LOCUS       V031828                 8872 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031828
VERSION     V031828
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..3531
                     /label="USP33(NM_015017)"
                     /note="USP33(NM_015017)"
                     /gene="USP33"
     CDS             3538..3564
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3568..3594
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3598..3624
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    3648..4236
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4302..4350
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4552..4980
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4994..5323
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5390..6184
                     /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    6360..6481
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6530..6546)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6554..6570
                     /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(6578..6608)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6623..6644
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6932..7520)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7691..8551)
                     /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(8552..8656)
                     /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 ccaccatgac aggatcaaat
      721 tcacacataa cgatattaac cttaaaggtg ttacctcatt ttgaaagtct tgggaaacag
      781 gaaaaaattc ctaacaaaat gtcagctttt cgaaatcatt gtccacattt ggattcagtt
      841 ggtgaaataa caaaagaaga tttgatacaa aaatcccttg gtacttgtca ggattgtaaa
      901 gtccaaggac caaatctttg ggcatgtctg gagaatagat gttcatatgt tggctgtggt
      961 gaatcacaag tagatcacag caccatacat tctcaggaga caaagcatta tctaactgtg
     1021 aaccttacca ctcttcgagt atggtgttat gcttgcagca aagaagtatt tttggatagg
     1081 aaattaggaa ctcagccttc attgcctcat gtaagacaac ctcaccaaat acaagaaaac
     1141 agtgtccagg attttaaaat acccagtaat acaacattaa aaactcctct ggttgccgta
     1201 tttgatgatc tggatataga agcggatgaa gaagatgaac ttagggccag aggtcttaca
     1261 ggtttgaaaa atattggaaa tacttgttac atgaatgcag ctttgcaggc tctttctaat
     1321 tgcccacctt tgacacagtt ttttcttgat tgtggaggac tagctcgaac agataagaaa
     1381 cctgccattt gtaaaagtta tctcaaacta atgacagagc tgtggcataa aagcaggcca
     1441 ggatctgttg tgcctactac tctgtttcaa ggaattaaaa ctgtaaatcc aacatttcgg
     1501 gggtattctc agcaggatgc tcaagaattc cttcgatgtt taatggattt gcttcatgaa
     1561 gaattgaaag agcaagtcat ggaagtagaa gaagatccgc aaaccataac cactgaggag
     1621 acaatggaag aagacaagag ccagtcggat gtagattttc agtcttgtga atcttgtagc
     1681 aacagtgata gagcagaaaa tgaaaatggc tctagatgct tttctgaaga taataatgaa
     1741 acaacaatgt taattcagga tgatgaaaac aattcagaaa tgtcaaagga ttggcaaaaa
     1801 gagaagatgt gcaataagat taataaagta aattctgaag gcgaatttga taaagataga
     1861 gactctatat ctgaaacagt cgacttaaac aaccaggaaa ctgtcaaagt gcaaatacac
     1921 agcagagctt cagaatatat cactgatgtc cattcgaatg acctgtctac accacagatc
     1981 cttccatcaa atgaaggtgt taatccacgt ttatcggcaa gccctcctaa atcaggcaat
     2041 ttgtggccag gattggcacc accacacaaa aaagctcagt ctgcatctcc aaagagaaaa
     2101 aaacagcaca agaaatacag aagtgttatt tcagacatat ttgatggaac aatcattagt
     2161 tcagtgcagt gtctgacttg tgacagggtg tctgtaaccc tcgagacctt tcaagatctg
     2221 tccttgccaa ttcctggcaa ggaagacctt gctaagctgc attcatcaag tcatccaact
     2281 tctatagtca aagcaggatc atgtggcgaa gcatatgctc cacaagggtg gatagctttt
     2341 ttcatggaat atgtgaagag gtttgttgtc tcatgtgtcc ctagctggtt ttggggtcca
     2401 gtagtaacct tgcaagattg tcttgctgcc ttctttgcca gagatgaact aaaaggtgac
     2461 aatatgtaca gttgtgaaaa atgcaaaaag ttgagaaatg gagtgaagtt ttgtaaagta
     2521 caaaactttc ctgagatttt gtgcatccac cttaaaagat tcagacatga actaatgttt
     2581 tccaccaaaa tcagtaccca tgtttcattt ccgctagaag gcttggatct tcagccattt
     2641 cttgctaagg atagtccagc tcaaattgtg acatatgatc ttctgtcagt catttgccat
     2701 catggaactg caagtagtgg acactatata gcctactgcc gaaacaatct aaataatctc
     2761 tggtatgaat ttgatgatca gagtgtcact gaagtttcag aatctactgt acaaaatgca
     2821 gaagcttacg ttcttttcta taggaagagc agcgaagagg cacaaaaaga gaggagaagg
     2881 atatcaaatt tattgaacat aatggaacca agcctccttc agttttatat ttctcgacag
     2941 tggcttaata aatttaagac ctttgccgaa cctggcccta tttcaaataa tgactttctt
     3001 tgtattcatg gaggtgttcc tccaagaaaa gctggttata ttgaagacct ggttttgatg
     3061 ctgcctcaga acatttggga taacctatat agcaggtatg gtggaggacc agctgtcaac
     3121 catctgtaca tttgtcatac ttgccaaatt gaggcggaga aaattgaaaa aagaagaaaa
     3181 actgaattgg aaatttttat tcggcttaac agagcgttcc aaaaagagga ctctccagct
     3241 actttttatt gcatcagtat gcagtggttt agagaatggg aaagttttgt gaagggtaaa
     3301 gatggagatc ctccaggtcc tattgacaat actaagattg cagtcactaa atgtggtaat
     3361 gtgatgctta ggcaaggagc agattctggc cagatttctg aagaaacatg gaattttctg
     3421 cagtctattt atggtggagg gcctgaagtt atcctgcgac ctccggttgt tcatgttgat
     3481 ccagatatac ttcaagcaga agaaaaaatt gaagtagaaa ctcggtcttt gctcgagtac
     3541 ccatacgacg tacctgatta tgcaacttat ccttatgatg tacctgatta tgctagctat
     3601 ccttatgatg tcccagacta cgcatagtga gggcccgata tctcgacaat caacctctgg
     3661 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     3721 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     3781 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     3841 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     3901 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     3961 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     4021 attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca
     4081 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     4141 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     4201 agacgagtcg gatctccctt tgggccgcct ccccgcctgg aaacggggga ggctaactga
     4261 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     4321 taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     4381 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     4441 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     4501 caggccctgc catagcagat ctgcgcagct ggggctctag ggggtatccc cacgcgccct
     4561 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4621 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4681 gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac
     4741 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     4801 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4861 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     4921 tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4981 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     5041 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     5101 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     5161 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     5221 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     5281 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     5341 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     5401 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     5461 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     5521 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     5581 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     5641 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     5701 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     5761 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     5821 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     5881 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     5941 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     6001 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     6061 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     6121 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     6181 ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     6241 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     6301 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     6361 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     6421 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     6481 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     6541 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     6601 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     6661 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     6721 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     6781 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     6841 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     6901 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6961 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     7021 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     7081 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta
     7141 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     7201 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     7261 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     7321 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     7381 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7441 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     7501 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     7561 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7621 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7681 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7741 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7801 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7861 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     7921 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7981 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     8041 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     8101 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     8161 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     8221 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     8281 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     8341 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     8401 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     8461 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     8521 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     8581 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8641 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     8701 atctcccgat cccctatggt cgactctcag tacaatctgc tctgatgccg catagttaag
     8761 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
     8821 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gg
//
