LOCUS       V019633                 9398 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019633
VERSION     V019633
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..4054
                     /label="USP36(NM_001385169)"
                     /note="USP36(NM_001385169)"
                     /gene="USP36"
     CDS             4061..4150
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    4174..4762
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4828..4876
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5078..5506
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5520..5849
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5916..6710
                     /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    6886..7007
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7056..7072)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7080..7096
                     /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(7104..7134)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7149..7170
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7458..8046)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8217..9077)
                     /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(9078..9182)
                     /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 ccatgccaat agtggataag ttgaaggagg ccctgaaacc
      721 cggccgcaag gactcggctg atgatggaga actggggaag cttcttgcct cctctgccaa
      781 gaaggtcctt ttacagaaaa tcgagttcga gccagccagc aagagcttct cctaccagct
      841 ggaggcctta aagagcaaat atgtgttgct caaccccaaa acagagggag ctagtcgcca
      901 caagagtgga gatgacccac cggccaggag acagggcagt gagcacacgt atgagagctg
      961 tggtgacgga gtcccagccc cgcagaaagt gcttttcccc acggagcgac tgtctctgag
     1021 gtgggagcgg gtcttccgcg tgggcgcagg actccacaac cttggcaaca cctgctttct
     1081 caatgccacc atccagtgct tgacctacac accacctcta gccaactacc tgctctccaa
     1141 ggagcatgct cgcagctgcc accagggaag cttctgcatg ctgtgtgtca tgcagaacca
     1201 cattgtccag gccttcgcca acagcggcaa cgccatcaag cccgtctcct tcatccgaga
     1261 cctgaaaaag atcgcccgac acttccgctt tgggaaccag gaggacgcgc atgagttcct
     1321 gcggtacacc atcgacgcca tgcagaaagc ctgcctgaat ggctgtgcca agttggatcg
     1381 tcaaacgcag gctactacct tggtccatca aatttttgga gggtatctca gatcacgcgt
     1441 gaagtgctcc gtgtgcaaga gcgtctcgga cacctacgac ccctacttgg acgtcgcgct
     1501 ggagatccgg caagctgcga atattgtgcg tgctctggaa ctttttgtga aagcagatgt
     1561 cctgagtgga gagaatgcct acatgtgtgc taaatgcaag aagaaggttc cagccagcaa
     1621 gcgcttcacc atccacagaa catccaacgt cttaaccctt tccctcaagc gctttgccaa
     1681 cttcagcggg gggaagatca ccaaggatgt aggctatccg gaattcctca acatacgtcc
     1741 gtatatgtcc cagaataatg gtgatcctgt catgtatgga ctctatgctg tcctggtgca
     1801 ctcgggctac agctgccatg ccgggcacta ttactgctac gtgaaggcaa gcaatggaca
     1861 gtggtaccag atgaatgatt ccttggtcca ttccagcaac gtcaaggtgg ttctgaacca
     1921 gcaggcctac gtgctgttct atctgcgaat tccaggctct aagaaaagtc ccgagggcct
     1981 catctccagg acaggctcct cctcccttcc cggccgcccg agtgtgattc cagatcactc
     2041 caagaagaac atcggcaatg ggattatttc ctccccactg actggaaagc gacaagactc
     2101 tgggacgatg aagaagccgc acaccactga agagattggt gtgcccatat ccaggaatgg
     2161 ctccaccctg ggcctgaagt cccagaacgg ctgcattcct ccaaagctgc cctcggggtc
     2221 cccttccccc aaactctccc agacacccac acacatgcca accatcctag acgaccctgg
     2281 aaagaaggtg aagaagccag ctcctccaca gcacttttcc cccagaactg ctcaggggct
     2341 gcctgggacc agcaactcga atagcagcag atctgggagc caaaggcagg gctcctggga
     2401 cagcagggat gttgtcctct ctacctcacc taagctcctg gctacagcca ctgccaacgg
     2461 gcatgggctg aaggggaacg acgagagcgc tggcctcgac aggaggggct ccagcagctc
     2521 cagcccagag cactcggcca gcagcgactc caccaaggcc ccccagaccc ccaggagtgg
     2581 agcggcccat ctctgcgatt ctcaggaaac gaactgttcc accgctggcc actccaaaac
     2641 gccgccaagt ggagcagatt ctaagacggt gaagctgaag tcccctgtcc tgagcaacac
     2701 caccactgag cctgcaagca ccatgtctcc tccaccagcc aaaaaactgg ccctttctgc
     2761 caagaaggcc agcaccctgt ggagggcgac cggcaatgac ctccgtccac ctcccccctc
     2821 accatcctcc gacctcaccc accccatgaa aacctctcac cccgtcgttg cctccacttg
     2881 gcccgtccat agagccaggg ctgtgtcacc tgctccccaa tcatccagcc gcctgcaacc
     2941 ccccttcagc ccccacccca cattgctgtc cagtaccccc aagcccccag ggacgtcaga
     3001 accacggagc tgctcctcca tctcgacggc gctgcctcag gtcaacgagg accttgtgtc
     3061 tcttccacac cagttgccag aggccagtga gcccccccag agcccctctg agaagaggaa
     3121 aaagaccttt gtgggagagc cgcagaggct gggctcagag acgcgcctcc cacagcacat
     3181 cagggaggcc actgcggctc cccacgggaa gaggaagagg aagaagaaga agcgcccgga
     3241 ggacacagct gccagcgccc tgcaggaggg gcagacacag agacagcctg ggagccccat
     3301 gtacaggagg gagggccagg cacagctgcc cgctgtcaga cggcaggaag atggcacaca
     3361 gccacaggtg aatggccagc aggtgggatg tgttacggac ggccaccacg cgagcagcag
     3421 gaagcggagg aggaaaggag cagaaggtct tggtgaagaa ggcggcctgc accaggaccc
     3481 acttcggcac agcagctgct ctcccatggg tgatggtgat ccagaggcca tggaagagtc
     3541 tccaaggaaa aagaaaaaga aaaaaagaaa gcaggagaca cagcgggcag tagaagagga
     3601 tgggcatctc aaatgcccaa ggagtgccaa gccccaagat gctgttgtcc ccgagtccag
     3661 cagctgcgca ccatccgcga atggctggtg tcctggggac cgcatggggc tgagccaggc
     3721 ccctcctgtg tcttggaatg gagagcggga gtctgatgtg gtccaggaac tgctcaaata
     3781 ctcatctgat aaagcttacg ggagaaaagt tctgacctgg gatggcaaga tgtcggcggt
     3841 cagtcaggat gctattgaag acagcagaca ggcccggact gagaccgtgg ttgatgactg
     3901 ggacgaagag tttgaccgag ggaaggaaaa gaaaattaaa aaatttaaga gagagaagag
     3961 gagaaacttc aacgccttcc agaaacttca gactcgacgg aacttctggt ctgtgactca
     4021 cccagcaaag gctgccagcc tcagctatcg ccgcctcgag gaacaaaaac taatatcaga
