LOCUS       sequence-336439-        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: shCTNNB1 # 1
FEATURES             Location/Qualifiers
     source          1..7084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    90..111
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(126..131,132..149,150..156)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    164..180
                     /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     169..191
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     188..204
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     188..204
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     223..243
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        225..243
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        271..497
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             498..678
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    725..850
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1343..1576
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1761..1805
                     /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        2103..2351
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2103..2123
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2274..2293
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    2448..2565
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2614..3124
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3043..3061
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             3146..3745
                     /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(3146..3165)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3642..3662
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3873..4106
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4178..4299
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4215..4234)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4269..4288
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4339..4474
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4494..4513)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4495..4513)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4523..4540)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4523..4539)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4532..4554)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4681..5136
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4768..4787)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4978..4999
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5162..5266
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5267..5335,5336..6127)
                     /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(5485..5504)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6298..6886
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6787..6806
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7040..7057
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
       61 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
      121 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
      181 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
      241 agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc ttgtagtctt
      301 gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag caccgtgcat
      361 gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa cagacgggtc
      421 tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta tttaagtgcc
      481 tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg gagctctctg
      541 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
      601 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
      661 tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga
      721 ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg
      781 gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg
      841 cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag
      901 gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga
      961 acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg
     1021 acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt
     1081 agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga
     1141 caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct
     1201 tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat aaatataaag
     1261 tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga
     1321 gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa
     1381 gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta
     1441 tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac
     1501 tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa
     1561 aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg
     1621 tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct
     1681 ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag
     1741 aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa
     1801 gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga
     1861 tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag
     1921 ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg aggggacccg
     1981 acaggcccga aggaatagaa gaagaaggtg gagagagaga cagagacaga tccattcgat
     2041 tagtgaacgg atctcgacgg tatcgatcac gagactagcc tcgagcggcc gcccccttca
     2101 ccgagggcct atttcccatg attccttcat atttgcatat acgatacaag gctgttagag
     2161 agataattgg aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta
     2221 gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc
     2281 atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag
     2341 gacgaaacac cggggtgtct gctattgtac gtacctcgag gtacgtacaa tagcagacac
     2401 ctttttgaat tctcgacctc gagacaaatg gcagtattca tccacaattt taaaagaaaa
     2461 ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata
     2521 caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg
     2581 gacagcagag atccactttg gccgcggctc gagggggttg gggttgcgcc ttttccaagg
     2641 cagccctggg tttgcgcagg gacgcggctg ctctgggcgt ggttccggga aacgcagcgg
     2701 cgccgaccct gggtctcgca cattcttcac gtccgttcgc agcgtcaccc ggatcttcgc
     2761 cgctaccctt gtgggccccc cggcgacgct tcctgctccg cccctaagtc gggaaggttc
     2821 cttgcggttc gcggcgtgcc ggacgtgaca aacggaagcc gcacgtctca ctagtaccct
     2881 cgcagacgga cagcgccagg gagcaatggc agcgcgccga ccgcgatggg ctgtggccaa
     2941 tagcggctgc tcagcagggc gcgccgagag cagcggccgg gaaggggcgg tgcgggaggc
     3001 ggggtgtggg gcggtagtgt gggccctgtt cctgcccgcg cggtgttccg cattctgcaa
     3061 gcctccggag cgcacgtcgg cagtcggctc cctcgttgac cgaatcaccg acctctctcc
     3121 ccagggggat ccaccggagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac
     3181 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     3241 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     3301 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     3361 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     3421 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     3481 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     3541 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     3601 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     3661 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     3721 catgacccgc aagcccggtg cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag
     3781 gagcgcacga ccccatgcat cggtaccttt aagaccaatg acttacaagg cagctgtaga
     3841 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
     3901 acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
     3961 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     4021 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     4081 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt
     4141 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa
     4201 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4261 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc
     4321 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4381 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     4441 ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata
     4501 gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     4561 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     4621 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg
     4681 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4741 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4801 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4861 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     4921 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4981 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     5041 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     5101 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
     5161 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     5221 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     5281 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5341 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     5401 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     5461 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     5521 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5581 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5641 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     5701 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     5761 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     5821 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     5881 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5941 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     6001 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     6061 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     6121 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     6181 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6241 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6301 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6361 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6421 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6481 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6541 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6601 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6661 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6721 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6781 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6841 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6901 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6961 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     7021 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     7081 caaa
//
