LOCUS       V021083                 6733 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021083
VERSION     V021083
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             932..2170
                     /label="ZDHHC6(NM_022494)"
                     /note="ZDHHC6(NM_022494)"
                     /gene="ZDHHC6"
     CDS             2231..2272
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /label="V5 tag"
     CDS             2282..2299
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    2328..2552
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2598..3026
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3040..3369
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3436..4230
                     /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    4406..4527
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4576..4592)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4600..4616
                     /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(4624..4654)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4669..4690
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4978..5566)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5737..6597)
                     /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(6598..6702)
                     /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 gctggctagt
      901 taagcttggt accgagctcg gatccgccac catgggtaca ttctgttcgg ttatcaagtt
      961 tgaaaatcta caagaattaa agagactgtg tcactggggt cccatcatag cccttggtgt
     1021 tatagcaata tgttctacca tggccatgat tgactctgtg ttgtggtatt ggcccttaca
     1081 tacaactgga ggaagtgtga atttcatcat gttgataaat tggactgtca tgattcttta
     1141 taattacttc aatgccatgt ttgtcggtcc gggctttgtc cctctggggt ggaaaccgga
     1201 aatttctcag gataccatgt atctccagta ttgtaaagtc tgccaagcat acaaggcacc
     1261 acgttcacat cactgcagaa agtgtaacag atgtgtgatg aagatggacc atcactgtcc
     1321 ttggatcaac aactgttgtg gttaccaaaa tcatgcttcg ttcacactgt ttctcctttt
     1381 agcaccactg ggttgtatcc atgctgcttt catttttgtg atgactatgt acacacagct
     1441 ttatcatcgg ctctcctttg ggtggaacac agtgaagatc gacatgagtg cagcccggag
     1501 agatcctctt ccaattgttc catttggatt agctgcattt gctaccacct tgtttgcctt
     1561 gggattagct ttaggaacaa ccatagctgt tgggatgttg ttttttatcc agatgaaaat
     1621 aattctcaga aacaaaactt ctattgagtc atggattgaa gagaaggcta aagatcgaat
     1681 tcagtattat caactagatg aagtctttgt ttttccatat gatatgggaa gtagatggag
     1741 gaactttaaa caggtattta cgtggtcagg ggtccctgaa ggagatggac ttgagtggcc
     1801 agtaagagaa ggctgtcacc aatacagctt aacaatagaa cagttgaaac aaaaagcaga
     1861 taagagagtc agaagtgttc gctataaagt aatagaagat tatagtggtg cctgctgccc
     1921 tctgaataaa ggaatcaaaa ccttcttcac aagtccctgc accgaagagc ctcgaataca
     1981 gctgcaaaaa ggggaattca ttttagccac aagaggttta cgatactggt tatatggaga
     2041 caaaattctt gatgattcct ttatagaagg tgtttcaaga ataaggggtt ggttccctag
     2101 aaaatgtgtg gaaaagtgtc cctgtgatgc tgaaacagat caagccccag agggggagaa
     2161 gaaaaataga aattctgcag atatccagca cagtggcggc cgctcgagtc tagagggccc
     2221 gcggttcgaa ggtaagccta tccctaaccc tctcctcggt ctcgattcta cgcgtaccgg
     2281 tcatcatcac catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctag
     2341 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     2401 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     2461 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     2521 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     2581 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2641 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2701 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     2761 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     2821 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     2881 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     2941 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     3001 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     3061 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3121 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3181 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3241 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3301 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3361 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     3421 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3481 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3541 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3601 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3661 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3721 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3781 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3841 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     3901 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     3961 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4021 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4081 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4141 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4201 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga
     4261 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     4321 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4381 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     4441 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4501 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     4561 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4621 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4681 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4741 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4801 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4861 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4921 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4981 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5041 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5101 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5161 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5221 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5281 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5341 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5401 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5461 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5521 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5581 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5641 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5701 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5761 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5821 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5881 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5941 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6001 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6061 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6121 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6181 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6241 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6301 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6361 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6421 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6481 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6541 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6601 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6661 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6721 gccacctgac gtc
//
