LOCUS       V026063                 7065 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026063
VERSION     V026063
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             898..924
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             940..2553
                     /label="KPNA1(NM_002264)"
                     /note="KPNA1(NM_002264)"
                     /gene="KPNA1"
     promoter        complement(2617..2635)
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /label="SP6 promoter"
     polyA_signal    2661..2885
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2931..3359
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3373..3703
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3770..4564
                     /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    4738..4859
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4908..4924)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4932..4948
                     /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(4956..4986)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5001..5022
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5310..5898)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6069..6929)
                     /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(6930..7034)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 ggagacccgc caccatgtac
      901 ccatacgacg tcccagacta cgctggaagc ttggtaccga tgaccacccc aggaaaagag
      961 aactttcgcc tgaaaagtta caagaacaaa tctctgaatc ccgatgagat gcgcaggagg
     1021 agggaggaag aaggactgca gttacgaaag cagaaaagag aagagcagtt attcaagcgg
     1081 agaaatgttg ctacagcaga agaagaaaca gaagaagaag ttatgtcaga tggaggcttt
     1141 catgaggctc agattagtaa catggagatg gcaccaggtg gtgtcatcac ttctgacatg
     1201 attgaaatga tattttccaa aagcccagag caacagcttt cagcaacaca gaaattcagg
     1261 aagctgcttt caaaagaacc taaccctcct attgatgaag ttatcagcac accaggagta
     1321 gtggccaggt ttgtggagtt cctcaaacga aaagagaatt gtacactgca gtttgaatca
     1381 gcttgggtac tgacaaatat tgcttcagga aattctcttc agacccgaat tgtgattcag
     1441 gcaggagctg tgcccatctt catagagttg ctcagctcag agtttgaaga tgtccaggaa
     1501 caggcagtct gggctcttgg caacattgct ggagatagta ccatgtgcag ggactatgtc
     1561 ttagactgca atatccttcc ccctcttttg cagttatttt caaagcaaaa ccgcctgacc
     1621 atgacccgga atgcagtatg ggctttgtct aatctctgta gagggaaaag tccacctcca
     1681 gaatttgcaa aggtttctcc atgtctgaat gtgctttcct ggttgctgtt tgtcagtgac
     1741 actgatgtac tggctgatgc ctgctgggcc ctctcatatc tatcagatgg acccaatgat
     1801 aaaattcaag cggtcatcga tgcgggagta tgtaggagac ttgtggaact gctgatgcat
     1861 aatgattata aagtggtttc tcctgctttg cgagctgtgg gaaacattgt cacaggggat
     1921 gatattcaga cacaggtaat tctgaattgc tcagctctgc agagtttatt gcatttgctg
     1981 agtagcccaa aggaatctat caaaaaggaa gcatgttgga cgatatctaa tattacagct
     2041 ggaaataggg cacagatcca gactgtgata gatgccaaca ttttcccagc cctcattagt
     2101 attttacaaa ctgctgaatt tcggacaaga aaagaagcag cttgggccat cacaaatgca
     2161 acttctggag gatcagctga acagatcaag tacctagtag aactgggttg tatcaagccg
     2221 ctctgtgatc tcctcacggt catggactct aagattgtac aggttgccct aaatggcttg
     2281 gaaaatatcc tgaggcttgg agaacaggaa gccaaaagga atggcactgg cattaaccct
     2341 tactgtgctt tgattgaaga agcttatggt ctggataaaa ttgagttctt acagagtcat
     2401 gaaaaccagg agatctacca aaaggccttt gatcttattg agcattactt cgggaccgaa
     2461 gatgaagaca gcagcattgc accccaggtt gaccttaacc agcagcagta catcttccaa
     2521 cagtgtgagg ctcctatgga aggtttccag ctttgagaat tctgcagata tccatcacac
     2581 tggcggccgc tcgagcatgc atctagaggg ccctattcta tagtgtcacc taaatgctag
     2641 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     2701 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     2761 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     2821 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     2881 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     2941 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3001 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3061 ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg
     3121 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3181 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3241 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3301 ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3361 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc aggcaggcag
     3421 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3481 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3541 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3601 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     3661 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     3721 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     3781 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     3841 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     3901 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     3961 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     4021 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     4081 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     4141 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     4201 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     4261 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     4321 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4381 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4441 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     4501 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     4561 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     4621 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     4681 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     4741 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     4801 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     4861 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     4921 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     4981 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     5041 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5101 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5161 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5221 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5281 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5341 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5401 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5461 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     5521 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5581 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5641 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5701 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     5761 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5821 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5881 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5941 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6001 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6061 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6121 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6181 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6241 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6301 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6361 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6421 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6481 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6541 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6601 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6661 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6721 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6781 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6841 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6901 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6961 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7021 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
