LOCUS       V035459                 7651 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035459
VERSION     V035459
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             706..2310
                     /label="CCT2(NM_006431)"
                     /note="CCT2(NM_006431)"
                     /gene="CCT2"
     CDS             2317..2343
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2347..2373
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2377..2403
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    2427..3015
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3081..3129
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3331..3759
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3773..4102
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4169..4963
                     /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    5139..5260
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5309..5325)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5333..5349
                     /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(5357..5387)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5402..5423
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5711..6299)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6470..7330)
                     /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(7331..7435)
                     /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 ggatccacta gtccagtgtg gtggaattcg ccaccatggc gtccctttcc
      721 cttgcacctg ttaacatctt taaggcagga gctgatgaag agagagcaga gacagctcgt
      781 ctgacttctt ttattggtgc catcgccatt ggagacttgg taaagagcac cttgggaccc
      841 aaaggcatgg acaaaattct tctaagcagt ggacgagatg cctctcttat ggtaaccaat
      901 gatggtgcca ctattctaaa aaacattggt gttgacaatc cagcagctaa agttttagtt
      961 gatatgtcaa gggttcaaga tgatgaagtt ggtgatggca ctacctctgt taccgtttta
     1021 gcagcagaat tattaaggga agcagaatct ttaattgcaa aaaagattca tccacagacc
     1081 atcatagcgg gttggagaga agccacgaag gctgcaagag aggcgctgtt gagttctgca
     1141 gttgatcatg gttccgatga agttaaattc cgtcaagatt taatgaatat tgcgggcaca
     1201 acattatcct caaaacttct tactcatcac aaagaccact ttacaaagtt agctgtagaa
     1261 gcagttctca gactgaaagg ctctggcaac ctggaggcaa ttcatattat caagaagcta
     1321 ggaggaagtt tggcagattc ctatttagat gaaggcttcc tgttggataa aaaaattgga
     1381 gtaaatcaac caaaacgaat tgaaaatgct aaaattctta ttgcaaatac tggtatggat
     1441 acagacaaaa taaagatatt tggttcccgg gtaagagttg actctacagc aaaggttgca
     1501 gaaatagaac atgcggaaaa ggaaaaaatg aaggagaaag ttgaacgtat tcttaagcat
     1561 ggaataaatt gctttattaa caggcaatta atttataatt atcctgaaca gctctttggt
     1621 gctgctggtg tcatggctat tgagcatgca gattttgcag gtgtggaacg cctagctctt
     1681 gtcacaggtg gtgaaattgc ctctaccttt gatcacccag aactggtgaa gcttggaagt
     1741 tgcaaactta tcgaggaagt catgattgga gaagacaaac tcattcactt ttctggggtt
     1801 gcccttggtg aggcttgtac cattgttttg cgtggtgcca ctcaacaaat tttagatgaa
     1861 gcagaaagat cattgcatga tgctctttgt gttcttgcgc aaactgtaaa ggactctaga
     1921 acagtttatg gaggaggctg ttctgagatg ttgatggctc atgctgtgac acagcttgcc
     1981 aatagaacac caggcaaaga agctgttgca atggagtctt atgctaaagc actgagaatg
     2041 ttgccaacca tcatagctga caatgcaggc tatgacagtg cagacctggt ggcacagctc
     2101 agggctgctc acagtgaagg caataccact gctggattgg atatgaggga aggcaccatt
     2161 ggagatatgg ctatcctggg tataacagaa agttttcaag tgaagcgaca ggttcttctg
     2221 agtgcagctg aagcagcaga ggtgattctg cgtgtggaca acatcatcaa agcggcaccc
     2281 aggaaacgtg tccctgatca ccacccctgt ctcgagtacc catacgacgt acctgattat
     2341 gcaacttatc cttatgatgt acctgattat gctagctatc cttatgatgt cccagactac
     2401 gcatagtgag ggcccgatat ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat
     2461 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     2521 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     2581 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     2641 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     2701 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     2761 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     2821 agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     2881 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     2941 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     3001 gggccgcctc cccgcctgga aacgggggag gctaactgaa acacggaagg agacaatacc
     3061 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt
     3121 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     3181 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     3241 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcagatc
     3301 tgcgcagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     3361 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3421 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3481 atcggggcat ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3541 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3601 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3661 accctatctc ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt
     3721 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     3781 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3841 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3901 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3961 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4021 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     4081 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     4141 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     4201 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4261 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4321 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     4381 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4441 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4501 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4561 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4621 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4681 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4741 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4801 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4861 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4921 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4981 gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     5041 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5101 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     5161 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5221 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     5281 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5341 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5401 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5461 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5521 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5581 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5641 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5701 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5761 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5821 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5881 cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg
     5941 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6001 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6061 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6121 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     6181 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6241 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6301 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6361 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6421 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6481 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6541 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6601 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6661 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6721 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6781 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6841 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6901 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6961 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7021 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7081 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7141 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7201 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7261 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7321 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7381 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7441 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtc
     7501 gactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
     7561 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
     7621 gaccgacaat tgcatgaaga atctgcttag g
//
