LOCUS       V043901                 7252 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043901
VERSION     V043901
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             925..3003
                     /label="GMPS(NM_003875)"
                     /note="GMPS(NM_003875)"
                     /gene="GMPS"
     CDS             3016..3081
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3116..3340
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3386..3814
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3828..4157
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4169..4768
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    4928..5049
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5098..5114)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5122..5138
                     /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(5146..5176)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5191..5212
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5500..6085)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6256..7116)
                     /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(7117..7221)
                     /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 aagctttcta gagaattcgc caccatggct ctgtgcaacg gagactccaa gctggagaat
      961 gctggaggag accttaagga tggccaccac cactatgaag gagctgttgt cattctggat
     1021 gctggtgctc agtacgggaa agtcatagac cgaagagtga gggaactgtt cgtgcagtct
     1081 gaaattttcc ccttggaaac accagcattt gctataaagg aacaaggatt ccgtgctatt
     1141 atcatctctg gaggacctaa ttctgtgtat gctgaagatg ctccctggtt tgatccagca
     1201 atattcacta ttggcaagcc tgttcttgga atttgctatg gtatgcagat gatgaataag
     1261 gtatttggag gtactgtgca caaaaaaagt gtcagagaag atggagtttt caacattagt
     1321 gtggataata catgttcatt attcaggggc cttcagaagg aagaagttgt tttgcttaca
     1381 catggagata gtgtagacaa agtagctgat ggattcaagg ttgtggcacg ttctggaaac
     1441 atagtagcag gcatagcaaa tgaatctaaa aagttatatg gagcacagtt ccaccctgaa
     1501 gttggcctta cagaaaatgg aaaagtaata ctgaagaatt tcctttatga tatagctgga
     1561 tgcagtggaa ccttcaccgt gcagaacaga gaacttgagt gtattcgaga gatcaaagag
     1621 agagtaggca cgtcaaaagt tttggtttta ctcagtggtg gagtagactc aacagtttgt
     1681 acagctttgc taaatcgtgc tttgaaccaa gaacaagtca ttgctgtgca cattgataat
     1741 ggctttatga gaaaacgaga aagccagtct gttgaagagg ccctcaaaaa gcttggaatt
     1801 caggtcaaag tgataaatgc tgctcattct ttctacaatg gaacaacaac cctaccaata
     1861 tcagatgaag atagaacccc acggaaaaga attagcaaaa cgttaaatat gaccacaagt
     1921 cctgaagaga aaagaaaaat cattggggat acttttgtta agattgccaa tgaagtaatt
     1981 ggagaaatga acttgaaacc agaggaggtt ttccttgccc aaggtacttt acggcctgat
     2041 ctaattgaaa gtgcatccct tgttgcaagt ggcaaagctg aactcatcaa aacccatcac
     2101 aatgacacag agctcatcag aaagttgaga gaggagggaa aagtaataga acctctgaaa
     2161 gattttcata aagatgaagt gagaattttg ggcagagaac ttggacttcc agaagagtta
     2221 gtttccaggc atccatttcc aggtcctggc ctggcaatca gagtaatatg tgctgaagaa
     2281 ccttatattt gtaaggactt tcctgaaacc aacaatattt tgaaaatagt agctgatttt
     2341 tctgcaagtg ttaaaaagcc acatacccta ttacagagag tcaaagcctg cacaacagaa
     2401 gaggatcagg agaagctgat gcaaattacc agtctgcatt cactgaatgc cttcttgctg
     2461 ccaattaaaa ctgtaggtgt gcagggtgac tgtcgttcct acagttacgt gtgtggaatc
     2521 tccagtaaag atgaacctga ctgggaatca cttatttttc tggctaggct tatacctcgc
     2581 atgtgtcaca acgttaacag agttgtttat atatttggcc caccagttaa agaacctcct
     2641 acagatgtta ctcccacttt cttgacaaca ggggtgctca gtactttacg ccaagctgat
     2701 tttgaggccc ataacattct cagggagtct gggtatgctg ggaaaatcag ccagatgccg
     2761 gtgattttga caccattaca ttttgatcgg gacccacttc aaaagcagcc ttcatgccag
     2821 agatctgtgg ttattcgaac ctttattact agtgacttca tgactggtat acctgcaaca
     2881 cctggcaatg agatccctgt agaggtggta ttaaagatgg tcactgagat taagaagatt
     2941 cctggtattt ctcgaattat gtatgactta acatcaaagc ccccaggaac tactgagtgg
     3001 gagctcgaga ccggtgacta caaagaccat gacggtgatt ataaagatca tgacatcgat
     3061 tacaaggatg acgatgacaa gtgaccggta agtttaaacc gctgatcagc ctcgactgtg
     3121 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     3181 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     3241 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     3301 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     3361 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3421 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3481 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3541 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3601 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3661 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3721 atctcggtct attcttttga tttataaggg attttggcca tttcggccta ttggttaaaa
     3781 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     3841 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3901 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3961 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4021 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4081 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4141 gcctaggctt ttgcaaaaag ctcccgggat gaccgagtac aagcccacgg tgcgcctcgc
     4201 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     4261 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
     4321 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     4381 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     4441 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     4501 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     4561 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     4621 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     4681 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     4741 gtgcatgacc cgcaagcccg gtgcctgatt cgaaatgacc gaccaagcga cgcccaacct
     4801 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4861 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4921 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4981 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5041 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5101 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5161 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5221 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5281 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5341 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5401 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5461 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5521 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5581 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5641 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5701 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5761 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5821 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5881 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5941 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6001 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat
     6061 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6121 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6181 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6241 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6301 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6361 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6421 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6481 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6541 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6601 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6661 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6721 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6781 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6841 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6901 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6961 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7021 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7081 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7141 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7201 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
