LOCUS       V024916                 7780 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024916
VERSION     V024916
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        1..1179
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
                     /label="EF-1α promoter"
     promoter        1196..1214
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     protein_bind    1293..1317
                     /bound_moiety="BP Clonase™"
                     /gene="mutant version of&nbsp;attB"
                     /note="recombination site for the Gateway® BP reaction"
                     /label="attB1"
     CDS             1338..3683
                     /codon_start=1
                     /gene="CTNNB1"
                     /note="also known as catenin beta-1; related to Drosophila
                     armadillo"
                     /product="protein involved both in cell-cell adhesion as
                     part of the cadherin protein complex, and in gene
                     transcription as part of the Wnt signaling pathway"
                     /protein_id=""
                     /transl_table=1
                     /translation="MATQADLMELDMAMEPDRKAAVSHWQQQSYLDSGIHSGATTTAPS
                     LSGKGNPEEEDVDTSQVLYEWEQGFSQSFTQEQVADIDGQYAMTRAQRVRAAMFPETLD
                     EGMQIPSTQFDAAHPTNVQRLAEPSQMLKHAVVNLINYQDDAELATRAIPELTKLLNDE
                     DQVVVNKAAVMVHQLSKKEASRHAIMRSPQMVSAIVRTMQNTNDVETARCTAGTLHNLS
                     HHREGLLAIFKSGGIPALVKMLGSPVDSVLFYAITTLHNLLLHQEGAKMAVRLAGGLQK
                     MVALLNKTNVKFLAITTDCLQILAYGNQESKLIILASGGPQALVNIMRTYTYEKLLWTT
                     SRVLKVLSVCSSNKPAIVEAGGMQALGLHLTDPSQRLVQNCLWTLRNLSDAATKQEGME
                     GLLGTLVQLLGSDDINVVTCAAGILSNLTCNNYKNKMMVCQVGGIEALVRTVLRAGDRE
                     DITEPAICALRHLTSRHQEAEMAQNAVRLHYGLPVVVKLLHPPSHWPLIKATVGLIRNL
                     ALCPANHAPLREQGAIPRLVQLLVRAHQDTQRRTSMGGTQQQFVEGVRMEEIVEGCTGA
                     LHILARDVHNRIVIRGLNTIPLFVQLLYSPIENIQRVAAGVLCELAQDKEAAEAIEAEG
                     ATAPLTELLHSRNEGVATYAAAVLFRMSEDKPQDYKKRLSVELTSSLFRTEPMAWNETA
                     DLGLDIGAQGEPLGYRQDDPSYRSFHSGGYGQDALGMDPMMEHEMGGHHPGADYPVDGL
                     PDLGHAQDLMDGLPPGDSNQLAWFDTDL*"
                     /label="CTNNB1"
     polyA_signal    3708..3932
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     protein_bind    4002..4035
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
                     /label="loxP"
     primer_bind     complement(4493..4509)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     promoter        4983..5087
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             5088..5948
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     rep_origin      6119..6707
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    6995..7016
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        7031..7061
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7069..7085
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     primer_bind     7093..7109
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7607..7640
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
                     /label="loxP"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtactaata
     1201 cgactcacta tagggagacc caagctggct aggtaagctt ggtaccgagc tcggatccac
     1261 tagtccagtg tggtggaatt ctgcagatat caacaagttt gtacaaaaaa gcaggctccg
     1321 cggccgcccc cttcaccatg gctactcaag ctgatttgat ggagttggac atggccatgg
     1381 aaccagacag aaaagcggct gttagtcact ggcagcaaca gtcttacctg gactctggaa
     1441 tccattctgg tgccactacc acagctcctt ctctgagtgg taaaggcaat cctgaggaag
     1501 aggatgtgga tacctcccaa gtcctgtatg agtgggaaca gggattttct cagtccttca
     1561 ctcaagaaca agtagctgat attgatggac agtatgcaat gactcgagct cagagggtac
     1621 gagctgctat gttccctgag acattagatg agggcatgca gatcccatct acacagtttg
     1681 atgctgctca tcccactaat gtccagcgtt tggctgaacc atcacagatg ctgaaacatg
     1741 cagttgtaaa cttgattaac tatcaagatg atgcagaact tgccacacgt gcaatccctg
     1801 aactgacaaa actgctaaat gacgaggacc aggtggtggt taataaggct gcagttatgg
     1861 tccatcagct ttctaaaaag gaagcttcca gacacgctat catgcgttct cctcagatgg
     1921 tgtctgctat tgtacgtacc atgcagaata caaatgatgt agaaacagct cgttgtaccg
     1981 ctgggacctt gcataacctt tcccatcatc gtgagggctt actggccatc tttaagtctg
     2041 gaggcattcc tgccctggtg aaaatgcttg gttcaccagt ggattctgtg ttgttttatg
     2101 ccattacaac tctccacaac cttttattac atcaagaagg agctaaaatg gcagtgcgtt
     2161 tagctggtgg gctgcagaaa atggttgcct tgctcaacaa aacaaatgtt aaattcttgg
     2221 ctattacgac agactgcctt caaattttag cttatggcaa ccaagaaagc aagctcatca
     2281 tactggctag tggtggaccc caagctttag taaatataat gaggacctat acttacgaaa
     2341 aactactgtg gaccacaagc agagtgctga aggtgctatc tgtctgctct agtaataagc
     2401 cggctattgt agaagctggt ggaatgcaag ctttaggact tcacctgaca gatccaagtc
     2461 aacgtcttgt tcagaactgt ctttggactc tcaggaatct ttcagatgct gcaactaaac
     2521 aggaagggat ggaaggtctc cttgggactc ttgttcagct tctgggttca gatgatataa
     2581 atgtggtcac ctgtgcagct ggaattcttt ctaacctcac ttgcaataat tataagaaca
     2641 agatgatggt ctgccaagtg ggtggtatag aggctcttgt gcgtactgtc cttcgggctg
     2701 gtgacaggga agacatcact gagcctgcca tctgtgctct tcgtcatctg accagccgac
     2761 accaagaagc agagatggcc cagaatgcag ttcgccttca ctatggacta ccagttgtgg
     2821 ttaagctctt acacccacca tcccactggc ctctgataaa ggctactgtt ggattgattc
     2881 gaaatcttgc cctttgtccc gcaaatcatg cacctttgcg tgagcagggt gccattccac
