LOCUS       V042189                 8710 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042189
VERSION     V042189
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..3366
                     /label="Kif20a(NM_001166406)"
                     /note="Kif20a(NM_001166406)"
                     /gene="Kif20a"
     CDS             3373..3462
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    3486..4074
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4140..4188
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4390..4818
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4832..5161
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5228..6022
                     /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    6198..6319
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6368..6384)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6392..6408
                     /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(6416..6446)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6461..6482
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6770..7358)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7529..8389)
                     /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(8390..8494)
                     /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 ccaccatgtc tcaccggatc
      721 ctttctccgc cagcgggctt actctctgat gaggatgttg tagactctcc catattagaa
      781 tccacagctg cagatctgag atctgttgtc cgaaaggacc tgttgtcaga ctgctctgtc
      841 atctctgcct ctctggagga caagcaggct ctactggagg acacctcaga gaaggtgaaa
      901 gtttaccttc gaattcgacc attcttgact tcagagttgg accgacagga agatcagggc
      961 tgcgtctgca ttgagaatac agaaaccctt gtgctgcagg cacccaaaga ctcctttgcc
     1021 ttgaagagta atgagagggg agttggtcag gccactcaca aatttacctt ttcccagata
     1081 tttgggccag aagtgggaca ggtggccttc ttcaatctga ccatgaagga gatggtgaag
     1141 gatgtgctca aagggcagaa ctggctcatc tacacctatg gagtcaccaa ctcggggaaa
     1201 acgtacacca ttcaaggtac tagtaaggat gcggggatcc tgccccagtc cctggctctc
     1261 atctttaata gtctccaggg ccagcttcat ccgacacctg acctgaagcc cttattgtcc
     1321 aatgaagtaa tctggctaga cagcaagcag attcgacagg aggaaatgaa gaagctgtcc
     1381 ttgctaattg gaggcctcca agaggaggag ctgtccacct ccgtgaagaa acgtgtccac
     1441 actgaaagtc ggataggtgc cagcaacagc tttgacagtg gcgttgctgg gctgtcttcc
     1501 accagtcagt tcaccagcag cagccagcta gatgaaacaa gtcaattatg ggcacaacca
     1561 gacactgtcc cagtaagtgt tccagcagac attcgcttct ctgtctggat ctccttcttt
     1621 gagatctaca atgaattgct ttatgacctg ttagaaccac ctagccatca gcataagaga
     1681 cagacacttc ggctgtgtga ggatcagaac ggcaatcctt acgtgaaaga tctcaattgg
     1741 attcatgttc gtgatgttga ggaagcctgg aaactattga aagtgggtcg caagaaccag
     1801 agctttgcca gcacccacat gaaccagcaa tccagccgca gtcacagcat cttctcaatc
     1861 agaatcttgc accttcaggg agaaggggat atagtgccca agatcagtga actgtcactc
     1921 tgtgacctgg ctggctctga gcgctgcaaa catcaaaaaa gtggtgagcg gctaaaggag
     1981 gcagggaaca ttaacacttc tctgcacacc ctgggccgct gtattgctgc cctgcgacag
     2041 aatcagcaga accggtcaaa gcagaacctg attcctttcc gtgacagcaa gttgactcgt
     2101 gtgttccaag gcttcttcac aggtcgaggt cgttcctgta tgattgtcaa tgtgaatccc
     2161 tgtgcatcta catatgacga gactcttcat gcagccaaat tctcagccct agccagccag
     2221 cttgtgcacg ccccacctgt gcatctggga atcccatccc tgcattcatt catcaagaag
     2281 cacagtcctc aggttggccc tggcttagag aaagaggaca aggccgactc agaccttgag
     2341 gacagtcctg aagacgaagc tgacgtctcc gtgtatggca aggaggaact actccaggtg
     2401 gtggaagcca tgaaagcgct acttttaaaa gaacgacaag aaaagctgca gttggagata
     2461 cagctccgag aggaaatttg caacgaaatg gtggaacaga tgcaacagcg ggagcagtgg
     2521 tgcagtgagc gtttggacaa ccaaaaggaa ctgatggagg aactgtacga ggagaaactg
     2581 aagatcctga aggaatcact gacaactttc taccaagagc agatccagga gcgggatgaa
     2641 aaaattgaag agctagaaac tctgttgcag gaagccaaac agcagccagc ggcacaacag
     2701 tcaggggggc tgtccctcct ccggcggtca cagaggctgg cagcgtctgc ctccactcag
     2761 cagttccagg aagttaaagc tgaactagag caatgcaaaa cagagttaag ctctaccact
     2821 gcagagctgc acaagtatca gcaagtgtta aaaccgccac ccccagccaa gcctttcacc
     2881 attgatgtgg ataagaagtt agaggagggc cagaagaata taagactgct acggacagag
     2941 ctccagaaac taggacagtc tctgcagtca gcagaaagag cctgttgcca cagcaccgga
     3001 gcaggaaaac ttcgtcaagc attaaccaac tgtgatgaca tcttaatcaa gcagaaccag
     3061 accctggctg agctgcagaa taacatggtg ctagtgaaac tggaccttca gaaaaaggca
     3121 gcctgcattg ctgagcagta tcacactgtg ctaaagctcc aaggccaagc ctctgccaaa
     3181 aagcggctgg gagcaaacca ggaaaaccag caaccaaatc atcaaccccc aggaaagaaa
     3241 ccattcctgc gaaacttact tccccgaaca cctacctgcc aaagctcaac agacagcagc
     3301 ccttatgcgc ggatcttgcg ctcacggcac tctcctttac tcaaatctcc ctttggcaaa
     3361 aaatacctcg aggaacaaaa actaatatca gaggaagatt tggaacagaa actaatcagt
     3421 gaagaagatt tagagcaaaa gttaatttcc gaggaggact tatagtgagg gcccgatatc
     3481 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     3541 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     3601 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     3661 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     3721 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     3781 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     3841 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
     3901 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     3961 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     4021 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa
     4081 acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     4141 ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg
     4201 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     4261 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     4321 cagccaacgt cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg
     4381 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4441 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4501 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt
     4561 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     4621 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4681 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     4741 ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac
     4801 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     4861 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     4921 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4981 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     5041 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     5101 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     5161 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     5221 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     5281 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     5341 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     5401 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     5461 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     5521 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     5581 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     5641 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     5701 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     5761 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     5821 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     5881 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     5941 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     6001 ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga
     6061 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     6121 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     6181 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     6241 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     6301 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     6361 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     6421 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     6481 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     6541 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     6601 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6661 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6721 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6781 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6841 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6901 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6961 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     7021 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     7081 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     7141 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     7201 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     7261 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     7321 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     7381 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     7441 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     7501 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     7561 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     7621 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     7681 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7741 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7801 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7861 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7921 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7981 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     8041 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     8101 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     8161 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     8221 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     8281 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     8341 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     8401 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     8461 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     8521 acgtcgacgg atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc
     8581 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
     8641 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
     8701 tctgcttagg
//
