LOCUS       V020273                10144 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020273
VERSION     V020273
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..5623
                     /label="ZC3H13(NM_015070)"
                     /note="ZC3H13(NM_015070)"
                     /gene="ZC3H13"
     CDS             5645..5710
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    5739..5963
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      6009..6437
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6451..6780
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6847..7641
                     /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    7817..7938
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7987..8003)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8011..8027
                     /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(8035..8065)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8080..8101
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8389..8977)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9148..10008)
                     /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(10009..10113)
                     /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 catgtcaaaa attagaagga aggtcacagt
      961 ggaaaatacc aagactatat ctgatagcac atcccgaaga cccagtgtat ttgagaggct
     1021 tggacccagc actggcagta cagcagagac acagtgccgt aactggctga agactggcaa
     1081 ctgcctctat ggaaacacat gtagattcgt acatggccct tcacctcgtg gtaaaggtta
     1141 tagcagcaat tatagaaggt caccagaaag acctacaggg gatcttagag aaagaatgaa
     1201 gaacaagcgc caagacgtgg acactgagcc ccagaaacga aatacagagg agtcatcctc
     1261 acctgttagg aaagaatctt caagagggag acatagggaa aaggaagaca taaaaatcac
     1321 taaggaaaga actccagaaa gtgaagaaga aaatgtagaa tgggaaacta atagagatga
     1381 ttctgacaat ggagatatta attatgatta tgttcatgaa ttgtcattgg aaatgaagcg
     1441 tcagaagata cagagggaat taatgaagct ggaacaagaa aacatggaga agagagaaga
     1501 aattatcatt aaaaaggagg tttcaccaga agtggttaga tcaaaattgt ccccgtcacc
     1561 ttctctaaga aagtctagca aatctccgaa gcgaaaatca agcccgaagt cgtcttcagc
     1621 tagcaagaaa gataggaaga catctgcagt atcttctccc ctgttggacc agcagagaaa
     1681 ttcaaaaacc aaccaaagta aaaagaaagg accacgtact cctagtccac cccctcctat
     1741 accagaagat atcgctctgg ggaaaaaata caaagaaaaa tataaagtaa aagacaggat
     1801 agaagaaaaa acaagagatg gaaaggacag aggacgagat tttgaacgac aaagagaaaa
     1861 gagagacaag ccaaggtcta cttccccagc aggacagcat cattctccta tatcttctag
     1921 acatcactca tcttcctcac aatcaggatc atctattcaa agacattctc cttctcctcg
     1981 tcgaaaaaga actccttcac catcttatca gcggacacta actccacctt tacgacgctc
     2041 tgcctctcct tatccttcac attctttgtc gtctccccag agaaagcaga gtcctccaag
     2101 acatcgctct ccaatgcgag agaaagggag acatgatcat gaacgaactt cacagtctca
     2161 tgatcgacgc cacgaaagga gggaagatac taggggcaaa cgagacagag aaaaggactc
     2221 aagagaagaa cgagaatatg aacaggatca gagctcttct agagaccaca gagatgacag
     2281 agaacctcga gatggtcggg atcggagaga tgccagagat actagggacc gaagggaact
     2341 aagagactcc agagacatgc gggactcaag ggagatgaga gattatagca gagataccaa
     2401 agagagccgt gatcccagag attctcggtc cactcgtgat gcccatgact acagggaccg
     2461 tgaaggtcga gatactcatc gaaaggagga tacatatcca gaagaatccc ggagttatgg
     2521 ccgaaaccat ttgagagaag aaagttctcg tacggaaata aggaatgagt ccagaaatga
     2581 gtctcgaagt gaaattagaa atgaccgaat gggccgaagt agggggaggg ttcctgagtt
     2641 acctgaaaag ggaagtcgag gctcaagagg ttctcaaatt gatagtcaca gtagtaatag
     2701 caactatcat gacagctggg aaactcgaag tagctatcct gaaagagata gatatcctga
     2761 aagagacaac agagatcaag caagggattc ttcctttgag agaagacatg gagagcgaga
     2821 ccgtcgtgac aacagagaga gagatcaaag accaagctca ccaattcgac atcagggaag
     2881 gaatgacgag cttgagcgtg atgaaagaag agaggaacga agagtagaca gagtggatga
     2941 taggagagat gaaagggcta gagagagaga tcgggaacga gaacgagaca gggagcggga
     3001 gagagagagg gaacgtgaac gggatcggga aagagaaaaa gagagagaac tagaaagaga
     3061 gcgtgctagg gaacgggaga gagaaagaga aaaagagaga gatcgtgaaa gggatagaga
     3121 ccgagaccac gatcgagagc gggaaagaga gagggaacga gacagggaaa aagaacggga
     3181 acgagaaaga gaagagagag agagggagag agagcgagaa cgggagagag agcgagagcg
     3241 agaacgggaa cgagaaagag cgagagaaag ggataaagaa cgagaacgcc aaagggattg
     3301 ggaagacaaa gacaaaggac gagatgaccg cagagaaaag cgagaagaga tccgagaaga
     3361 taggaatcca agagatggac atgatgaaag aaaatcaaag aagcgctata gaaatgaagg
     3421 gagtcccagc cctagacagt ccccgaagcg ccggcgtgaa cattctccgg acagtgatgc
     3481 ctacaacagt ggagatgata aaaatgaaaa acacagactc ttgagccaag ttgtacgacc
     3541 tcaagaatct cgttctctta gtccctcgca cctcacagaa gacagacagg gtagatggaa
     3601 agaggaggat cgtaaaccag aaaggaaaga gagttcaagg cgctacgaag aacaggaact
     3661 caaggagaaa gtttcttctg tagataaaca gagagaacag acagaaatcc tggaaagctc
     3721 aagaatgcgt gcacaggaca ttataggaca ccaccagtct gaagatcgag agacatctga
     3781 tcgagctcat gatgaaaaca agaagaaagc aaaaattcaa aagaaaccaa ttaagaaaaa
     3841 gaaagaggat gatgttggaa tagagagggg taacatagag acaacatctg aagatggtca
     3901 agtattttca ccaaaaaaag gacagaaaaa gaaaagcatt gaaaaaaaac gtaaaaaatc
     3961 caaaggtgat tctgatattt ctgatgaaga agcagcccag caaagtaaga agaaaagagg
     4021 cccacggact ccccctataa caactaaaga ggaattggtt gaaatgtgca atggtaagaa
     4081 tggtattcta gaggactccc agaaaaaaga agatacagca ttcagtgact ggtctgatga
     4141 ggatgtccct gaccgtacag aggtgacaga agcagagcat actgccaccg ccacgactcc
     4201 tggtagtacc ccttctcctc tatcttctct tcttcctcct ccaccgcctg tggctactgc
     4261 cactgctaca actgtgcctg caactcttgc tgccactact gctgctgccg ccacctcttt
     4321 cagcacatct gccatcacta tttccacctc tgccaccccc accaatacca ccaataatac
     4381 ttttgccaat gaagactcac acagaaaatg ccacagaaca cgagtagaaa aagtagagac
     4441 gcctcacgtg actatagaag atgcacagca tcgcaagcct atggatcaaa agaggagcag
     4501 cagcctcggg agcaatcgga gtaaccgtag tcatacgtct ggtcgtcttc gctccccatc
     4561 caatgattca gcccatcgaa gtggagatga ccaaagtggt cgaaagagag tactgcacag
     4621 tggctcaaga gatagagaaa aaacaaaaag cctggaaatc acaggagaga gaaaatctag
