LOCUS       V024580                 7740 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024580
VERSION     V024580
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2420
                     /label="PRF1(NM_001083116)"
                     /note="PRF1(NM_001083116)"
                     /gene="PRF1"
     CDS             2427..2492
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2516..3104
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3170..3218
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3420..3848
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3862..4191
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4258..5052
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5228..5349
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5398..5414)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5422..5438
                     /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(5446..5476)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5491..5512
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5800..6388)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6559..7419)
                     /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(7420..7524)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggc agcccgtctg ctcctcctgg
      781 gcatccttct cctgctgctg cccctgcccg tccctgcccc gtgccacaca gccgcacgct
      841 cagagtgcaa gcgcagccac aagttcgtgc ctggtgcatg gctggccggg gagggtgtgg
      901 acgtgaccag cctccgccgc tcgggctcct tcccagtgga cacacaaagg ttcctgcggc
      961 ccgacggcac ctgcaccctc tgtgaaaatg ccctacagga gggcaccctc cagcgcctgc
     1021 ctctggcgct caccaactgg cgggcccagg gctctggctg ccagcgccat gtaaccaggg
     1081 ccaaagtcag ctccactgaa gctgtggccc gggatgcggc tcgtagcatc cgcaacgact
     1141 ggaaggtcgg gctggacgtg actcctaagc ccaccagcaa tgtgcatgtg tctgtggccg
     1201 gctcacactc acaggcagcc aactttgcag cccagaagac ccaccaggac cagtacagct
     1261 tcagcactga cacggtggag tgccgcttct acagtttcca tgtggtacac actcccccgc
     1321 tgcaccctga cttcaagagg gccctcgggg acctgcccca ccacttcaac gcctccaccc
     1381 agcccgccta cctcaggctt atctccaact acggcaccca cttcatccgg gctgtggagc
     1441 tgggtggccg catatcggcc ctcactgccc tgcgcacctg cgagctggcc ctggaagggc
     1501 tcacggacaa cgaggtggag gactgcctga ctgtcgaggc ccaggtcaac ataggcatcc
     1561 acggcagcat ctctgccgaa gccaaggcct gtgaggagaa gaagaagaag cacaagatga
     1621 cggcctcctt ccaccaaacc taccgggagc gccactcgga agtggttggc ggccatcaca
     1681 cctccattaa cgacctgctg ttcgggatcc aggccgggcc cgagcagtac tcagcctggg
     1741 taaactcgct gcccggcagc cctggcctgg tggactacac cctggaaccc ctgcacgtgc
     1801 tgctggacag ccaggacccg cggcgggagg cactgaggag ggccctgagt cagtacctga
     1861 cggacagggc tcgctggagg gactgcagcc ggccgtgccc accagggcgg cagaagagcc
     1921 cccgagaccc atgccagtgt gtgtgccatg gctcagcggt caccacccag gactgctgcc
     1981 ctcggcagag gggcctggcc cagctggagg tgaccttcat ccaagcatgg ggcctgtggg
     2041 gggactggtt cactgccacg gatgcctatg tgaagctctt ctttggtggc caggagctga
     2101 ggacgagcac cgtgtgggac aataacaacc ccatctggtc agtgcggctg gattttgggg
     2161 atgtgctcct ggccacaggg gggcccctga ggttgcaggt ctgggatcag gactctggca
     2221 gggacgatga cctccttggc acctgtgatc aggctcccaa gtctggttcc catgaggtga
     2281 gatgcaacct gaatcatggc cacctaaaat tccgctatca tgccaggtgc ttgccccacc
     2341 tgggaggagg cacctgcctg gactatgtcc cccaaatgct tctgggggag cctccaggaa
     2401 accggagtgg ggccgtgtgg ggatccgact acaaagacca tgacggtgat tataaagatc
     2461 atgacatcga ctacaaggat gacgatgaca agtagtgagg gcccgatatc tcgacaatca
     2521 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     2581 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     2641 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     2701 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     2761 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     2821 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     2881 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     2941 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     3001 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     3061 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg
     3121 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     3181 agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     3241 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     3301 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     3361 cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca
     3421 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3481 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3541 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag
     3601 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3661 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3721 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3781 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3841 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3901 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3961 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4021 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4081 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     4141 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4201 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4261 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4321 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4381 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     4441 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4501 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4561 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4621 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4681 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4741 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     4801 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4861 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4921 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4981 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5041 gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct
     5101 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5161 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5221 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5281 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5341 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5401 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5461 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5521 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5581 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5641 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5701 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5761 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5821 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5881 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5941 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     6001 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6061 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6121 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6181 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6241 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6301 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6361 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6421 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6481 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6541 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6601 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6661 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6721 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6781 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6841 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6901 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6961 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7021 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7081 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7141 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7201 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7261 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7321 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7381 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7441 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7501 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     7561 atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca
     7621 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     7681 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
