LOCUS       V022718                 8000 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022718
VERSION     V022718
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"
     CDS             677..2680
                     /label="CD6(NM_006725)"
                     /note="CD6(NM_006725)"
                     /gene="CD6"
     CDS             2687..2752
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2776..3364
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3430..3478
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3680..4108
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4122..4451
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4518..5312
                     /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    5488..5609
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5658..5674)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5682..5698
                     /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(5706..5736)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5751..5772
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6060..6648)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6819..7679)
                     /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(7680..7784)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc gccaccatgt ggctcttctt cgggatcact ggattgctga cggcagccct
      721 ctcaggtcat ccatctccag ccccacctga ccagctcaac accagcagtg cagagagtga
      781 gctctgggag ccaggggagc ggcttccggt ccgtctgaca aacgggagca gcagctgcag
      841 cgggacggtg gaggtgcggc tcgaggcgtc ctgggagccc gcgtgcgggg cgctctggga
      901 cagccgcgcc gccgaggccg tgtgccgagc actgggctgc ggcggggcgg aggccgcctc
      961 tcagctcgcc ccgccgaccc ctgagctgcc gcccccgcct gcagccggga acaccagcgt
     1021 agcagctaat gccactctgg ccggggcgcc cgccctcctg tgcagcggcg ccgagtggcg
     1081 gctctgcgag gtggtggagc acgcgtgccg cagcgacggg aggcgggccc gtgtcacctg
     1141 tgcagagaac cgcgcgctgc gcctggtgga cggtggcggc gcctgcgccg gccgcgtgga
     1201 gatgctggag catggcgagt ggggatcagt gtgcgatgac acttgggacc tggaggacgc
     1261 ccacgtggtg tgcaggcaac tgggctgcgg ctgggcagtc caggccctgc ccggcttgca
     1321 cttcacgccc ggccgcgggc ctatccaccg ggaccaggtg aactgctcgg gggccgaagc
     1381 ttacctgtgg gactgcccgg ggctgccagg acagcactac tgcggccaca aagaggacgc
     1441 gggcgcggtg tgctcagagc accagtcctg gcgcctgaca gggggcgctg accgctgcga
     1501 ggggcaggtg gaggtacact tccgaggggt ctggaacaca gtgtgtgaca gtgagtggta
     1561 cccatcggag gccaaggtgc tctgccagtc cttgggctgt ggaactgcgg ttgagaggcc
     1621 caaggggctg ccccactcct tgtccggcag gatgtactac tcatgcaatg gggaggagct
     1681 caccctctcc aactgctcct ggcggttcaa caactccaac ctctgcagcc agtcgctggc
     1741 agccagggtc ctctgctcag cttcccggag tttgcacaat ctgtccactc ccgaagtccc
     1801 tgcaagtgtt cagacagtca ctatagaatc ttctgtgaca gtgaaaatag agaacaagga
     1861 atctcgggag ctaatgctcc tcatcccctc catcgttctg ggaattctcc tccttggctc
     1921 cctcatcttc atagccttca tcctcttgag aattaaagga aaatatgccc tccccgtaat
     1981 ggtgaaccac cagcacctac ccaccaccat cccggcaggg agcaatagct atcaaccggt
     2041 ccccatcacc atccccaaag aagttttcat gctgcccatc caggtccagg ccccgccccc
     2101 tgaggactca gactctggct cggactcaga ctatgagcac tatgacttca gcgcccagcc
     2161 tcctgtggcc ctgaccacct tctacaattc ccagcggcat cgggtcacag atgaggaggt
     2221 ccagcaaagc aggttccaga tgccaccctt ggaggaagga cttgaagagt tgcatgcctc
     2281 ccacatccca actgccaacc ctggacactg cattacagac ccgccatccc tgggccctca
     2341 gtatcacccg aggagcaaca gtgagtcgag cacctcttca ggggaggatt actgcaatag
     2401 tcccaaaagc aagctgcctc catggaaccc ccaggtgttt tcttcagaga ggagttcctt
     2461 cctggagcag cccccaaact tggagctggc cggcacccag ccagcctttt cagcagggcc
     2521 cccggctgat gacagctcca gcacctcatc cggggagtgg taccagaact tccagccacc
     2581 accccagccc ccttcggagg agcagtttgg ctgtccaggg tcccccagcc ctcagcctga
     2641 ctccaccgac aacgatgact acgatgacat cagcgcagcc ctcgaggact acaaagacca
     2701 tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca agtagtgagg
     2761 gcccgatatc tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     2821 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     2881 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     2941 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     3001 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     3061 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     3121 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc
     3181 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     3241 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     3301 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     3361 ccgcctggaa acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     3421 gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat
     3481 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     3541 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     3601 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcagatct gcgcagctgg
     3661 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3721 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3781 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc
     3841 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3901 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3961 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     4021 gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag
     4081 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     4141 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4201 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4261 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4321 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4381 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4441 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     4501 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4561 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4621 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4681 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4741 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4801 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4861 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4921 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4981 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     5041 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     5101 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     5161 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     5221 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     5281 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc
     5341 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     5401 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     5461 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     5521 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     5581 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     5641 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5701 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     5761 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5821 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5881 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5941 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     6001 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     6061 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6121 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     6181 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     6241 gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     6301 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6361 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6421 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6481 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6541 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6601 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6661 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6721 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6781 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6841 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6901 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6961 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     7021 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     7081 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     7141 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     7201 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     7261 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     7321 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     7381 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     7441 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     7501 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     7561 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     7621 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     7681 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     7741 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     7801 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtcg actctcagta
     7861 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     7921 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     7981 gcatgaagaa tctgcttagg
//
