LOCUS       V019254                 8231 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019254
VERSION     V019254
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             683..2911
                     /label="NAGLU(NM_000263)"
                     /note="NAGLU(NM_000263)"
                     /gene="NAGLU"
     CDS             2918..2983
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3007..3595
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3661..3709
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3911..4339
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4353..4682
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4749..5543
                     /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    5719..5840
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5889..5905)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5913..5929
                     /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(5937..5967)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5982..6003
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6291..6879)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7050..7910)
                     /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(7911..8015)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatggaggc ggtggcggtg gccgcggcgg tgggggtcct
      721 tctcctggcc ggggccgggg gcgcggcagg cgacgaggcc cgggaggcgg cggccgtgcg
      781 ggcgctcgtg gcccggctgc tggggccagg ccccgcggcc gacttctccg tgtcggtgga
      841 gcgcgctctg gctgccaagc cgggcttgga cacctacagc ctgggcggcg gcggcgcggc
      901 gcgcgtgcgg gtgcgcggct ccacgggcgt ggcggccgcc gcggggctgc accgctacct
      961 gcgcgacttc tgtggctgcc acgtggcctg gtccggctct cagctgcgcc tgccgcggcc
     1021 actgccagcc gtgccggggg agctgaccga ggccacgccc aacaggtacc gctattacca
     1081 gaatgtgtgc acgcaaagct actctttcgt gtggtgggac tgggcccgct gggagcgaga
     1141 gatagactgg atggcgctga atggcatcaa cctggcactg gcctggagcg gccaggaggc
     1201 catctggcag cgggtgtacc tggccttggg cctgacccag gcagagatca atgagttctt
     1261 tactggtcct gccttcctgg cctgggggcg aatgggcaac ctgcacacct gggatggccc
     1321 cctgcccccc tcctggcaca tcaagcagct ttacctgcag caccgggtcc tggaccagat
     1381 gcgctccttc ggcatgaccc cagtgctgcc tgcattcgcg gggcatgttc ccgaggctgt
     1441 caccagggtg ttccctcagg tcaatgtcac gaagatgggc agttggggcc actttaactg
     1501 ttcctactcc tgctccttcc ttctggctcc ggaagacccc atattcccca tcatcgggag
     1561 cctcttcctg cgagagctga tcaaagagtt tggcacagac cacatctatg gggccgacac
     1621 tttcaatgag atgcagccac cttcctcaga gccctcctac cttgccgcag ccaccactgc
     1681 cgtctatgag gccatgactg cagtggatac tgaggctgtg tggctgctcc aaggctggct
     1741 cttccagcac cagccgcagt tctgggggcc cgcccagatc agggctgtgc tgggagctgt
     1801 gccccgtggc cgcctcctgg ttctggacct gtttgctgag agccagcctg tgtatacccg
     1861 cactgcctcc ttccagggcc agcccttcat ctggtgcatg ctgcacaact ttgggggaaa
     1921 ccatggtctt tttggagccc tagaggctgt gaacggaggc ccagaagctg cccgcctctt
     1981 ccccaactcc accatggtag gcacgggcat ggcccccgag ggcatcagcc agaacgaagt
     2041 ggtctattcc ctcatggctg agctgggctg gcgaaaggac ccagtgccag atttggcagc
     2101 ctgggtgacc agctttgccg cccggcggta tggggtctcc cacccggacg caggggcagc
     2161 gtggaggcta ctgctccgga gtgtgtacaa ctgctccggg gaggcctgca ggggccacaa
     2221 tcgtagcccg ctggtcaggc ggccgtccct acagatgaat accagcatct ggtacaaccg
     2281 atctgatgtg tttgaggcct ggcggctgct gctcacatct gctccctccc tggccaccag
     2341 ccccgccttc cgctacgacc tgctggacct cactcggcag gcagtgcagg agctggtcag
     2401 cttgtactat gaggaggcaa gaagcgccta cctgagcaag gagctggcct ccctgttgag
     2461 ggctggaggc gtcctggcct atgagctgct gccggcactg gacgaggtgc tggctagtga
     2521 cagccgcttc ttgctgggca gctggctaga gcaggcccga gcagcggcag tcagtgaggc
     2581 cgaggccgat ttctacgagc agaacagccg ctaccagctg accttgtggg ggccagaagg
     2641 caacatcctg gactatgcca acaagcagct ggcggggttg gtggccaact actacacccc
     2701 tcgctggcgg cttttcctgg aggcgctggt tgacagtgtg gcccagggca tccctttcca
     2761 acagcaccag tttgacaaaa atgtcttcca actggagcag gccttcgttc tcagcaagca
     2821 gaggtacccc agccagccgc gaggagacac tgtggacctg gccaagaaga tcttcctcaa
     2881 atattacccc cgctgggtgg ccggctcttg gctcgaggac tacaaagacc atgacggtga
     2941 ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag ggcccgatat
     3001 ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     3061 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     3121 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     3181 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     3241 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     3301 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     3361 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
     3421 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     3481 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     3541 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga
     3601 aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     3661 cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg
     3721 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     3781 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     3841 gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg
     3901 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3961 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     4021 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg
     4081 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     4141 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4201 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     4261 tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa
     4321 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     4381 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     4441 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4501 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4561 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4621 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4681 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     4741 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4801 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4861 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4921 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4981 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     5041 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     5101 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     5161 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     5221 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     5281 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     5341 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     5401 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     5461 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     5521 gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg
     5581 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     5641 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     5701 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5761 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5821 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     5881 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5941 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6001 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6061 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6121 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6181 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6241 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6301 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6361 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6421 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6481 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6541 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6601 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6661 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6721 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6781 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6841 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6901 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6961 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7021 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7081 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7141 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7201 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7261 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7321 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7381 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7441 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7501 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7561 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7621 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     7681 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     7741 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7801 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7861 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7921 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7981 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     8041 gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt acaatctgct
     8101 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     8161 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     8221 atctgcttag g
//
