LOCUS       V034679                 7066 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034679
VERSION     V034679
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"
     CDS             964..2556
                     /label="ZGPAT(NM_032527)"
                     /note="ZGPAT(NM_032527)"
                     /gene="ZGPAT"
     CDS             2563..2628
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2666..2890
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2936..3364
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3378..3707
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3774..4568
                     /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    4742..4863
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4912..4928)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4936..4952
                     /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(4960..4990)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5005..5026
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5314..5899)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6070..6930)
                     /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(6931..7035)
                     /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 aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatggacg aggagagcct ggagtcggcc ttgcagacct accgtgcgca gctgcagcag
     1021 gtggagctgg ccttgggcgc cggcctggat tcgtctgagc aggctgacct gcgccagctg
     1081 cagggggacc tgaaggagct catcgagctc accgaggcca gcctggtgtc tgtcaggaag
     1141 agcagcttgt tggccgcgct ggacgaagag cgcccgggcc gccaggaaga tgctgagtac
     1201 caggctttcc gggaggccat cactgaggcg gtggaggcac cagcagcggc ccgtgggtcc
     1261 ggatcagaga ccgttcctaa agcagaggcg gggccagaat ctgcggcagg tgggcaggag
     1321 gaggaagagg gagaggacga ggaagagctg agtgggacaa aggtgagcgc gccctactac
     1381 agctcctggg gcactctgga gtatcacaac gccatggtgg tgggaacgga agaggcggag
     1441 gatggctcgg cgggtgtccg tgtgctttac ctgtacccca ctcacaagtc tctgaagccg
     1501 tgcccgttct tcctggaggg aaagtgccgc tttaaggaga actgcaggtt ctcccatggg
     1561 caggtggtct ctctggatga gctgcgcccc ttccaggacc cagacctgag ctccctgcag
     1621 gccggctctg cgtgtctggc caagcaccag gatggcctct ggcacgcagc acgcatcacc
     1681 gatgtggaca acggctacta cacagtcaag tttgactcgc tgctgctgag ggaggccgtg
     1741 gtggaggggg acggcatcct gcccccactg cgcacagagg ccacagagtc cgactcagac
     1801 agcgacggta cgggtgactc cagctatgcc agagtggtgg ggtcggacgc cgtggactct
     1861 gcacagtcct ctgccctctg tccgtctctt gcagtggtgg ggtcagatgc tgtggactct
     1921 gggacctgca gctctgcctt tgctggctgg gaggtgcaca cgcgaggtat aggctccaga
     1981 ctcctcacca agatgggcta tgagtttggc aagggtttgg gccgacacgc ggaaggccgg
     2041 gtggagccca tccatgctgt ggtgttgcct cgagggaagt cgctggacca gtgtgtggag
     2101 accctgcaga agcagaccag ggttggcaag gctggcacca acaagccccc caggtgccgg
     2161 ggaagagggg ccaggcctgg gggccgccca gctcctcgga atgtgtttga cttcctcaat
     2221 gaaaagctgc aaggtcaggc tcctggggcc ctagaagccg gggcggcccc agcggggagg
     2281 aggagcaagg acatgtacca tgccagcaag agtgccaagc gggccctgag cctgcggctc
     2341 ttccagactg aggagaagat cgagcgaacc cagcgggaca tcaggagcat ccaggaggct
     2401 ctcgcccgca acgctggccg gcatagcgtg gcgtcagccc agctgcagga gaagctggca
     2461 ggagcccagc gccagctggg gcagctccgg gctcaggaag ccggcctgca gcaggagcag
     2521 aggaaggcag acacccacaa gaagatgact gagttcctcg aggactacaa agaccatgac
     2581 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc
     2641 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     2701 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     2761 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     2821 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     2881 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     2941 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3001 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3061 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3121 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3181 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3241 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3301 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3361 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3421 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3481 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3541 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3601 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     3661 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     3721 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     3781 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3841 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     3901 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     3961 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4021 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4081 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4141 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4201 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4261 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     4321 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4381 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4441 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4501 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4561 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     4621 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4681 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4741 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4801 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4861 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     4921 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4981 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5041 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5101 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5161 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5221 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5281 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5341 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5401 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5461 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5521 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5581 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5641 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5701 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5761 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5821 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg
     5881 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5941 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6001 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6061 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6121 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6181 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6241 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6301 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6361 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6421 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6481 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6541 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6601 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6661 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6721 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6781 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6841 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6901 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6961 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7021 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
