LOCUS       V023427                 8123 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023427
VERSION     V023427
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        4..383
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        386..661
                     /label="chicken β-actin promoter"
     intron          662..1674
                     /note="chimera between introns from chicken β-actin and
                     rabbit β-globin"
                     /label="chimeric intron"
     regulatory      1681..1690
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             1696..1719
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin®, an engineered
                     form of streptavidin"
                     /transl_table=1
                     /translation="WSHPQFEK"
                     /label="Strep-Tag II"
     CDS             1720..1743
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin®, an engineered
                     form of streptavidin"
                     /transl_table=1
                     /translation="WSHPQFEK"
                     /label="Strep-Tag II"
     CDS             1744..1815
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags"
                     /transl_table=1
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
                     /label="3xFLAG"
     CDS             1816..1836
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /transl_table=1
                     /translation="ENLYFQS"
                     /label="TEV site"
     CDS             1852..5118
                     /label="PIGO(NM_032634)"
                     /note="PIGO(NM_032634)"
                     /gene="PIGO"
     polyA_signal    5210..5265
                     /note="rabbit&nbsp;β-globin polyadenylation signal (Gil and
                     Proudfoot, 1987)"
                     /label="β-globin poly(A) signal"
     primer_bind     complement(5626..5642)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5650..5666
                     /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(5674..5704)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5719..5740
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        5799..5994
                     /note="SV40 early promoter"
                     /label="SV40 promoter"
     polyA_signal    6000..6134
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(6373..6961)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7132..7992)
                     /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(7993..8097)
                     /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 tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
     1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     1381 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
     1441 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1621 ggcggctcta gtgcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1681 gccaccatgg ctagctggag ccacccgcag ttcgagaagt ggagccaccc gcagttcgag
     1741 aaggactaca aggacgatga tgacaaggac tacaaggacg atgatgacaa ggactacaag
     1801 gacgatgatg acaaggaaaa cctgtacttc caatccgcga tcgctctcga gatgcagaaa
     1861 gcctcagtgt tgctcttcct ggcctgggtc tgcttcctct tctacgctgg cattgccctc
     1921 ttcaccagtg gcttcctgct cacccgtttg gagctcacca accatagcag ctgccaagag
     1981 cccccaggcc ctgggtccct gccatggggg agccaaggga aacctggggc ctgctggatg
     2041 gcttcccgat tttcgcgggt tgtgttggtg ctgatagatg ctctgcgatt tgacttcgcc
     2101 cagccccagc attcacacgt gcctagagag cctcctgtct ccctaccctt cctgggcaaa
     2161 ctaagctcct tgcagaggat cctggagatt cagccccacc atgcccggct ctaccgatct
     2221 caggttgacc ctcctaccac caccatgcag cgcctcaagg ccctcaccac tggctcactg
     2281 cctaccttta ttgatgctgg tagtaacttc gccagccacg ccatagtgga agacaatctc
     2341 attaagcagc tcaccagtgc aggaaggcgt gtagtcttca tgggagatga tacctggaaa
     2401 gaccttttcc ctggtgcttt ctccaaagct ttcttcttcc catccttcaa tgtcagagac
     2461 ctagacacag tggacaatgg catcctggaa cacctctacc ccaccatgga cagtggtgaa
     2521 tgggacgtgc tgattgctca cttcctgggt gtggaccact gtggccacaa gcatggccct
     2581 caccaccctg aaatggccaa gaaacttagc cagatggacc aggtgatcca gggacttgtg
     2641 gagcgtctgg agaatgacac actgctggta gtggctgggg accatgggat gaccacaaat
     2701 ggagaccatg gaggggacag tgagctggag gtctcagctg ctctctttct gtatagcccc
     2761 acagcagtct tccccagcac cccaccagag gagccagagg tgattcctca agttagcctt
     2821 gtgcccacgc tggccctgct gctgggcctg cccatcccat ttgggaatat cggggaagtg
     2881 atggctgagc tattctcagg gggtgaggac tcccagcccc actcctctgc tttagcccaa
     2941 gcctcagctc tccatctcaa tgctcagcag gtgtcccgat ttcttcatac ctactcagct
     3001 gctactcagg accttcaagc taaggagctt catcagctgc agaacctctt ctccaaggcc
     3061 tctgctgact accagtggct tctccagagc cccaaggggg ctgaggcgac actgccgact
     3121 gtgattgctg agctgcagca gttcctgcgg ggagctcggg ccatgtgcat cgagtcttgg
     3181 gctcgtttct ctctggtccg catggcgggg ggtactgctc tcttggctgc ttcctgcttt
     3241 atctgcctgc tggcatctca gtgggcaata tccccaggct ttccattctg ccctctactc
     3301 ctgacacctg tggcctgggg cctggttggg gccatagcgt atgctggact cctgggaact
     3361 attgagctga agctagatct agtgcttcta ggggctgtgg ctgcagtgag ctcattcctc
     3421 ccttttctgt ggaaagcctg ggctggctgg gggtccaaga ggcccctggc aaccctgttt