     2941 gactagttca gttgcttgtt cgtgcacatc aggataccca gcgccgtacg tccatgggtg
     3001 ggacacagca gcaatttgtg gagggggtcc gcatggaaga aatagttgaa ggttgtaccg
     3061 gagcccttca catcctagct cgggatgttc acaaccgaat tgttatcaga ggactaaata
     3121 ccattccatt gtttgtgcag ctgctttatt ctcccattga aaacatccaa agagtagctg
     3181 caggggtcct ctgtgaactt gctcaggaca aggaagctgc agaagctatt gaagctgagg
     3241 gagccacagc tcctctgaca gagttacttc actctaggaa tgaaggtgtg gcgacatatg
     3301 cagctgctgt tttgttccga atgtctgagg acaagccaca agattacaag aaacggcttt
     3361 cagttgagct gaccagctct ctcttcagaa cagagccaat ggcttggaat gagactgctg
     3421 atcttggact tgatattggt gcccagggag aaccccttgg atatcgccag gatgatccta
     3481 gctatcgttc ttttcactct ggtggatatg gccaggatgc cttgggtatg gaccccatga
     3541 tggaacatga gatgggtggc caccaccctg gtgctgacta tccagttgat gggctgccag
     3601 atctggggca tgcccaggac ctcatggatg ggctgcctcc aggtgacagc aatcagctgg
     3661 cctggtttga tactgacctg taatttaaac ccgctgatca gcctcgactg tgccttctag
     3721 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     3781 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     3841 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     3901 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     3961 ctctaggggg tatccccacg cgccctgtag cggcgcatta aataacttcg tataatgtat
     4021 gctatacgaa gttatatcgt atgcatgctt tgcatacttc tgcctgctgg ggagcctggg
     4081 gactttccac accctaactg acacacattc cacagaattc cttgaaatac atccacaggt
     4141 acacctccaa ttgactcaaa tgatgtcaat tagtctatca gaagcttcta aagccatgac
     4201 atcattttct ggaattttcc aagctgttta aaggcacagt caacttagtg tatgtaaact
     4261 tctgacccac tggaattgtg atacagtgaa ttataagtga aataatctgt ctgtaaacaa
     4321 ttgttggaaa aatgacttgt gtcatgcaca aagtagatgt cctaactgac ttgccaaaac
     4381 tattgtttgt taacaagaaa tttgtggagt agttgaaaaa cgagttttaa tgactccaac
     4441 ttaagtgtat gtaaacttcc gacttcaact gtagggtacc gagctcgaat tcactggccg
     4501 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     4561 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     4621 aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc
     4681 tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat
     4741 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     4801 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     4861 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     4921 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     4981 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     5041 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     5101 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     5161 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     5221 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     5281 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     5341 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     5401 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5461 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     5521 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5581 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5641 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5701 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5761 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5821 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5881 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5941 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     6001 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     6061 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6121 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6181 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     6241 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     6301 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6361 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6421 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6481 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6541 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6601 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6661 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6721 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6781 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     6841 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     6901 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     6961 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     7021 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     7081 aacaatttca cacaggaaac agctatgacc atgattacgc caagcttgca tgcctgcagg
     7141 tcgactctag acctatacag ttgaagtcgg aagtttacat acacttaagt tggagtcatt
     7201 aaaactcgtt tttcaactac tccacaaatt tcttgttaac aaacaatagt tttggcaagt
     7261 cagttaggac atctactttg tgcatgacac aagtcatttt tccaacaatt gtttacagac
     7321 agattatttc acttataatt cactgtatca caattccagt gggtcagaag tttacataca
     7381 ctaagttgac tgtgccttta aacagcttgg aaaattccag aaaatgatgt catggcttta
     7441 gaagcttctg atagactaat tgacatcatt tgagtcaatt ggaggtgtac ctgtggatgt
     7501 atttcaaggg gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     7561 aatgcagcgg ccgctagctc gagatcttaa ttaaactagt atcgatataa cttcgtataa
     7621 tgtatgctat acgaagttat atgtacgggc cagatatacg cgttgacatt gattattgac
     7681 taggcttttg caaaaagctt tgcaaagatg gataaagttt taaacagaga ggaatctttg
     7741 cagctaatgg accttctagg tcttgaaagg agtgggaatt
//
