LOCUS       sequence-381444-        3332 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Carries a Sleeping Beauty (SB) transposon vector that can be used to
            deliver an activated human CTNNB1(S33Y) transgene into cells, when a
            source of SB transposase is also co-introduced..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3332)
  AUTHORS   Keng VW, Sia D, Sarver AL, Tschida BR, Fan D, Alsinet C, Sole M, Lee
            WL, Kuka TP, Moriarity BS, Villanueva A, Dupuy AJ, Riordan JD, Bell 
            JB, Silverstein KA, Llovet JM, Largaespada DA
  TITLE     Sex bias occurrence of hepatocellular carcinoma in Poly7 molecular 
            subclass is associated with EGFR.
  JOURNAL   Hepatology. 2013 Jan;57(1):120-30. doi: 10.1002/hep.26004. Epub 2012
            Oct 19.
  PUBMED    22899566
REFERENCE   2  (bases 1 to 3332)
  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: "10.1002/hep.26004";
            journalName: "Hepatology"; date: "2013-01"; volume: "57"; issue: 
            "1"; pages: "120-30"
COMMENT     Sequence Label: pT2/GD-IRES-GFP-CTNNB1
FEATURES             Location/Qualifiers
     source          1..3332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(3..20)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(174..762)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(254..273)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(933..1724,1725..1793))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAAKLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1556..1575
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1794..1898)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1925..2380
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2012..2031)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2222..2243
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2506..2528
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2520..2537
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2521..2537
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2547..2566
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2547..2565
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3065..3085)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3065..3083)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3104..3120)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3104..3120)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3117..3139)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3128..3144
                     /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(3152..3158,3159..3176,3177..3182))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3197..3218
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
       61 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      121 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      181 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      241 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      301 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aaccgtggcg
      361 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      421 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      481 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      541 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      601 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      661 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      721 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      781 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      841 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
      901 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
      961 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1021 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1081 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1141 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1201 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1261 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1321 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1381 gtcagaagta acttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1441 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1501 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     1561 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1621 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1681 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1741 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     1801 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1861 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     1921 cctgacgcgc cctgtagcgg cgcattaagc gcggcgggag tggtggttac gcgcagcgtg
     1981 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2041 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2101 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     2161 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2221 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     2281 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2341 tttaacgcga attttaacaa aatattaacg cttacaattt ccattcgcca ttcaggctgc
     2401 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     2461 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     2521 gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttggagctcg
     2581 gatccctata cagttgaagt cggaagttta catacactta agttggagtc attaaaactc
     2641 gtttttcaac tactcccagt catagctgtc cctcttctct tatggagatc cctcgacctg
     2701 cataccggtc aagctaggct actcgaaatg tttgacccaa gttaaacaat ttaaaggcaa
     2761 tgctaccaaa tactaattga gtgtatgtaa acttctgacc cactgggaat gtgatgaaag
     2821 aaataaaagc tgaaatgaat cattctctct actattattc tgatatttca cattcttaaa
     2881 ataaagtggt gatcctaact gacctaagac agggaatttt tactaggatt aaatgtcagg
     2941 aattgtgaaa aagtgagttt aaatgtattt ggctaaggtg tatgtaaact tccgacttca
     3001 actgtatagg gatcctctag ctagagtcga cctcgagggg gggcccggta cccagctttt
     3061 gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg
     3121 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3181 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     3241 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3301 gaggcggttt gcgtattggg cgctcttccg ct
//
