LOCUS       V028463                 7116 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028463
VERSION     V028463
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     primer_bind     141..157
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     enhancer        318..697
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        698..901
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             931..996
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             1000..1821
                     /label="CCS(NM_005125)"
                     /note="CCS(NM_005125)"
                     /gene="CCS"
     polyA_signal    1875..2497
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     promoter        2539..2836
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             2890..3684
                     /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    4336..4470
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     promoter        complement(4476..4494)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(4508..4524)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4532..4548
                     /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(4556..4586)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4601..4622
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4910..5498)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5669..6529)
                     /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(6530..6634)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      6661..7089
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa
      961 gatcatgaca tcgattacaa ggatgacgat gacaagctta tggcttcgga ttcggggaac
     1021 caggggaccc tctgcacgtt ggagttcgcg gtgcagatga cctgtcagag ctgtgtggac
     1081 gcggtgcgca aatccctgca aggggtggca ggtgtccagg atgtggaggt gcacttggag
     1141 gaccagatgg tcttggtaca caccactcta cccagccagg aggtgcaggc tctcctggaa
     1201 ggcacggggc ggcaggcggt actcaagggc atgggcagcg gccagttgca gaatctgggg
     1261 gcagcagtgg ccatcctggg ggggcctggc accgtgcagg gggtggtgcg cttcctacag
     1321 ctgacccctg agcgctgcct catcgaggga actattgacg gcctggagcc tgggctgcat
     1381 ggactccacg tccatcagta cggggacctt acaaacaact gcaacagctg tgggaatcac
     1441 tttaaccctg atggagcatc tcatgggggc ccccaggact ctgaccggca ccgcggagac
     1501 ctgggcaatg tccgtgctga tgctgacggc cgcgccatct tcagaatgga ggatgagcag
     1561 ctgaaggtgt gggatgtgat tggccgcagc ctgattattg atgagggaga agatgacctg
     1621 ggccggggag gccatccctt atccaagatc acagggaact ccggggagag gttggcctgt
     1681 ggcatcattg cacgctccgc tggccttttc cagaacccca agcagatctg ctcttgcgat
     1741 ggcctcacca tctgggagga gcgaggccgg cccatcgctg gcaagggccg aaaggagtca
     1801 gcgcagcccc ctgcccacct ttgagaattc atcgatagat ctgatatcgg taccagtcga
     1861 ctctagagga tcccgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg
     1921 aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt
     1981 ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc
     2041 aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag
     2101 tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc
     2161 agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt
     2221 tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg
     2281 tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct
     2341 tccctgtcct tctgatttta aaataactat accagcagga ggacgtccag acacagcata
     2401 ggctacctgg ccatgcccaa ccggtgggac atttgagttg cttgcttggc actgtcctct
     2461 catgcgttgg gtccactcag tagatgcctg ttgaattggg tacgcggcca gcttggctgt
     2521 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2581 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     2641 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     2701 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     2761 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     2821 gaggaggctt ttttggagga attgatcagc ttgggatctg atcaagagac aggatgagga
     2881 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     2941 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3001 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3061 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3121 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3181 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     3241 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     3301 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     3361 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     3421 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     3481 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     3541 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     3601 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     3661 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     3721 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     3781 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     3841 agttcttcgc ccaccccggg ctcgatcccc tcgcgagttg gttcagctgc tgcctgaggc
     3901 tggacgacct cgcggagttc taccggcagt gcaaatccgt cggcatccag gaaaccagca
     3961 gcggctatcc gcgcatccat gcccccgaac tgcaggagtg gggaggcacg atggccgctt
     4021 tggtcgaccc ggacgggacg ctcctgcgcc tgatacagaa cgaattgctt gcaggcatct
     4081 catgagtgtg tcttcccgtt ttccgcctga ggtcactgcg tggatggagc gctggcgcct
     4141 gctgcgcgac ggcgagctgc tcaccaccca ctcgccaagc tggaaccgta aaaaggccgc
     4201 gttgctggcg tttttccata ggctccgccg atcataatca gccataccac atttgtagag
     4261 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     4321 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     4381 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     4441 ctcatcaatg tatcttatca tgtctggatc aattccctat agtgagtcgt attaaattcg
     4501 taatcatgtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4561 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4621 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4681 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4741 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4801 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4861 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4921 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4981 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5041 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5101 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5161 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5221 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5281 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5341 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5401 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5461 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5521 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5581 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5641 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5701 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5761 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5821 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5881 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5941 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6001 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6061 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6121 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6181 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6241 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6301 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6361 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6421 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6481 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6541 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6601 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6661 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     6721 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     6781 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     6841 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     6901 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     6961 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     7021 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     7081 acgcgaattt taacaaaata ttaacgctta caattt
//
