LOCUS       V047630                 8328 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047630
VERSION     V047630
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             875..895
                     /codon_start=1
                     /locus_tag=""
                     /note=""
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /protein_id=""
                     /transl_table=1
                     /translation="PKKKRKV"
                     /label="SV40 NLS"
     CDS             902..3223
                     /label="RFWD3(NM_018124)"
                     /note="RFWD3(NM_018124)"
                     /gene="RFWD3"
     CDS             3224..3289
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             3320..4039
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK*"
                     /label="EGFP"
     polyA_signal    4090..4314
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4360..4788
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4802..5131
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5198..5797
                     /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    5971..6092
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6141..6157)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6165..6181
                     /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(6189..6219)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6234..6255
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6543..7128)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7299..8159)
                     /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(8160..8264)
                     /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 ggctagccac catgccaaag aagaagcgga aggtcggttc
      901 aatggctcat gaagcaatgg aatatgatgt tcaggtgcag ttaaatcatg ccgaacaaca
      961 gccagctcct gctggcatgg ccagcagcca agggggacca gccctcctcc agcctgttcc
     1021 tgctgatgtg gtcagcagcc agggggtacc atccatcctc cagccagctc ctgctgaggt
     1081 gatcagcagc caagcgacac cacccctgct ccagcctgct ccgcaactgt ctgttgacct
     1141 gacagaagtg gaggtcttgg gagaagacac tgtggagaac atcaatccaa gaacttcaga
     1201 acaacatagg cagggatctg atggtaatca caccatccca gcatcttcgt tgcattcaat
     1261 gaccaacttc atcagcggac tgcagagact tcatggcatg ctggaattcc tgagaccttc
     1321 atcttcaaac cacagtgtag ggccaatgag aacaagaagg agggtatctg cttcacggag
     1381 ggcaagagcc ggagggtctc agaggacaga cagtgccagg ttgagagcac cattggatgc
     1441 ttactttcag gtgagcagga cccagcctga cttgccagct accacttatg attcagagac
     1501 taggaatcct gtatctgaag agttgcaggt gtctagtagt tctgattctg acagtgacag
     1561 ctctgcagag tatggagggg ttgttgacca ggcagaggaa tctggagctg tcattttaga
     1621 agagcaacta gcaggtgtct cagcagagca agaagttaca tgtatcgatg gaggcaagac
     1681 cctccccaaa cagccatctc cccagaagtc tgagcctctg ctaccttctg cttctatgga
     1741 tgaggaagaa ggggacactt gtacaatatg tctggaacag tggaccaatg ctggggacca
     1801 ccggctctca gcattacgct gtgggcatct ctttgggtat aggtgcattt ccacgtggct
     1861 taaaggacaa gtacgaaaat gtccccagtg caacaagaaa gccaggcaca gtgacattgt
     1921 cgtcctttat gcccgaaccc tgagagcttt ggacactagt gaacaggagc gcatgaaaag
     1981 ttccctactg aaggaacaga tgctaaggaa acaggccgag ttagaatcag cacagtgccg
     2041 actccaactg caggtcctca ctgataagtg cactaggctt caaaggcgtg ttcaggactt
     2101 gcaaaaactt acgtcacatc aaagtcagaa tttacagcaa cccaggggct cccaagcatg
     2161 ggtcctgagc tgctcaccct ccagccaggg ccagcacaag cacaagtacc acttccaaaa
     2221 gaccttcaca gtatctcagg caggaaactg ccggatcatg gcatactgtg atgctctgag
     2281 ctgcctggtg atatcacagc cttctcctca ggcctctttt cttccaggct ttggtgttaa
     2341 gatgttgagt actgccaaca tgaagagcag tcagtacatt ccgatgcatg gcaaacagat
     2401 ccgtggactg gcgtttagca gttacctcag aggcttgcta ctctctgctt ccctagacaa
     2461 cactattaaa ctgaccagcc tggagacaaa taccgtggtc cagacttata atgctggacg
     2521 tcctgtctgg agctgttgct ggtgtcttga tgaggctaac tacatctatg ctggactggc
     2581 caatggttca attctggtat atgacgtgcg aaacacgagc agtcatgtgc aggagttagt
     2641 agctcagaaa gccagatgcc cactggtctc cctgtcatac atgcccagag ctgcctcagc
     2701 tgcatttcca tatggtgggg tgctggctgg aaccttggag gatgcttcat tctgggaaca
     2761 gaaaatggac ttttctcatt ggcctcatgt gctgcccttg gagccagggg gctgcataga
     2821 ctttcagaca gagaacagct cccggcactg tcttgtgacc tacaggcctg ataaaaatca
     2881 caccaccata cgaagtgtgc tgatggaaat gtcctaccga ctggatgaca ctggaaatcc
     2941 aatctgctcc tgccagcctg tacatacatt ttttggagga cctacttgca aactattgac
     3001 caaaaatgcc attttccaaa gcccagagaa tgatggcaac atcctggtgt gtactgggga
     3061 tgaagcagca aattctgccc tgctgtggga tgctgccagt ggctcgttgc tccaggacct
     3121 acagaccgat cagcctgtgt tggacatctg cccatttgag gtgaaccgta acagctactt
     3181 ggctacctta acagagaaga tggtccacat ctataagtgg gaggactaca aagaccatga
     3241 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagg gtggaggcgg
     3301 atcaggaggc ggtggatcta tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     3361 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     3421 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     3481 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     3541 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     3601 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     3661 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     3721 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     3781 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     3841 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     3901 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     3961 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     4021 ggacgagctg tacaagtaag cggccgctcg agtctagagg gcccgtttaa acccgctgat
     4081 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     4141 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     4201 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     4261 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     4321 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     4381 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4441 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4501 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4561 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4621 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4681 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4741 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     4801 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4861 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4921 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4981 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     5041 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     5101 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     5161 cggatctgat caagagacag gatgaggatc gtttcgcatg accgagtaca agcccacggt
     5221 gcgcctcgcc acccgcgacg acgtcccccg ggccgtacgc accctcgccg ccgcgttcgc
     5281 cgactacccc gccacgcgcc acaccgtcga cccggaccgc cacatcgagc gggtcaccga
     5341 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     5401 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     5461 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     5521 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     5581 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     5641 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
     5701 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     5761 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgagcg ggactctggg gttcgaaatg
     5821 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5881 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5941 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     6001 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     6061 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     6121 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     6181 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     6241 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     6301 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     6361 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     6421 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     6481 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     6541 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6601 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6661 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6721 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6781 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6841 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6901 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6961 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     7021 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     7081 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     7141 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     7201 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     7261 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     7321 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     7381 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     7441 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     7501 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     7561 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     7621 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     7681 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     7741 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     7801 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     7861 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     7921 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     7981 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     8041 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     8101 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     8161 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     8221 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     8281 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggt
//
