LOCUS       V018667                 8150 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018667
VERSION     V018667
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             1003..3696
                     /label="DPP8(NM_197960)"
                     /note="DPP8(NM_197960)"
                     /gene="DPP8"
     polyA_signal    3750..3974
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4020..4448
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4462..4791
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4858..5652
                     /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    5826..5947
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5996..6012)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6020..6036
                     /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(6044..6074)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6089..6110
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6398..6983)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7154..8014)
                     /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(8015..8119)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 gctggctagc
      901 gtttaaactt aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt ggtaccgcca ccatgtggaa gagatctgag
     1021 cagatgaaaa taaaatcagg aaaatgcaac atggcagcag caatggaaac agaacagctg
     1081 ggtgttgaga tatttgaaac tgcggactgt gaggagaata ttgaatcaca ggatcggcct
     1141 aaattggagc ctttttatgt tgagcggtat tcctggagtc agcttaaaaa gctgcttgcc
     1201 gataccagaa aatatcatgg ctacatgatg gctaaggcac cacatgattt catgtttgtg
     1261 aagaggaatg atccagatgg acctcattca gacagaatct attaccttgc catgtctggt
     1321 gagaacagag aaaatacact gttttattct gaaattccca aaactatcaa tagagcagca
     1381 gtcttaatgc tctcttggaa gcctcttttg gatctttttc aggcaacact ggactatgga
     1441 atgtattctc gagaagaaga actattaaga gaaagaaaac gcattggaac agtcggaatt
     1501 gcttcttacg attatcacca aggaagtgga acatttctgt ttcaagccgg tagtggaatt
     1561 tatcacgtaa aagatggagg gccacaagga tttacgcaac aacctttaag gcccaatcta
     1621 gtggaaacta gttgtcccaa catacggatg gatccaaaat tatgccctgc tgatccagac
     1681 tggattgctt ttatacatag caacgatatt tggatatcta acatcgtaac cagagaagaa
     1741 aggagactca cttatgtgca caatgagcta gccaacatgg aagaagatgc cagatcagct
     1801 ggagtcgcta cctttgttct ccaagaagaa tttgatagat attctggcta ttggtggtgt
     1861 ccaaaagctg aaacaactcc cagtggtggt aaaattctta gaattctata tgaagaaaat
     1921 gatgaatctg aggtggaaat tattcatgtt acatccccta tgttggaaac aaggagggca
     1981 gattcattcc gttatcctaa aacaggtaca gcaaatccta aagtcacttt taagatgtca
     2041 gaaataatga ttgatgctga aggaaggatc atagatgtca tagataagga actaattcaa
     2101 ccttttgaga ttctatttga aggagttgaa tatattgcca gagctggatg gactcctgag
     2161 ggaaaatatg cttggtccat cctactagat cgctcccaga ctcgcctaca gatagtgttg
     2221 atctcacctg aattatttat cccagtagaa gatgatgtta tggaaaggca gagactcatt
     2281 gagtcagtgc ctgattctgt gacgccacta attatctatg aagaaacaac agacatctgg
     2341 ataaatatcc atgacatctt tcatgttttt ccccaaagtc acgaagagga aattgagttt
     2401 atttttgcct ctgaatgcaa aacaggtttc cgtcatttat acaaaattac atctatttta
     2461 aaggaaagca aatataaacg atccagtggt gggctgcctg ctccaagtga tttcaagtgt
     2521 cctatcaaag aggagatagc aattaccagt ggtgaatggg aagttcttgg ccggcatgga
     2581 tctaatatcc aagttgatga agtcagaagg ctggtatatt ttgaaggcac caaagactcc
     2641 cctttagagc atcacctgta cgtagtcagt tacgtaaatc ctggagaggt gacaaggctg
     2701 actgaccgtg gctactcaca ttcttgctgc atcagtcagc actgtgactt ctttataagt
     2761 aagtatagta accagaagaa tccacactgt gtgtcccttt acaagctatc aagtcctgaa
     2821 gatgacccaa cttgcaaaac aaaggaattt tgggccacca ttttggattc agcaggtcct
     2881 cttcctgact atactcctcc agaaattttc tcttttgaaa gtactactgg atttacattg
     2941 tatgggatgc tctacaagcc tcatgatcta cagcctggaa agaaatatcc tactgtgctg
     3001 ttcatatatg gtggtcctca ggtgcagttg gtgaataatc ggtttaaagg agtcaagtat
     3061 ttccgcttga ataccctagc ctctctaggt tatgtggttg tagtgataga caacagggga
     3121 tcctgtcacc gagggcttaa atttgaaggc gcctttaaat ataaaatggg tcaaatagaa
     3181 attgacgatc aggtggaagg actccaatat ctagcttctc gatatgattt cattgactta
     3241 gatcgtgtgg gcatccacgg ctggtcctat ggaggatacc tctccctgat ggcattaatg
     3301 cagaggtcag atatcttcag ggttgctatt gctggggccc cagtcactct gtggatcttc
     3361 tatgatacag gatacacgga acgttatatg ggtcaccctg accagaatga acagggctat
     3421 tacttaggat ctgtggccat gcaagcagaa aagttcccct ctgaaccaaa tcgtttactg
     3481 ctcttacatg gtttcctgga tgagaatgtc cattttgcac ataccagtat attactgagt
     3541 tttttagtga gggctggaaa gccatatgat ttacagatct atcctcagga gagacacagc
     3601 ataagagttc ctgaatcggg agaacattat gaactgcatc ttttgcacta ccttcaagaa
     3661 aaccttggat cacgtattgc tgctctaaaa gtgatataag cggccgctcg agtctagagg
     3721 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     3781 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     3841 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     3901 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     3961 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     4021 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4081 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4141 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4201 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4261 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4321 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4381 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4441 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4501 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4561 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4621 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4681 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     4741 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4801 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4861 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4921 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4981 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     5041 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     5101 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     5161 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     5221 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     5281 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     5341 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     5401 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     5461 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5521 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5581 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5641 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     5701 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     5761 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     5821 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5881 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5941 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     6001 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     6061 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     6121 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     6181 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6241 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6301 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6361 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6421 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6481 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6541 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6601 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6661 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6721 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6781 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6841 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6901 cttgatccgg caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta
     6961 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     7021 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     7081 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     7141 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     7201 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     7261 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     7321 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     7381 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     7441 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     7501 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7561 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7621 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7681 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7741 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     7801 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7861 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7921 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7981 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     8041 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     8101 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
