LOCUS       V028466                 7685 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028466
VERSION     V028466
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..3175
                     /label="CHUK(NM_001278)"
                     /note="CHUK(NM_001278)"
                     /gene="CHUK"
     CDS             3182..3247
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3285..3509
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3555..3983
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3997..4326
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4393..5187
                     /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    5361..5482
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5531..5547)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5555..5571
                     /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(5579..5609)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5624..5645
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5933..6518)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6689..7549)
                     /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(7550..7654)
                     /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 atggagcggc ccccggggct
      961 gcggccgggc gcgggcgggc cctgggagat gcgggagcgg ctgggcaccg gcggcttcgg
     1021 gaacgtctgt ctgtaccagc atcgggaact tgatctcaaa atagcaatta agtcttgtcg
     1081 cctagagcta agtaccaaaa acagagaacg atggtgccat gaaatccaga ttatgaagaa
     1141 gttgaaccat gccaatgttg taaaggcctg tgatgttcct gaagaattga atattttgat
     1201 tcatgatgtg cctcttctag caatggaata ctgttctgga ggagatctcc gaaagctgct
     1261 caacaaacca gaaaattgtt gtggacttaa agaaagccag atactttctt tactaagtga
     1321 tatagggtct gggattcgat atttgcatga aaacaaaatt atacatcgag atctaaaacc
     1381 tgaaaacata gttcttcagg atgttggtgg aaagataata cataaaataa ttgatctggg
     1441 atatgccaaa gatgttgatc aaggaagtct gtgtacatct tttgtgggaa cactgcagta
     1501 tctggcccca gagctctttg agaataagcc ttacacagcc actgttgatt attggagctt
     1561 tgggaccatg gtatttgaat gtattgctgg atataggcct tttttgcatc atctgcagcc
     1621 atttacctgg catgagaaga ttaagaagaa ggatccaaag tgtatatttg catgtgaaga
     1681 gatgtcagga gaagttcggt ttagtagcca tttacctcaa ccaaatagcc tttgtagttt
     1741 agtagtagaa cccatggaaa actggctaca gttgatgttg aattgggacc ctcagcagag
     1801 aggaggacct gttgacctta ctttgaagca gccaagatgt tttgtattaa tggatcacat
     1861 tttgaatttg aagatagtac acatcctaaa tatgacttct gcaaagataa tttcttttct
     1921 gttaccacct gatgaaagtc ttcattcact acagtctcgt attgagcgtg aaactggaat
     1981 aaatactggt tctcaagaac ttctttcaga gacaggaatt tctctggatc ctcggaaacc
     2041 agcctctcaa tgtgttctag atggagttag aggctgtgat agctatatgg tttatttgtt
     2101 tgataaaagt aaaactgtat atgaagggcc atttgcttcc agaagtttat ctgattgtgt
     2161 aaattatatt gtacaggaca gcaaaataca gcttccaatt atacagctgc gtaaagtgtg
     2221 ggctgaagca gtgcactatg tgtctggact aaaagaagac tatagcaggc tctttcaggg
     2281 acaaagggca gcaatgttaa gtcttcttag atataatgct aacttaacaa aaatgaagaa
     2341 cactttgatc tcagcatcac aacaactgaa agctaaattg gagttttttc acaaaagcat
     2401 tcagcttgac ttggagagat acagcgagca gatgacgtat gggatatctt cagaaaaaat
     2461 gctaaaagca tggaaagaaa tggaagaaaa ggccatccac tatgctgagg ttggtgtcat
     2521 tggatacctg gaggatcaga ttatgtcttt gcatgctgaa atcatggagc tacagaagag
     2581 cccctatgga agacgtcagg gagacttgat ggaatctctg gaacagcgtg ccattgatct
     2641 atataagcag ttaaaacaca gaccttcaga tcactcctac agtgacagca cagagatggt
     2701 gaaaatcatt gtgcacactg tgcagagtca ggaccgtgtg ctcaaggagc tgtttggtca
     2761 tttgagcaag ttgttgggct gtaagcagaa gattattgat ctactcccta aggtggaagt
     2821 ggccctcagt aatatcaaag aagctgacaa tactgtcatg ttcatgcagg gaaaaaggca
     2881 gaaagaaata tggcatctcc ttaaaattgc ctgtacacag agttctgccc ggtcccttgt
     2941 aggatccagt ctagaaggtg cagtaacccc tcagacatca gcatggctgc ccccgacttc
     3001 agcagaacat gatcattctc tgtcatgtgt ggtaactcct caagatgggg agacttcagc
     3061 acaaatgata gaagaaaatt tgaactgcct tggccattta agcactatta ttcatgaggc
     3121 aaatgaggaa cagggcaata gtatgatgaa tcttgattgg agttggttaa cagaactcga
     3181 ggactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga
     3241 tgacaagtag tgagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     3301 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     3361 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     3421 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     3481 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     3541 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3601 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3661 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3721 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3781 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3841 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     3901 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     3961 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     4021 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4081 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4141 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4201 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4261 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4321 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     4381 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4441 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4501 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4561 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4621 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4681 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4741 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4801 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4861 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4921 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4981 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5041 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5101 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5161 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5221 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5281 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5341 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     5401 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     5461 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     5521 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     5581 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     5641 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5701 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     5761 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5821 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5881 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5941 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6001 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6061 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6121 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6181 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6241 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6301 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6361 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6421 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt
     6481 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6541 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6601 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6661 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6721 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6781 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6841 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6901 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6961 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7021 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7081 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7141 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7201 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7261 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7321 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7381 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7441 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7501 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7561 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7621 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7681 acgtc
//