     3481 cccatccctg ggcccgtcct gttactcctg ctgtttcgct tggctgtgtt cttctctgat
     3541 agttttgttg tagctgaggc cagggccacc cccttccttt tgggctcatt catcctgctc
     3601 ctggttgtcc agcttcactg ggagggccag ctgcttccac ctaagctact cacaatgccc
     3661 cgccttggca cttcagccac aacaaacccc ccacggcaca atggtgcata tgccctgagg
     3721 cttggaattg ggttgctttt atgtacaagg ctagctgggc tttttcatcg ttgccctgaa
     3781 gagacacctg tttgccactc ctctccctgg ctgagtcctc tggcatccat ggtgggtggt
     3841 cgagccaaga atttgtggta tggagcttgt gtggcggcgc tggtggccct gttagctgcc
     3901 gtgcgcttgt ggcttcgccg ctatggtaat ctcaagagcc ccgagccacc catgctcttt
     3961 gtgcgctggg gactgcccct aatggcattg ggtactgctg cctactgggc attggcgtcg
     4021 ggggcagatg aggctccccc ccgtctccgg gtcctggtct ctggggcatc catggtgctg
     4081 cctcgggctg tagcagggct ggctgcttca gggctcgcgc tgctgctctg gaagcctgtg
     4141 acagtgctgg tgaaggctgg ggcaggcgct ccaaggacca ggactgtcct cactcccttc
     4201 tcaggccccc ccacttctca agctgacttg gattatgtgg tccctcaaat ctaccgacac
     4261 atgcaggagg agttccgggg ccggttagag aggaccaaat ctcagggtcc cctgactgtg
     4321 gctgcttatc agttggggag tgtctactca gctgctatgg tcacagccct caccctgttg
     4381 gccttcccac ttctgctgtt gcatgcggag cgcatcagcc ttgtgttcct gcttctgttt
     4441 ctgcagagct tccttctcct acatctgctt gctgctggga tacccgtcac cacccctggt
     4501 ccttttactg tgccatggca ggcagtctcg gcttgggccc tcatggccac acagaccttc
     4561 tactccacag gccaccagcc tgtctttcca gccatccatt ggcatgcagc cttcgtggga
     4621 ttcccagagg gtcatggctc ctgtacttgg ctgcctgctt tgctagtggg agccaacacc
     4681 tttgcctccc acctcctctt tgcagtaggt tgcccactgc tcctgctctg gcctttcctg
     4741 tgtgagagtc aagggctgcg gaagagacag cagcccccag ggaatgaagc tgatgccaga
     4801 gtcagacccg aggaggaaga ggagccactg atggagatgc ggctccggga tgcgcctcag
     4861 cacttctatg cagcactgct gcagctgggc ctcaagtacc tctttatcct tggtattcag
     4921 attctggcct gtgccttggc agcctccatc cttcgcaggc atctcatggt ctggaaagtg
     4981 tttgccccta agttcatatt tgaggctgtg ggcttcattg tgagcagcgt gggacttctc
     5041 ctgggcatag ctttggtgat gagagtggat ggtgctgtga gctcctggtt caggcagcta
     5101 tttctggccc agcagaggta gaattcgcgg ccgcatctag aagatctttt tccctctgcc
     5161 aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa
     5221 tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat
     5281 gggagggcaa atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat
     5341 gcccatatgc tggctgccat gaacaaaggt tggctataaa gaggtcatca gtatatgaaa
     5401 cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt
     5461 tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt
     5521 ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc
     5581 ttatggagat ccctcgacct gcagcccaag cttggcgtaa tcatggtcat agctgtttcc
     5641 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5701 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5761 cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc
     5821 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     5881 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     5941 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagcta
     6001 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     6061 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     6121 atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6181 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6241 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6301 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6361 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6421 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6481 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6541 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     6601 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6661 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6721 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6781 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     6841 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6901 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6961 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7021 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7081 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7141 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7201 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7261 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7321 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7381 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7441 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7501 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7561 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7621 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7681 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7741 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7801 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7861 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7921 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7981 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8041 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8101 atttccccga aaagtgccac ctg
//
