LOCUS       V020179                 7849 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020179
VERSION     V020179
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             913..3600
                     /label="N4BP1(NM_153029)"
                     /note="N4BP1(NM_153029)"
                     /gene="N4BP1"
     CDS             3613..3678
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3713..3937
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3983..4411
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4425..4754
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4766..5365
                     /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    5525..5646
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5695..5711)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5719..5735
                     /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(5743..5773)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5788..5809
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6097..6682)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6853..7713)
                     /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(7714..7818)
                     /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 aagcttgcca ccatggcggc ccgggcggtg ctggacgagt tcactgcgcc agctgagaag
      961 gcggagctgc tggagcagag ccgcggccgt atcgagggcc tgtttggcgt gagcctagcc
     1021 gtgctcggcg cgctaggggc tgaggagccg ctgcccgcgc gcatctggct gcagctctgc
     1081 ggggcgcagg aggcggtgca cagcgccaag gaatatatta aaggaatctg tgaacctgaa
     1141 ctagaagaaa gagaatgtta ccccaaggac atgcactgca tttttgttgg ggcagagagc
     1201 ctgtttctga aaagcttgat tcaggatact tgtgctgacc tctgcattct ggacattggc
     1261 cttcttggca tcagaggaag tgctgaggct gtggtcatgg ctaggagtca cattcaacaa
     1321 tttgtaaagc tctttgaaaa taaagagaac ctacccagta gtcagaaaga atcagaggtg
     1381 aaaagggaat tcaaacaatt tgttgaagcc catgcagaca attacacaat ggatttgttg
     1441 attttgccca cttccttgaa aaaagaactt ttgacactca cacaaggtga ggagaatctc
     1501 tttgaaacag gagatgatga ggttattgaa atgagagatt ctcaacaaac agagtttaca
     1561 cagaatgctg ccacagggct gaatatttct agagatgaaa ctgttttgca ggaagaggca
     1621 agaaataaag ctgggactcc tgtttctgag cttacaaaac aaatggacac agtcctttct
     1681 agctcaccag atgtgctttt tgatccaata aatggtctaa ccccagatga agaggcactt
     1741 tccaatgaga gaatttgtca gaaaaggaga ttttctgatt ctgaagaaag gcatacgaag
     1801 aagcagtttt ctttggaaaa tgttcaggag ggggagattt tacacgatgc taagacattg
     1861 gctggaaatg taatagctga cctatctgat tcttctgctg attctgaaaa tttaagtcca
     1921 gatataaaag aaactactga ggaaatggaa tacaacatcc tcgtaaactt ttttaaaacc
     1981 atgggctact cccaagaaat tgttgaaaag gtcattaagg tgtatggacc atctactgaa
     2041 ccattattgc tcttagagga aattgaaaaa gaaaataaaa gattccaaga agacagagaa
     2101 ttttcagctg gtacagtgta tccagagacc aacaaaacca aaaataaagg tgtttatagc
     2161 agcacaaatg agcttacaac tgattccact ccaaagaaaa cacaggctca cacacagcaa
     2221 aatatggtag aaaaattttc tcagttacca ttcaaagtgg aagctaaacc atgtacctca
     2281 aattgtagaa ttaatacttt cagaacagtg ccaatagaac agaaacatga agtctggggt
     2341 tcaaaccaga actacatttg taacacagac cctgaaactg atggcctttc accttctgtt
     2401 gcctctccaa gtcccaaaga agtcaatttt gtttcaaggg gagcttcaag tcaccagccc
     2461 agagttccac tttttcctga aaatggttta caccagcagc cagaaccctt gcttccaaat
     2521 aatatgaaat ctgcctgtga aaaacgttta ggatgttgta gttctcctca ttctaagcca
     2581 aattgctcaa ccctttctcc accaatgcca ctgccccagc tgttaccttc ggttactgat
     2641 gcaaggtcgg caggaccttc tgatcatatt gattcctcag ttactggggt tcaaaggttt
     2701 cgagatactc taaaaatacc ctacaagctg gaattaaaaa atgaaccagg gagaacggat
     2761 ttgaaacaca ttgttataga tgggagcaat gttgcaatta cccatggtct gaaaaagttc
     2821 ttttcttgtc gtggaattgc aattgcagtt gaatattttt ggaagcttgg caacagaaac
     2881 atcactgtat ttgtccctca gtggagaaca aggcgtgatc ctaatgtcac agaacagcac
     2941 ttcttaaccc agctccagga gctcggaata ttatctttaa ctcctgcccg gatggtcttt
     3001 ggagaaagaa ttgcttctca taatgacagg tttctactac acttagcgga caaaactggt
     3061 ggcataattg tgacaaatga taatttcaga gaatttgtga atgagtcagt ctcttggaga
     3121 gaaattatta caaaaaggct gctgcagtac acgttcgtgg gggacatatt tatggttccc
     3181 gatgatcctc tgggaagaag tggacctcga ttagaggagt ttcttcagaa ggaagtctgt
     3241 cttagagata tgcagcccct actcagtgcc ctgccaaatg tgggcatgtt tgaccccagc
     3301 ttcagagtcc ctggcaccca ggcagccagc accagccacc agcctccgac ccggattcag
     3361 ggagcccctt caagccactg gctccctcag cagccccact tcccactgct gccagccctc
     3421 cccagtctcc agcagaacct gcccatgcca gctcagagat cttctgcaga aaccaacgag
     3481 ctgagggaag cccttctgaa gatcttccct gactcagagc aaagactgaa aatagaccag
     3541 atcttggtgg cccacccata catgaaagat ctaaatgcgc tttctgccat ggtgttggat
     3601 ctcgagaccg gtgactacaa agaccatgac ggtgattata aagatcatga catcgattac
     3661 aaggatgacg atgacaagtg accggtaagt ttaaaccgct gatcagcctc gactgtgcct
     3721 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3781 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3841 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3901 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     3961 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4021 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     4081 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4141 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     4201 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4261 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4321 tcggtctatt cttttgattt ataagggatt ttggccattt cggcctattg gttaaaaaat
     4381 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     4441 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4501 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4561 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4621 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4681 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4741 taggcttttg caaaaagctc ccgggatgac cgagtacaag cccacggtgc gcctcgccac
     4801 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     4861 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     4921 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     4981 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     5041 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     5101 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     5161 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     5221 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     5281 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     5341 catgacccgc aagcccggtg cctgattcga aatgaccgac caagcgacgc ccaacctgcc
     5401 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     5461 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     5521 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     5581 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     5641 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     5701 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5761 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5821 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5881 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5941 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6001 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6061 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6121 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6181 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6241 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     6301 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6361 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6421 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6481 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     6541 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6601 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac
     6661 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6721 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6781 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6841 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6901 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6961 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7021 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7081 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7141 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7201 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7261 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7321 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7381 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7441 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7501 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7561 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7621 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7681 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7741 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7801 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
