LOCUS       V035027                 9686 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035027
VERSION     V035027
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             908..5125
                     /label="USP6(NM_004505)"
                     /note="USP6(NM_004505)"
                     /gene="USP6"
     CDS             5132..5158
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             5162..5188
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             5192..5218
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    5253..5477
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5523..5951
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5965..6294
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6361..7155
                     /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    7329..7450
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7499..7515)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7523..7539
                     /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(7547..7577)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7592..7613
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7901..8486)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8657..9517)
                     /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(9518..9622)
                     /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 ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg gacatggtag agaatgcaga tagtttgcag gcacaggagc ggaaggacat
      961 acttatgaag tatgacaagg gacaccgagc tgggctgcca gaggacaagg ggcctgagcc
     1021 cgttggaatc aacagcagca ttgatcgttt tggcattttg catgagacgg agctgcctcc
     1081 tgtgactgca cgggaggcga agaaaattcg gcgggagatg acacgaacga gcaagtggat
     1141 ggaaatgctg ggagaatggg agacatataa gcacagtagc aaactcatag atcgagtgta
     1201 caagggaatt cccatgaaca tccggggccc ggtgtggtca gtcctcctga acattcagga
     1261 aatcaagttg aaaaaccccg gaagatacca gatcatgaag gagaggggca agaggtcatc
     1321 tgaacacatc caccacatcg acctggacgt gaggacgact ctccggaacc atgtcttctt
     1381 tagggatcga tatggagcca agcagaggga actattctac atcctcctgg cctattcgga
     1441 gtataacccg gaggtgggct actgcaggga cctgagccac atcaccgcct tgttcctcct
     1501 ttatctgcct gaggaggacg cattctgggc actggtgcag ctgctggcca gtgagaggca
     1561 ctccctgcca ggattccaca gcccaaatgg tgggacagtc caggggctcc aagaccaaca
     1621 ggagcatgtg gtacccaagt cacaacccaa gaccatgtgg catcaggaca aggaaggtct
     1681 atgcgggcag tgtgcctcgt taggctgcct tctccggaac ctgattgacg ggatctctct
     1741 cgggctcacc ctgcgcctgt gggacgtgta tttggtggaa ggagaacagg tgttgatgcc
     1801 aataaccagc attgctctta aggttcagca gaagcgcctc atgaagacat ccaggtgtgg
     1861 cctgtgggca cgtctgcgga accaattctt cgatacctgg gccatgaacg atgacaccgt
     1921 gctcaagcat cttagggcct ctacgaagaa actaacaagg aagcaagggg acctgccacc
     1981 cccagccaaa cgcgagcaag ggtccttggc acccaggcct gtgccggctt cacgtggtgg
     2041 gaagaccctc tgcaaggggt ataggcaggc ccctccaggc ccaccagccc agttccagcg
     2101 gcccatttgc tcagcttccc cgccatgggc atctcgtttt tccacgccct gtcctggtgg
     2161 ggctgtccgg gaagacacgt accctgtggg cactcagggt gtgcccagcc tggccctggc
     2221 tcagggagga cctcagggtt cctggagatt cctggagtgg aagtcaatgc cccggctccc
     2281 aacggacctg gatatagggg gcccttggtt cccccattat gattttgaat ggagctgctg
     2341 ggtccgtgcc atatcccagg aggaccagct ggccacctgc tggcaggctg aacactgcgg
     2401 agaggttcac aacaaagata tgagttggcc tgaggagatg tcttttacag caaatagtag
     2461 taaaatagat agacaaaagg ttcccacaga aaagggagcc acaggtctaa gcaacctggg
     2521 aaacacatgc ttcatgaact caagcatcca gtgcgttagt aacacacagc cactgacaca
     2581 gtattttatc tcagggagac atctttatga actcaacagg acaaatccca ttggtatgaa
     2641 ggggcatatg gctaaatgct atggtgattt agtgcaggaa ctctggagtg gaactcagaa
     2701 gagtgttgcc ccattaaagc ttcggcggac catagcaaaa tatgctccca agtttgatgg
     2761 gtttcagcaa caagactccc aagaacttct ggcttttctc ttggatggtc ttcatgaaga
     2821 tctcaaccga gtccatgaaa agccatatgt ggaactgaag gacagtgatg gccgaccaga
     2881 ctgggaagta gctgcagagg cctgggacaa ccatctaaga agaaatagat caattattgt
     2941 ggatttgttc catgggcagc taagatctca agtcaaatgc aagacatgtg ggcatataag
     3001 tgtccgattt gaccctttca attttttgtc tttgccacta ccaatggaca gttacatgga
     3061 cttagaaata acagtgatta agttagatgg tactacccct gtacggtatg gactaagact
     3121 gaatatggat gaaaagtaca caggtttaaa aaaacagctg agggatctct gtggacttaa
     3181 ttcagaacaa atcctactag cagaagtaca tgattccaac ataaagaact ttcctcagga
     3241 taaccaaaaa gtacaactct cagtgagcgg atttttgtgt gcatttgaaa ttcctgtccc
     3301 ttcatctcca atttcagctt ctagtccaac acaaatagat ttctcctctt caccatctac
     3361 aaatggaatg ttcaccctaa ctaccaatgg ggacctaccc aaaccaatat tcatccccaa
     3421 tggaatgcca aacactgttg tgccatgtgg aactgagaag aacttcacaa atggaatggt
     3481 taatggtcac atgccatctc ttcctgacag cccctttaca ggttacatca ttgcagtcca
     3541 ccgaaaaatg atgaggacag aactgtattt cctgtcacct caggagaatc gccccagcct
     3601 ctttggaatg ccattgattg ttccatgcac tgtgcatacc cggaagaaag acctatatga
     3661 tgcggtttgg attcaagtat cctggttagc aagaccactc ccacctcagg aagctagtat
     3721 tcatgcccag gatcgtgata actgtatggg ctatcaatat ccattcactc tacgagttgt
     3781 gcagaaagat gggaactcct gtgcttggtg cccacagtat agattttgca gaggctgtaa
     3841 aattgattgt ggggaagaca gagctttcat tggaaatgcc tatattgctg tggattggca
     3901 ccccacagcc cttcaccttc gctatcaaac atcccaggaa agggttgtag ataagcatga
     3961 gagtgtggag cagagtcggc gagcgcaagc cgagcccatc aacctggaca gctgtctccg
     4021 tgctttcacc agtgaggaag agctagggga aagtgagatg tactactgtt ccaagtgtaa
     4081 gacccactgc ttagcaacaa agaagctgga tctctggagg cttccaccct tcctgattat
     4141 tcaccttaag cgatttcaat ttgtaaatga tcagtggata aaatcacaga aaattgtcag
     4201 atttcttcgg gaaagttttg atccgagtgc ttttttggta ccacgagacc cggccctctg
     4261 ccagcataaa ccactcacac cccaggggga tgagctctcc aagcccagga ttctggcaag