     4081 ggaagatttg gaacagaaac taatcagtga agaagattta gagcaaaagt taatttccga
     4141 ggaggactta tagtgagggc ccgatatctc gacaatcaac ctctggatta caaaatttgt
     4201 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     4261 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     4321 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     4381 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     4441 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     4501 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     4561 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
     4621 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     4681 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     4741 tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca cggaaggaga
     4801 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacgggt
     4861 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     4921 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     4981 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     5041 gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     5101 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     5161 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     5221 gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     5281 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     5341 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     5401 acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc
     5461 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     5521 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     5581 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     5641 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     5701 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     5761 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     5821 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     5881 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     5941 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     6001 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     6061 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     6121 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     6181 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     6241 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     6301 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     6361 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     6421 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     6481 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     6541 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     6601 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     6661 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     6721 ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     6781 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     6841 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     6901 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     6961 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     7021 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     7081 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     7141 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     7201 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     7261 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     7321 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     7381 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     7441 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     7501 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     7561 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     7621 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg
     7681 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     7741 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     7801 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7861 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     7921 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7981 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     8041 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     8101 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     8161 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     8221 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     8281 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     8341 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     8401 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     8461 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     8521 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     8581 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     8641 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     8701 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     8761 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     8821 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     8881 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     8941 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     9001 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     9061 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     9121 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     9181 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     9241 tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     9301 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     9361 aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
