LOCUS       V032029                 6558 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032029
VERSION     V032029
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        4..383
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        385..662
                     /label="chicken β-actin promoter"
     intron          663..1679
                     /note="chimera between introns from chicken β-actin and
                     rabbit β-globin"
                     /label="chimeric intron"
     CDS             1740..3539
                     /label="KLHL24(NM_017644)"
                     /note="KLHL24(NM_017644)"
                     /gene="KLHL24"
     CDS             3540..3563
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     polyA_signal    3646..3701
                     /note="rabbit&nbsp;β-globin polyadenylation signal (Gil and
                     Proudfoot, 1987)"
                     /label="β-globin poly(A) signal"
     primer_bind     complement(4062..4078)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4086..4102
                     /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(4110..4140)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4154..4175
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        4234..4429
                     /note="SV40 early promoter"
                     /label="SV40 promoter"
     polyA_signal    4435..4569
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(4807..5395)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5566..6426)
                     /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(6427..6531)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg ggggggcccc ccccaggcgg ggcggggcgg ggcgaggggc
      541 ggggcggggc gaggcggaaa ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgt tgcgcgctgc cttccccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttcgccacca
     1741 tggtactaat attgggacgc agactaaaca gagaggatct tggggtgcgt gattccccag
     1801 caactaagcg aaaagttttt gaaatggacc ccaaatctct gacaggtcat gagttttttg
     1861 acttctcttc aggatcatcc catgccgaaa acatactcca gatatttaat gaatttcgtg
     1921 atagccgctt attcacagat gttatcattt gtgtggaagg aaaagaattt ccttgccata
     1981 gagctgtgct ctcagcctgt agcagctact tcagagctat gttttgtaat gaccacaggg
     2041 aaagccgaga aatgttggtt gagatcaatg gtattttagc tgaagctatg gaatgttttt
     2101 tgcagtatgt ttatactgga aaggtgaaga tcactacaga gaatgtacag tatctctttg
     2161 agacatcaag cctctttcag attagtgttc tccgtgatgc atgtgccaag ttcttggagg
     2221 agcaacttga tccttgtaat tgcttaggaa tccagcgctt tgctgatacc cattcactca
     2281 aaacactctt cacaaaatgc aaaaattttg cgttacagac ttttgaggat gtatcccagc
     2341 acgaagaatt tcttgagctt gacaaagatg aacttattga ttatatttgt agtgatgaac
     2401 ttgttattgg taaagaggag atggtttttg aagccgtcat gcgttgggtc tatcgtgccg
     2461 ttgatctgag aagaccactg ttacacgagc tcctgacaca tgtgagactc cctctgttgc
     2521 atcccaacta ctttgttcaa acagttgaag tggaccaatt gatccagaat tctcctgagt
     2581 gttatcagtt gttgcatgaa gcaagacggt accacatact tgggaatgaa atgatgtccc
     2641 caaggactag gccacgcagg tccactggct attctgaggt gatagttgtc gttggaggat
     2701 gtgagcgagt tggaggattt aatcttccat acactgagtg ctacgatcct gtaacaggag
     2761 aatggaagtc tttggctaag cttccagaat ttaccaaatc agagtatgca gtctgtgctc
     2821 taaggaatga cattcttgtt tcaggtggaa gaatcaacag ccgtgatgtc tggatttata
     2881 actcacagtt aaatatttgg atcagagttg cctctctcaa taaaggcaga tggcgtcaca
     2941 aaatggctgt cctccttggt aaagtatatg ttgtcggtgg ctatgatggg caaaacagac
     3001 ttagcagcgt agaatgttat gattcctttt caaatcgatg gactgaagtt gctcccctta
     3061 aggaagccgt gagttctcct gcagtgacta gctgtgtagg caaactgttt gtgattggtg
     3121 gaggacctga tgataatact tgttctgata aggttcaatc ttatgatcca gaaaccaatt
     3181 cttggctact tcgtgcagct atcccaattg ccaaaaggtg tataacagct gtatccctaa
     3241 acaacctgat ctatgttgcc ggtggactga ccaaggcaat atactgttac gatccagttg
     3301 aagattactg gatgcacgta cagaatacat tcagccgtca ggaaaactgt ggtatgtctg
     3361 tgtgtaatgg taaaatatat atcctgggcg gaagacggga aaatggagaa gccacagaca
     3421 ctattctctg ttatgatcct gcaacaagta tcatcacagg ggtagctgca atgcccaggc
     3481 cagtgtccta tcatggctgt gtgactattc atagatacaa tgagaaatgc tttaaactcg
     3541 attacaagga tgacgacgat aagtgactcg agctagcaga tctttttccc tctgccaaaa
     3601 attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta
     3661 ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga
     3721 gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc
     3781 atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc
     3841 cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta
     3901 tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac
     3961 tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat
     4021 ggagatccct cgacctgcag cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg
     4081 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     4141 gcctgggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     4201 tccagtcggg aaacctgtcg tgccagcgga tccgcatctc aattagtcag caaccatagt
     4261 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4321 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     4381 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctaacttg
     4441 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4501 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     4561 gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4621 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4681 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg ataacgcagg
     4741 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4801 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4861 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4921 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4981 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5041 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5101 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5161 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5221 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5281 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5341 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5401 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5461 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5521 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5581 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5641 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5701 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5761 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5821 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5881 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5941 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6001 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6061 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6121 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6181 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6241 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6301 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6361 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6421 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6481 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6541 ccgaaaagtg ccacctgg
//
