LOCUS       V029499                 6550 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029499
VERSION     V029499
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             706..1206
                     /label="ZG16(NM_152338)"
                     /note="ZG16(NM_152338)"
                     /gene="ZG16"
     CDS             1213..1302
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    1326..1914
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    1980..2028
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2230..2658
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2672..3001
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3068..3862
                     /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    4038..4159
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4208..4224)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4232..4248
                     /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(4256..4286)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4301..4322
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4610..5198)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5369..6229)
                     /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(6230..6334)
                     /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 ggatccacta gtccagtgtg gtggaattcg ccaccatgtt gacagtcgct
      721 ctcctagccc ttctctgtgc ctcagcctct ggcaatgcca ttcaggccag gtcttcctcc
      781 tatagtggag agtatggagg tggtggtgga aagcgattct ctcattctgg caaccagttg
      841 gacggcccca tcaccgccct ccgggtccga gtcaacacat actacatcgt aggtcttcag
      901 gtgcgctatg gcaaggtgtg gagcgactat gtgggtggtc gcaacggaga cctggaggag
      961 atctttctgc accctgggga atcagtgatc caggtttctg ggaagtacaa gtggtacctg
     1021 aagaagctgg tatttgtgac agacaagggc cgctatctgt cttttgggaa agacagtggc
     1081 acaagtttca atgccgtccc cttgcacccc aacaccgtgc tccgcttcat cagtggccgg
     1141 tctggttctc tcatcgatgc cattggcctg cactgggatg tttaccccag tagctgcagc
     1201 agatgcctcg aggaacaaaa actaatatca gaggaagatt tggaacagaa actaatcagt
     1261 gaagaagatt tagagcaaaa gttaatttcc gaggaggact tatagtgagg gcccgatatc
     1321 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     1381 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     1441 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     1501 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     1561 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     1621 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     1681 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
     1741 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     1801 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     1861 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa
     1921 acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     1981 ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg
     2041 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     2101 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     2161 cagccaacgt cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg
     2221 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     2281 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     2341 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt
     2401 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     2461 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     2521 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     2581 ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac
     2641 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     2701 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     2761 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     2821 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     2881 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     2941 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3001 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     3061 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3121 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3181 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3241 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3301 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3361 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     3421 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     3481 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     3541 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     3601 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     3661 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     3721 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     3781 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     3841 ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga
     3901 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     3961 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4021 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4081 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4141 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     4201 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4261 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4321 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4381 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4441 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4501 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4561 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4621 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4681 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4741 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4801 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4861 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4921 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4981 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5041 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5101 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5161 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5221 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5281 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5341 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5401 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5461 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5521 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5581 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5641 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5701 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5761 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5821 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5881 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5941 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6001 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6061 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6121 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6181 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6241 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6301 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6361 acgtcgacgg atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc
     6421 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
     6481 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
     6541 tctgcttagg
//
