 gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc
     1801 aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc
     1861 tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt
     1921 gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata
     1981 cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac
     2041 tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac
     2101 gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat
     2161 tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg
     2221 ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa aattattcat
     2281 aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa
     2341 tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg
     2401 acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag acagatccat
     2461 tcgattagtg aacggatctc gacggtatcg atcacgagac tagcctcgag cggccgcccc
     2521 cttcaccgag ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt
     2581 tagagagata attggaatta atttgactgt aaacacaaag atattagtac aaaatacgtg
     2641 acgtagaaag taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga
     2701 ctatcatatg cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg
     2761 gaaaggacga aacaccgggc ttggaatgag actgctgatc tcgagatcag cagtctcatt
     2821 ccaagctttt taattctcga cctcgagaca aatggcagta ttcatccaca attttaaaag
     2881 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
     2941 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta
     3001 cagggacagc agagatccac tttggccgcg gctcgagggg gttggggttg cgccttttcc
     3061 aaggcagccc tgggtttgcg cagggacgcg gctgctctgg gcgtggttcc gggaaacgca
     3121 gcggcgccga ccctgggtct cgcacattct tcacgtccgt tcgcagcgtc acccggatct
     3181 tcgccgctac ccttgtgggc cccccggcga cgcttcctgc tccgccccta agtcgggaag
     3241 gttccttgcg gttcgcggcg tgccggacgt gacaaacgga agccgcacgt ctcactagta
     3301 ccctcgcaga cggacagcgc cagggagcaa tggcagcgcg ccgaccgcga tgggctgtgg
     3361 ccaatagcgg ctgctcagca gggcgcgccg agagcagcgg ccgggaaggg gcggtgcggg
     3421 aggcggggtg tggggcggta gtgtgggccc tgttcctgcc cgcgcggtgt tccgcattct
     3481 gcaagcctcc ggagcgcacg tcggcagtcg gctccctcgt tgaccgaatc accgacctct
     3541 ctccccaggg ggatccaccg gagcttacca tgaccgagta caagcccacg gtgcgcctcg
     3601 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     3661 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     3721 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     3781 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     3841 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     3901 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     3961 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     4021 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     4081 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     4141 ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg
     4201 aaaggagcgc acgaccccat gcatcggtac ctttaagacc aatgacttac aaggcagctg
     4261 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     4321 gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     4381 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     4441 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     4501 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
     4561 atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt
     4621 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4681 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc
     4741 tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4801 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     4861 gctattccag aagtagtgag gaggcttttt tggaggccta gggacgtacc caattcgccc
     4921 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa
     4981 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     5041 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     5101 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     5161 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     5221 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     5281 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     5341 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     5401 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     5461 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     5521 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
     5581 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5641 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     5701 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     5761 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     5821 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     5881 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5941 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     6001 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     6061 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     6121 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     6181 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     6241 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     6301 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     6361 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     6421 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     6481 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     6541 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     6601 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6661 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6721 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6781 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6841 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     6901 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6961 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     7021 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     7081 cc
//