     4321 agaggtgaag aaagtggatg cgcagagttc ggctggaaaa gaggacatgc tcctaagcaa
     4381 aagcccatcc tcactcagcg ctaacatcag cagcagccca aaaggttctc cttcttcatc
     4441 aagaaaaagt ggaaccagct gtccctccag caaaaacagc agccctaata gcagcccacg
     4501 gactttgggg aggagcaaag ggaggctccg gctgccccag attggcagca aaaataagcc
     4561 gtcaagtagt aagaagaact tggatgccag caaagagaat ggggctgggc agatctgtga
     4621 gctggctgac gccttgagcc gagggcatat gcgggggggc agccaaccag agctggtcac
     4681 tcctcaggac catgaggtag ctttggccaa tggattcctt tatgagcatg aagcatgtgg
     4741 caatggctgt ggcgatggct acagcaatgg tcagcttgga aaccacagtg aagaagacag
     4801 cactgatgac caaagagaag acactcatat taagcctatt tataatctat atgcaatttc
     4861 atgccattca ggaattctga gtgggggcca ttacatcact tatgccaaaa acccaaactg
     4921 caagtggtac tgttataatg acagcagctg tgaggaactt caccctgatg aaattgacac
     4981 cgactctgcc tacattcttt tctatgagca gcaggggata gactacgcac aatttctgcc
     5041 aaagattgat ggcaaaaaga tggcagacac aagcagtacg gatgaagact ctgagtctga
     5101 ttacgaaaag tactctatgt tacagctcga gtacccatac gacgtacctg attatgcaac
     5161 ttatccttat gatgtacctg attatgctag ctatccttat gatgtcccag actacgcatg
     5221 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     5281 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     5341 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     5401 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     5461 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     5521 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     5581 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     5641 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     5701 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     5761 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     5821 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     5881 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     5941 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     6001 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     6061 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     6121 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     6181 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     6241 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     6301 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     6361 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     6421 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     6481 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     6541 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     6601 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     6661 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     6721 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     6781 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     6841 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     6901 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     6961 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     7021 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     7081 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     7141 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     7201 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     7261 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     7321 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     7381 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     7441 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     7501 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     7561 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     7621 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     7681 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     7741 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     7801 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     7861 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     7921 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     7981 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     8041 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     8101 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     8161 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     8221 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     8281 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     8341 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     8401 gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga
     8461 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     8521 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     8581 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     8641 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     8701 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     8761 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     8821 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     8881 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     8941 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     9001 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     9061 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     9121 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     9181 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     9241 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     9301 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     9361 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     9421 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     9481 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     9541 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     9601 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     9661 cgggagatct cccgatcccc tatggt
//