     4681 gattgatcag ttaaagcgtg gagaacccag tcgaagtact tcttcagatc gccaggattc
     4741 aagaagccat agttcaagaa gaagttctcc agagtcagat cgacaggtcc attcaagatc
     4801 tgggtcattt gatagcagag acaggcttca agaacgagat cgatatgaac acgacagaga
     4861 gcgcgagaga gagaggagag atacgaggca gagagaatgg gaccgagatg ctgataaaga
     4921 ttggccacgc aacagggatc gagatagatt gcgagaacga gaacgagaga gagaacgaga
     4981 caaaaggaga gacttggata gggaaagaga gagactaatt tctgattctg ttgaaaggga
     5041 cagggacaga gacagagaca gaacttttga gagttctcaa atagagtctg tgaaacgctg
     5101 tgaagcaaaa ctggaaggtg aacatgaaag ggatctagaa agcacttccc gagactctct
     5161 agccttggat aaagagagaa tggataaaga tctgggatct gtgcagggat ttgaagaaac
     5221 aaataaatcc gagagaactg agagtctgga agcaggagat gacgagtcca agttagatga
     5281 tgcacattca ttaggctctg gtgctggaga aggatacgag ccaatcagtg atgacgaact
     5341 agatgaaatt ctggcaggtg atgcagaaaa gagggaggac caacaggatg aggagaagat
     5401 gccagatccc ttagatgtga tagatgtgga ttggtctggt cttatgccaa agcatccaaa
     5461 agaaccacga gagcctgggg ctgcactctt aaaattcaca cctggagctg ttatgctaag
     5521 agttgggatt tctaaaaagt tggcaggttc tgaactcttt gccaaagtca aagaaacatg
     5581 tcagagactt ttagaaaaac ccaaaggtag tttcatttta cttctcgagt ctagagggcc
     5641 cttcgactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga
     5701 cgatgacaag tgagtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     5761 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     5821 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     5881 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     5941 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     6001 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     6061 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     6121 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     6181 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     6241 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     6301 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     6361 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     6421 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     6481 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     6541 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     6601 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     6661 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     6721 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     6781 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     6841 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     6901 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     6961 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     7021 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     7081 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     7141 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     7201 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     7261 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     7321 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     7381 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     7441 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     7501 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     7561 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     7621 cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac
     7681 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     7741 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     7801 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     7861 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     7921 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     7981 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     8041 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     8101 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     8161 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     8221 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     8281 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     8341 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     8401 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     8461 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     8521 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     8581 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     8641 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     8701 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     8761 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     8821 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     8881 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     8941 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     9001 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     9061 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     9121 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     9181 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     9241 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     9301 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     9361 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     9421 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     9481 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     9541 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     9601 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     9661 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     9721 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     9781 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     9841 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     9901 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     9961 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
    10021 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
    10081 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
    10141 cgtc
//
