LOCUS       62056_421        7856 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7856)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7856
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             903..926
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     CDS             957..3299
                     /codon_start=1
                     /label=CTNNB1
                     /note="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"
                     /translation="MATQADLMELDMAMEPDRKAAVSHWQQQSYLDAGIHAGATATAPA
                     LSGKGNPEEEDVDTSQVLYEWEQGFSQSFTQEQVADIDGQYAMTRAQRVRAAMFPETLD
                     EGMQIPSTQFDAAHPTNVQRLAEPSQMLKHAVVNLINYQDDAELATRAIPELTKLLNDE
                     DQVVVNKAAVMVHQLSKKEASRHAIMRSPQMVSAIVRTMQNTNDVETARCTAGTLHNLS
                     HHREGLLAIFKSGGIPALVKMLGSPVDSVLFYAITTLHNLLLHQEGAKMAVRLAGGLQK
                     MVALLNKTNVKFLAITTDCLQILAYGNQESKLIILASGGPQALVNIMRTYTYEKLLWTT
                     SRVLKVLSVCSSNKPAIVEAGGMQALGLHLTDPSQRLVQNCLWTLRNLSDAATKQEGME
                     GLLGTLVQLLGSDDINVVTCAAGILSNLTCNNYKNKMMVCQVGGIEALVRTVLRAGDRE
                     DITEPAICALRHLTSRHQEAEMAQNAVRLHYGLPVVVKLLHPPSHWPLIKATVGLIRNL
                     ALCPANHAPLREQGAIPRLVQLLVRAHQDTQRRTSMGGTQQQFVEGVRMEEIVEGCTGA
                     LHILARDVHNRIVIRGLNTIPLFVQLLYSPIENIQRVAAGVLCELAQDKEAAEAIEAEG
                     ATAPLTELLHSRNEGVATYAAAVLFRMSEDKPQDYKKRLSVELTSSLFRTEPMAWNETA
                     DLGLDIGAQGEPLGYRQDDPSYRSFHSGGYGQDALGMDPMMEHEMGGHHPGADYPVDGL
                     PDLGHAQDLMDGLPPGDSNQLAWFDTDL"
     promoter        complement(3408..3426)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    3452..3676
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3722..4150
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4164..4494
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4561..5352
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5529..5662
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5699..5715)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5723..5739)
                     /label=lac operator
                     /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(5747..5777)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5792..5813)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6101..6689)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6863..7720)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7721..7825)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctgccacca
      901 tggactacaa ggacgacgat gacaagggac ctaagcttgg taccgagctc ggatccatgg
      961 ctactcaagc tgatttgatg gagttggaca tggccatgga accagacaga aaagcggctg
     1021 ttagtcactg gcagcaacag tcttacctgg acgctggaat ccatgctggt gccactgcca
     1081 cagctcctgc tctgagtggt aaaggcaatc ctgaggaaga ggatgtggat acctcccaag
     1141 tcctgtatga gtgggaacag ggattttctc agtccttcac tcaagaacaa gtagctgata
     1201 ttgatggaca gtatgcaatg actcgagctc agagggtacg agctgctatg ttccctgaga
     1261 cattagatga gggcatgcag atcccatcta cacagtttga tgctgctcat cccactaatg
     1321 tccagcgttt ggctgaacca tcacagatgc tgaaacatgc agttgtaaac ttgattaact
     1381 atcaagatga tgcagaactt gccacacgtg caatccctga actgacaaaa ctgctaaatg
     1441 acgaggacca ggtggtggtt aataaggctg cagttatggt ccatcagctt tctaaaaagg
     1501 aagcttccag acacgctatc atgcgttctc ctcagatggt gtctgctatt gtacgtacca
     1561 tgcagaatac aaatgatgta gaaacagctc gttgtaccgc tgggaccttg cataaccttt
     1621 cccatcatcg tgagggctta ctggccatct ttaagtctgg aggcattcct gccctggtga
     1681 aaatgcttgg ttcaccagtg gattctgtgt tgttttatgc cattacaact ctccacaacc
     1741 ttttattaca tcaagaagga gctaaaatgg cagtgcgttt agctggtggg ctgcagaaaa
     1801 tggttgcctt gctcaacaaa acaaatgtta aattcttggc tattacgaca gactgccttc
     1861 aaattttagc ttatggcaac caagaaagca agctcatcat actggctagt ggtggacccc
     1921 aagctttagt aaatataatg aggacctata cttacgaaaa actactgtgg accacaagca
     1981 gagtgctgaa ggtgctatct gtctgctcta gtaataagcc ggctattgta gaagctggtg
     2041 gaatgcaagc tttaggactt cacctgacag atccaagtca acgtcttgtt cagaactgtc
     2101 tttggactct caggaatctt tcagatgctg caactaaaca ggaagggatg gaaggtctcc
     2161 ttgggactct tgttcagctt ctgggttcag atgatataaa tgtggtcacc tgtgcagctg
     2221 gaattctttc taacctcact tgcaataatt ataagaacaa gatgatggtc tgccaagtgg
     2281 gtggtataga ggctcttgtg cgtactgtcc ttcgggctgg tgacagggaa gacatcactg
     2341 agcctgccat ctgtgctctt cgtcatctga ccagccgaca ccaagaagca gagatggccc
     2401 agaatgcagt tcgccttcac tatggactac cagttgtggt taagctctta cacccaccat
     2461 cccactggcc tctgataaag gctactgttg gattgattcg aaatcttgcc ctttgtcccg
     2521 caaatcatgc acctttgcgt gagcagggtg ccattccacg actagttcag ttgcttgttc
     2581 gtgcacatca ggatacccag cgccgtacgt ccatgggtgg gacacagcag caatttgtgg
     2641 agggggtccg catggaagaa atagttgaag gttgtaccgg agcccttcac atcctagctc
     2701 gggatgttca caaccgaatt gttatcagag gactaaatac cattccattg tttgtgcagc
     2761 tgctttattc tcccattgaa aacatccaaa gagtagctgc aggggtcctc tgtgaacttg
     2821 ctcaggacaa ggaagctgca gaagctattg aagctgaggg agccacagct cctctgacag
