LOCUS       V019962                 8852 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019962
VERSION     V019962
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             683..3508
                     /label="USP33(NM_015017)"
                     /note="USP33(NM_015017)"
                     /gene="USP33"
     CDS             3515..3604
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    3628..4216
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4282..4330
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4532..4960
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4974..5303
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5370..6164
                     /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    6340..6461
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6510..6526)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6534..6550
                     /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(6558..6588)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6603..6624
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6912..7500)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7671..8531)
                     /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(8532..8636)
                     /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 ggatccgcca ccatgacagg atcaaattca cacataacga tattaacctt
      721 aaaggtgtta cctcattttg aaagtcttgg gaaacaggaa aaaattccta acaaaatgtc
      781 agcttttcga aatcattgtc cacatttgga ttcagttggt gaaataacaa aagaagattt
      841 gatacaaaaa tcccttggta cttgtcagga ttgtaaagtc caaggaccaa atctttgggc
      901 atgtctggag aatagatgtt catatgttgg ctgtggtgaa tcacaagtag atcacagcac
      961 catacattct caggagacaa agcattatct aactgtgaac cttaccactc ttcgagtatg
     1021 gtgttatgct tgcagcaaag aagtattttt ggataggaaa ttaggaactc agccttcatt
     1081 gcctcatgta agacaacctc accaaataca agaaaacagt gtccaggatt ttaaaatacc
     1141 cagtaataca acattaaaaa ctcctctggt tgccgtattt gatgatctgg atatagaagc
     1201 ggatgaagaa gatgaactta gggccagagg tcttacaggt ttgaaaaata ttggaaatac
     1261 ttgttacatg aatgcagctt tgcaggctct ttctaattgc ccacctttga cacagttttt
     1321 tcttgattgt ggaggactag ctcgaacaga taagaaacct gccatttgta aaagttatct
     1381 caaactaatg acagagctgt ggcataaaag caggccagga tctgttgtgc ctactactct
     1441 gtttcaagga attaaaactg taaatccaac atttcggggg tattctcagc aggatgctca
     1501 agaattcctt cgatgtttaa tggatttgct tcatgaagaa ttgaaagagc aagtcatgga
     1561 agtagaagaa gatccgcaaa ccataaccac tgaggagaca atggaagaag acaagagcca
     1621 gtcggatgta gattttcagt cttgtgaatc ttgtagcaac agtgatagag cagaaaatga
     1681 aaatggctct agatgctttt ctgaagataa taatgaaaca acaatgttaa ttcaggatga
     1741 tgaaaacaat tcagaaatgt caaaggattg gcaaaaagag aagatgtgca ataagattaa
     1801 taaagtaaat tctgaaggcg aatttgataa agatagagac tctatatctg aaacagtcga
     1861 cttaaacaac caggaaactg tcaaagtgca aatacacagc agagcttcag aatatatcac
     1921 tgatgtccat tcgaatgacc tgtctacacc acagatcctt ccatcaaatg aaggtgttaa
     1981 tccacgttta tcggcaagcc ctcctaaatc aggcaatttg tggccaggat tggcaccacc
     2041 acacaaaaaa gctcagtctg catctccaaa gagaaaaaaa cagcacaaga aatacagaag
     2101 tgttatttca gacatatttg atggaacaat cattagttca gtgcagtgtc tgacttgtga
     2161 cagggtgtct gtaaccctcg agacctttca agatctgtcc ttgccaattc ctggcaagga
     2221 agaccttgct aagctgcatt catcaagtca tccaacttct atagtcaaag caggatcatg
     2281 tggcgaagca tatgctccac aagggtggat agcttttttc atggaatatg tgaagaggtt
     2341 tgttgtctca tgtgtcccta gctggttttg gggtccagta gtaaccttgc aagattgtct
     2401 tgctgccttc tttgccagag atgaactaaa aggtgacaat atgtacagtt gtgaaaaatg
     2461 caaaaagttg agaaatggag tgaagttttg taaagtacaa aactttcctg agattttgtg
     2521 catccacctt aaaagattca gacatgaact aatgttttcc accaaaatca gtacccatgt
     2581 ttcatttccg ctagaaggct tggatcttca gccatttctt gctaaggata gtccagctca
     2641 aattgtgaca tatgatcttc tgtcagtcat ttgccatcat ggaactgcaa gtagtggaca
     2701 ctatatagcc tactgccgaa acaatctaaa taatctctgg tatgaatttg atgatcagag
     2761 tgtcactgaa gtttcagaat ctactgtaca aaatgcagaa gcttacgttc ttttctatag
     2821 gaagagcagc gaagaggcac aaaaagagag gagaaggata tcaaatttat tgaacataat
     2881 ggaaccaagc ctccttcagt tttatatttc tcgacagtgg cttaataaat ttaagacctt
     2941 tgccgaacct ggccctattt caaataatga ctttctttgt attcatggag gtgttcctcc
     3001 aagaaaagct ggttatattg aagacctggt tttgatgctg cctcagaaca tttgggataa
     3061 cctatatagc aggtatggtg gaggaccagc tgtcaaccat ctgtacattt gtcatacttg
     3121 ccaaattgag gcggagaaaa ttgaaaaaag aagaaaaact gaattggaaa tttttattcg
     3181 gcttaacaga gcgttccaaa aagaggactc tccagctact ttttattgca tcagtatgca
     3241 gtggtttaga gaatgggaaa gttttgtgaa gggtaaagat ggagatcctc caggtcctat
     3301 tgacaatact aagattgcag tcactaaatg tggtaatgtg atgcttaggc aaggagcaga
     3361 ttctggccag atttctgaag aaacatggaa ttttctgcag tctatttatg gtggagggcc
     3421 tgaagttatc ctgcgacctc cggttgttca tgttgatcca gatatacttc aagcagaaga
     3481 aaaaattgaa gtagaaactc ggtctttgct cgaggaacaa aaactaatat cagaggaaga
     3541 tttggaacag aaactaatca gtgaagaaga tttagagcaa aagttaattt ccgaggagga
     3601 cttatagtga gggcccgata tctcgacaat caacctctgg attacaaaat ttgtgaaaga
     3661 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     3721 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     3781 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     3841 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     3901 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     3961 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     4021 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     4081 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     4141 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     4201 tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag gagacaatac
     4261 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac gggtgttggg
     4321 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     4381 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     4441 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcagat
     4501 ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     4561 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4621 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4681 aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4741 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4801 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4861 aaccctatct cggtctattc ttttgattta taagggattt tggggatttc ggcctattgg
     4921 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     4981 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5041 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5101 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     5161 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     5221 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     5281 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     5341 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     5401 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     5461 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     5521 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     5581 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     5641 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     5701 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     5761 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5821 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     5881 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5941 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     6001 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     6061 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     6121 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     6181 cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     6241 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     6301 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     6361 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     6421 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     6481 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     6541 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     6601 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     6661 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     6721 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     6781 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6841 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6901 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6961 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     7021 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     7081 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     7141 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     7201 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     7261 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     7321 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     7381 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7441 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7501 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7561 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     7621 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     7681 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     7741 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7801 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7861 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7921 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7981 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     8041 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     8101 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     8161 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     8221 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     8281 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     8341 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     8401 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     8461 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     8521 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     8581 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     8641 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     8701 cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     8761 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     8821 tgaccgacaa ttgcatgaag aatctgctta gg
//
