LOCUS       V033867                 8693 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033867
VERSION     V033867
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             884..1591
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             1628..4213
                     /label="AXIN1(NM_003502)"
                     /note="AXIN1(NM_003502)"
                     /gene="AXIN1"
     polyA_signal    4260..4484
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4530..4958
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4972..5301
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5368..6162
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    6336..6457
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6506..6522)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6530..6546
                     /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"
     promoter        complement(6554..6584)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6599..6620
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6908..7493)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7664..8524)
                     /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"
     promoter        complement(8525..8629)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgct accggtcgcc accatggtga gcaagggcga
      901 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
      961 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     1021 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
     1081 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga
     1141 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     1201 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
     1261 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
     1321 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
     1381 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
     1441 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
     1501 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
     1561 ctccaccggc ggcatggacg agctgtacaa gggcggtgga ggaagcggcg gaggtggatc
     1621 tctcgagatg aatatccaag agcagggttt ccccttggac ctcggagcaa gtttcaccga
     1681 agatgctccc cgacccccag tgcctggtga ggagggagaa ctggtgtcca cagacccgag
     1741 gcccgccagc tacagtttct gctccgggaa aggtgttggc attaaaggtg agacttcgac
     1801 ggccactccg aggcgctcgg atctggacct ggggtatgag cctgagggca gtgcctcccc
     1861 caccccacca tacttgaagt gggctgagtc actgcattcc ctgctggatg accaagatgg
     1921 gataagcctg ttcaggactt tcctgaagca ggagggctgt gccgacttgc tggacttctg
     1981 gtttgcctgc actggcttca ggaagctgga gccctgtgac tcgaacgagg agaagaggct
     2041 gaagctggcg agagccatct accgaaagta cattcttgat aacaatggca tcgtgtcccg
     2101 gcagaccaag ccagccacca agagcttcat aaagggctgc atcatgaagc agctgatcga
     2161 tcctgccatg tttgaccagg cccagaccga aatccaggcc actatggagg aaaacaccta
     2221 tccctccttc cttaagtctg atatttattt ggaatatacg aggacaggct cggagagccc
     2281 caaagtctgt agtgaccaga gctctgggtc agggacaggg aagggcatat ctggatacct
     2341 gccgacctta aatgaagatg aggaatggaa gtgtgaccag gacatggatg aggacgatgg
     2401 cagagacgct gctccccccg gaagactccc tcagaagctg ctcctggaga cagctgcccc
     2461 gagggtctcc tccagtagac ggtacagcga aggcagagag ttcaggtatg gatcctggcg
     2521 ggagccagtc aacccctatt atgtcaatgc cggctatgcc ctggccccag ccaccagtgc
     2581 caacgacagc gagcagcaga gcctgtccag cgatgcagac accctgtccc tcacggacag
     2641 cagcgtggat gggatccccc catacaggat ccgtaagcag caccgcaggg agatgcagga
     2701 gagcgtgcag gtcaatgggc gggtgcccct acctcacatt ccccgcacgt accgggtgcc
     2761 gaaggaggtc cgcgtggagc ctcagaagtt cgcggaggag ctcatccacc gcctggaggc
     2821 tgtgcagcgc acgcgggagg ccgaggagaa gctggaggag cggctgaagc gcgtgcgcat
     2881 ggaggaggaa ggtgaggacg gcgatccatc gtcagggccc ccagggccgt gtcacaagct
     2941 gcctcccgcc cccgcttggc accacttccc gccccgctgt gtggacatgg gctgtgccgg
     3001 gctccgggat gcacacgagg agaaccctga gagcatcctg gacgagcacg tacagcgtgt
     3061 gctgaggaca cctggccgcc agtcgcctgg gcctggccat cgctccccgg acagtgggca
     3121 cgtggccaag atgccagtgg cactgggggg tgccgcctcg gggcacggga agcacgtacc
     3181 caagtcaggg gcgaagctgg acgcggccgg cctgcaccac caccgacacg tccaccacca
     3241 cgtccaccac agcacagccc ggcccaagga gcaggtggag gccgaggcca cccgcagggc
     3301 ccagagcagc ttcgcctggg gcctggaacc acacagccat ggggcaaggt cccgaggcta
     3361 ctcagagagt gttggcgctg cccccaacgc cagtgatggc ctcgcccaca gtgggaaggt
     3421 gggcgtggcg tgcaaaagaa atgccaagaa ggctgagtcg gggaagagcg ccagcaccga
     3481 ggtgccaggt gcctcggagg atgcggagaa gaaccagaaa atcatgcagt ggatcattga
     3541 gggggaaaag gagatcagca ggcaccgcag gaccggccac gggtcttcgg ggacgaggaa
     3601 gccacagccc catgagaact ccagaccctt gtcccttgag cacccctggg ccggccctca
     3661 gctccggacc tccgtgcagc cctcccacct cttcatccaa gaccccacca tgccacccca
     3721 cccagctccc aaccccctaa cccagctgga ggaggcgcgc cgacgtctgg aggaggaaga
     3781 aaagagagcc agccgagcac cctccaagca gaggtatgtg caggaggtta tgcggcgggg
     3841 acgcgcctgc gtcaggccag cgtgcgcgcc ggtgctgcac gtggtaccag ccgtgtcgga
     3901 catggagctc tccgagacag agacaagatc gcagaggaag gtgggcggcg ggagtgccca
     3961 gccgtgtgac agcatcgttg tggcgtacta cttctgcggg gaacccatcc cctaccgcac
     4021 cctggtgagg ggccgcgctg tcaccctggg ccagttcaag gagctgctga ccaaaaaggg
     4081 cagctacaga tactacttca agaaagtgag cgacgagttt gactgtgggg tggtgtttga
     4141 ggaggttcga gaggacgagg ccgtcctgcc cgtctttgag gagaagatca tcggcaaagt
     4201 ggagaaggtg gactgaggat cctctagagg gcccgtttaa acccgctgat cagcctcgac
     4261 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     4321 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     4381 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     4441 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     4501 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     4561 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     4621 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4681 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4741 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4801 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4861 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     4921 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     4981 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     5041 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5101 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     5161 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     5221 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     5281 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     5341 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     5401 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     5461 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     5521 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     5581 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     5641 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     5701 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     5761 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     5821 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     5881 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     5941 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     6001 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     6061 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     6121 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     6181 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     6241 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     6301 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     6361 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     6421 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     6481 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     6541 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6601 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6661 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6721 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6781 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6841 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6901 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6961 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     7021 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     7081 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     7141 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7201 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7261 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     7321 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     7381 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7441 gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     7501 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     7561 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     7621 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7681 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7741 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7801 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7861 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7921 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7981 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     8041 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     8101 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     8161 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     8221 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     8281 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     8341 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     8401 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     8461 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     8521 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     8581 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     8641 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggt
//
