LOCUS       V041178                 7650 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041178
VERSION     V041178
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2330
                     /label="USP22(NM_015276)"
                     /note="USP22(NM_015276)"
                     /gene="USP22"
     CDS             2337..2402
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2426..3014
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3080..3128
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3330..3758
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3772..4101
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4168..4962
                     /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    5138..5259
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5308..5324)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5332..5348
                     /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(5356..5386)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5401..5422
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5710..6298)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6469..7329)
                     /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(7330..7434)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggt gtcccggcca gagcccgagg
      781 gcgaggccat ggacgccgag ctggcggtag cgccgccggg ctgctcgcac ctgggcagct
      841 tcaaggtgga caactggaag cagaacctgc gggccatcta ccagtgcttc gtgtggagcg
      901 gcacggctga ggcccgcaag cgcaaggcca agtcctgtat ctgccatgtc tgtggcgtcc
      961 acctcaacag gctgcattcc tgcctctact gtgtcttctt cggctgtttc acaaagaagc
     1021 atattcacga gcatgcgaag gcgaagcggc acaacctggc cattgatctg atgtacggag
     1081 gcatctactg ttttctgtgc caggactaca tctatgacaa agacatggaa ataatcgcca
     1141 aggaggagca gcgaaaagct tggaaaatgc aaggcgttgg agagaagttt tcaacttggg
     1201 aaccaaccaa acgggagctt gaactgctga agcacaaccc gaaaaggaga aagatcacct
     1261 cgaactgcac cataggtctg cgtgggctga tcaaccttgg gaacacatgc ttcatgaact
     1321 gcatcgtgca ggccctgacc cacacgccac ttctgcggga cttcttcctg tctgacaggc
     1381 accgctgtga gatgcagagc cccagctcct gtctggtctg tgagatgtcc tcactgtttc
     1441 aggagtttta ctctggacac cggtcccctc acatcccgta taagttgctg cacctggtgt
     1501 ggacccacgc gaggcaccta gcaggctacg agcagcagga cgcccacgag ttcctcatcg
     1561 cggccctgga cgtgctccac cgacactgca aaggtgatga caatgggaag aaggccaaca
     1621 accccaacca ctgcaactgc atcatagacc agatcttcac aggcgggttg cagtcagacg
     1681 tcacctgcca agtctgccat ggagtctcca ccaccatcga ccccttctgg gacatcagct
     1741 tggatctccc cggctcttcc accccattct ggcccctgag cccagggagc gagggcaacg
     1801 tggtaaacgg ggaaagccac gtgtcgggaa ccaccacgct cacggactgc ctgcgacgat
     1861 tcaccagacc agagcacttg ggcagcagcg ccaagatcaa gtgcagcggt tgccatagct
     1921 accaggagtc cacaaagcag ctcactatga agaaactgcc catcgtagcc tgttttcatc
     1981 tcaaacgatt tgaacactca gccaagctgc ggcggaagat caccacgtat gtgtccttcc
     2041 ccctggagct ggacatgacc cctttcatgg cctccagcaa agagagcagg atgaatggac
     2101 agtaccagca gcccacggac agtctcaaca atgacaacaa gtattccctg tttgctgttg
     2161 ttaaccatca agggaccttg gagagtggcc actacaccag ctttatccgg cagcacaaag
     2221 accagtggtt caagtgtgac gatgccatca tcaccaaggc cagcatcaag gacgtcctgg
     2281 acagcgaagg gtacttgctg ttctatcaca aacagttcct ggaatacgag ggatccgact
     2341 acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca
     2401 agtagtgagg gcccgatatc tcgacaatca acctctggat tacaaaattt gtgaaagatt
     2461 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     2521 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     2581 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     2641 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     2701 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     2761 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     2821 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     2881 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     2941 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     3001 ggccgcctcc ccgcctggaa acgggggagg ctaactgaaa cacggaagga gacaataccg
     3061 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc
     3121 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     3181 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     3241 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcagatct
     3301 gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3361 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3421 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3481 tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3541 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3601 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3661 ccctatctcg gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt
     3721 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3781 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3841 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3901 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3961 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4021 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     4081 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     4141 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     4201 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4261 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4321 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     4381 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     4441 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     4501 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     4561 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     4621 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     4681 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     4741 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     4801 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     4861 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     4921 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     4981 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5041 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5101 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     5161 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5221 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5281 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5341 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5401 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5461 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5521 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5581 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5641 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5701 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5761 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5821 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5881 ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt
     5941 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6001 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6061 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6121 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     6181 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6241 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6301 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6361 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6421 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6481 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6541 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6601 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6661 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6721 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6781 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6841 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6901 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6961 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7021 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7081 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7141 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7201 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7261 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7321 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7381 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7441 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtcg
     7501 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     7561 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     7621 accgacaatt gcatgaagaa tctgcttagg
//