     2881 agttacttca ctctaggaat gaaggtgtgg cgacatatgc agctgctgtt ttgttccgaa
     2941 tgtctgagga caagccacaa gattacaaga aacggctttc agttgagctg accagctctc
     3001 tcttcagaac agagccaatg gcttggaatg agactgctga tcttggactt gatattggtg
     3061 cccagggaga accccttgga tatcgccagg atgatcctag ctatcgttct tttcactctg
     3121 gtggatatgg ccaggatgcc ttgggtatgg accccatgat ggaacatgag atgggtggcc
     3181 accaccctgg tgctgactat ccagttgatg ggctgccaga tctggggcat gcccaggacc
     3241 tcatggatgg gctgcctcca ggtgacagca atcagctggc ctggtttgat actgacctgt
     3301 aaatcatcct ttaggaggat ccactagtaa cggccgccag tgtgctggaa ttctgcagat
     3361 atccatcaca ctggcggccg ctcgagcatg catctagagg gccctattct atagtgtcac
     3421 ctaaatgcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3481 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3541 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3601 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3661 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     3721 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3781 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3841 acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt
     3901 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3961 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4021 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4081 taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4141 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4201 caggcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4261 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4321 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4381 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     4441 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     4501 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     4561 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4621 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4681 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4741 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4801 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4861 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4921 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4981 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     5041 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     5101 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     5161 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     5221 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5281 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5341 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     5401 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5461 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5521 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5581 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5641 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     5701 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5761 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5821 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5881 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5941 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6001 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6061 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6121 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6181 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6241 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct
     6301 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6361 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6421 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6481 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6541 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6601 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6661 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6721 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6781 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6841 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6901 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6961 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7021 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7081 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7141 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7201 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7261 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7321 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7381 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7441 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7501 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7561 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7621 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7681 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7741 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7801 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
