LOCUS       V037204                 7649 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037204
VERSION     V037204
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             896..3247
                     /label="RBSN(NM_022340)"
                     /note="RBSN(NM_022340)"
                     /gene="RBSN"
     CDS             3278..3301
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin®, an engineered
                     form of streptavidin"
                     /transl_table=1
                     /translation="WSHPQFEK"
                     /label="Strep-Tag II"
     CDS             3341..3364
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin®, an engineered
                     form of streptavidin"
                     /transl_table=1
                     /translation="WSHPQFEK"
                     /label="Strep-Tag II"
     polyA_signal    3411..3635
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3681..4109
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4123..4452
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4519..5118
                     /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    5292..5413
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5462..5478)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5486..5502
                     /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(5510..5540)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5555..5576
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5864..6449)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6620..7480)
                     /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(7481..7585)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg ccaccatggc
      901 ttctctggac gacccagggg aagtgaggga gggcttcctc tgccctctgt gcctgaagga
      961 tctgcagtct ttctatcagc ttcactcaca ttacgaggaa gaacactcag gggaagaccg
     1021 tgatgtcaaa gggcaaatta aaagtcttgt ccagaaggct aaaaaagcaa aggacaggtt
     1081 gttgaaacga gaaggggatg atcgagcaga gtcagggacc caaggatatg agtctttcag
     1141 ctatggaggg gttgatcctt acatgtggga accccaggag cttggtgctg tgagaagcca
     1201 tctttccgac ttcaaaaaac accgagctgc tagaattgac cactatgttg tggaagtcaa
     1261 taaactaata atcaggttag agaagctcac tgcatttgac agaacaaata ctgagtctgc
     1321 aaagattcga gcaatagaaa agtctgtggt gccttgggtc aacgaccagg atgtcccttt
     1381 ctgtccagac tgtgggaata agttcagcat ccggaaccgc cgccaccact gccgcctctg
     1441 cgggtctatt atgtgcaaga agtgtatgga gctcatcagc cttcccttgg caaacaagct
     1501 caccagtgcc agcaaggagt ccctgagcac ccacaccagc cccagccagt cacccaacag
     1561 tgtccatggc tcccgccgag gcagcatcag cagcatgagc agtgtcagct cggtcctgga
     1621 tgagaaggac gatgaccgga tccgctgctg tacacactgc aaggacacgc tgctcaagag
     1681 agagcagcag attgatgaga aggagcacac acctgacatc gtgaagctct acgagaaatt
     1741 acgactttgc atggagaaag ttgaccagaa agctccagaa tacatcagga tggcagcatc
     1801 attaaatgct ggggagacaa cctacagtct ggaacatgcc agtgaccttc gagtggaagt
     1861 gcagaaagtg tatgagctga tagacgcttt aagtaagaag atcttaacct tgggcttgaa
     1921 ccaggaccct ccaccacatc caagcaattt gcggctgcag agaatgatca gatactcagc
     1981 tacacttttt gtgcaggaaa agttgcttgg tttgatgtca ctgccaacca aagaacagtt
     2041 tgaggaactg aaaaagaaaa ggaaggagga aatggagagg aagagggccg tggagagaca
     2101 agctgccctg gagtcccagc gaaggcttga ggaaaggcag agtggcctgg cttctcgagc
     2161 ggccaacggg gaggtggcat ctctccgcag gggccctgcc cccttgagaa aggctgaggg
     2221 ctggctccca ctgtcaggag gtcaggggca gagtgaggac tcagacccgc tcctccagca
     2281 gatccacaac atcacatcat tcatcaggca ggccaaggcc gcgggccgca tggatgaagt
     2341 gcgcactctg caggagaacc tgcggcagct gcaggacgag tatgaccagc agcagacaga
     2401 gaaggccatc gagctgtccc ggaggcaggc tgaggaggag gacctgcagc gggaacagct
     2461 gcagatgttg cgtgaacggg agttggaacg agaaagggag cagtttcggg tggcatccct
     2521 gcacacacgg actcggtccc tggacttcag agaaatcggc ccttttcagc tggagcccag
     2581 cagagagcct cgcacccacc ttgcttatgc tttggatcta ggctcttccc cagttccaag
     2641 cagcacagct cccaagaccc cttcacttag ctcaactcaa cccaccagag tgtggtctgg
     2701 gcccccagcc gttggccagg agcgcttacc ccagagcagc atgccacagc aacatgaggg
     2761 gccctcctta aacccctttg atgaggaaga cctctccagc cccatggaag aggccactac
     2821 tggtcctcct gctgcagggg tttccttaga cccttcagcc cgcatcctga aagagtacaa
     2881 tcctttcgag gaagaggacg aggaggagga agcagtggca gggaatccat tcattcagcc
     2941 agacagccca gctcctaacc ccttcagtga ggaagacgaa catccccagc agaggctctc
     3001 aagccctctg gttcctggta acccctttga ggaacccacc tgtatcaacc cctttgagat
     3061 ggacagtgac agtgggccag aggctgagga gcccatagag gaagagctcc tcctgcagca
     3121 gatcgataac atcaaggcat acatctttga tgccaagcag tgcggccgcc tggatgaggt
     3181 agaggtgctg acagagaatc tgcgggagct gaagcacacc ctggccaagc agaagggggg
     3241 cactgacgaa ttcagtgata tcgcggccgc gggtagctgg agccatccgc agtttgaaaa
     3301 aggtggtggt agcggtggtg gttcaggtgg tagcagtgca tggtcacacc ctcagtttga
     3361 gaaatgactc gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     3421 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3481 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3541 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3601 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     3661 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3721 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3781 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3841 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3901 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3961 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     4021 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     4081 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     4141 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4201 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4261 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4321 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4381 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4441 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     4501 ggatgaggat cgtttcgcat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     4561 gacgtccccc gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     4621 cacaccgtcg acccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     4681 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     4741 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     4801 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg
     4861 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
     4921 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc
     4981 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     5041 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     5101 cgcaagcccg gtgcctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     5161 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5221 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5281 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5341 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5401 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     5461 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5521 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5581 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5641 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5701 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5761 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5821 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5881 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5941 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6001 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6061 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6121 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6181 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6241 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6301 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6361 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc
     6421 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6481 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6541 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6601 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6661 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6721 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6781 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6841 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6901 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6961 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7021 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7081 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7141 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7201 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7261 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7321 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7381 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7441 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7501 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7561 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     7621 gatcgggaga tctcccgatc ccctatggt
//
