LOCUS       V024338                 7571 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024338
VERSION     V024338
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             941..3061
                     /label="ZCCHC8(NM_017612)"
                     /note="ZCCHC8(NM_017612)"
                     /gene="ZCCHC8"
     CDS             3068..3133
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3171..3395
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3441..3869
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3883..4212
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4279..5073
                     /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    5247..5368
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5417..5433)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5441..5457
                     /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(5465..5495)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5510..5531
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5819..6404)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6575..7435)
                     /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(7436..7540)
                     /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 atccgccacc atggccgcag aggtgtattt
      961 tggcgatcta gagctcttcg agccgttcga ccacccagag gagtcgattc cgaagcccgt
     1021 tcacactcgc ttcaaggacg acgacggcga cgaggaggac gaaaatgggg tcggcgacgc
     1081 ggagctacgg gagcggcttc ggcagtgcga ggagaccatc gagcagctcc gcgccgagaa
     1141 tcaagaactt aaacgaaaat tgaacattct gactcgaccg agtggaatat tggtgaacga
     1201 tactaagtta gatggaccta tattacagat tctattcatg aacaatgcta tttcaaagca
     1261 atatcatcaa gaaatagagg aatttgtatc aaatttagta aaaagatttg aggaacagca
     1321 gaaaaatgat gtggaaaaga cttcctttaa tcttttgccc cagccatcca gtattgtgct
     1381 agaggaggac cacaaagtgg aagagtcctg tgccattaaa aacaacaagg aagctttcag
     1441 tgttgtagga agtgtcctgt attttactaa tttttgcctt gataaattgg ggcaaccgct
     1501 tctaaatgaa aaccctcagc tttccgaagg atgggaaata cccaagtacc atcaagtctt
     1561 cagccacatt gtttctctag aagggcaaga aatacaagta aaggcaaaaa ggccaaagcc
     1621 tcactgtttc aattgtggtt ctgaagaaca ccaaatgaaa gattgcccaa tgcctcggaa
     1681 tgctgctcga ataagtgaaa agagaaaaga gtatatggat gcctgtggag aagcaaacaa
     1741 tcagaatttc cagcagcgat accacgcaga agaagtagaa gaaagatttg gaagattcaa
     1801 gccaggagtt attagtgagg aacttcaaga tgcactaggt gtgacagaca agagtcttcc
     1861 accttttatc tatcggatgc gccagctagg gtacccacca gggtggctca aagaggctga
     1921 attggagaat tcggggcttg cactctatga tggaaaagat ggcactgatg gggaaacaga
     1981 agttggagaa atacaacaga ataaaagtgt cacttacgat ctctcaaaat tggtcaacta
     2041 tcctggtttt aatatatcta ctcccagagg aattccagac gaatggagga tctttggttc
     2101 cataccaatg caggcatgtc agcagaagga tgtgtttgcc aattacctta cttctaactt
     2161 ccaagcgcca ggtgtgaagt ctggcaacaa gaggtcttca tctcactcta gcccaggtag
     2221 tccaaagaag cagaagaatg aaagcaactc agcgggatct cccgccgaca tggagctcga
     2281 ttcagatatg gaggtaccac atggttctca gagcagcgaa agttttcagt ttcaaccacc
     2341 attacctcct gacactcctc cactcccccg gggaactcct ccacccgtct tcacccctcc
     2401 actcccaaag ggcaccccgc cgctgactcc cagtgactca ccccagacca gaacagcatc
     2461 tggagctgtg gatgaggacg cactgactct agaagaactt gaagaacagc agaggcggat
     2521 ctgggcagct cttgagcagg ccgagagcgt aaacagcgac tccgacgttc ctgtggacac
     2581 acctttaact ggcaattccg ttgcctcatc accttgtcca aatgagctag acctccctgt
     2641 cccggaggga aaaacatctg aaaagcagac gctggatgag cctgaggtac cagagatttt
     2701 tacaaagaaa tcagaagctg gacatgcctc cagtccagac tctgaggtga catcactttg
     2761 tcagaaggaa aaagcagagt tggctccggt aaacactgaa ggtgcccttc ttgataatgg
     2821 cagtgtcgta ccaaactgtg acatcagcaa tgggggcagc cagaagctct ttcctgcaga
     2881 caccagtcct tcaacggcca ctaaaattca tagccctata cctgacatga gcaaatttgc
     2941 aactggaatc acgccatttg aatttgagaa tatggcagaa tctactggaa tgtacctcag
     3001 gataagaagc ttgttaaaga actcaccccg aaaccagcag aaaaacaaaa aggcctctga
     3061 actcgaggac tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga
     3121 tgacgatgac aagtagtgag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     3181 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3241 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3301 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3361 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     3421 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3481 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3541 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3601 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3661 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3721 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3781 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3841 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     3901 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3961 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4021 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4081 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4141 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4201 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     4261 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4321 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4381 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4441 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     4501 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4561 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4621 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4681 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4741 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4801 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4861 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4921 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4981 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     5041 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     5101 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     5161 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5221 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     5281 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5341 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     5401 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     5461 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5521 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5581 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5641 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5701 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5761 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5821 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5881 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5941 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6001 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6061 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6121 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6181 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6241 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6301 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt
     6361 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6421 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6481 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6541 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6601 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6661 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6721 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6781 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6841 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6901 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6961 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     7021 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7081 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     7141 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     7201 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7261 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7321 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7381 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7441 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7501 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7561 cacctgacgt c
//
