LOCUS       V033183                 8427 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033183
VERSION     V033183
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..3107
                     /label="Usp1(NM_146144)"
                     /note="Usp1(NM_146144)"
                     /gene="Usp1"
     CDS             3114..3179
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3203..3791
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3857..3905
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4107..4535
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4549..4878
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4945..5739
                     /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    5915..6036
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6085..6101)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6109..6125
                     /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(6133..6163)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6178..6199
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6487..7075)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7246..8106)
                     /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(8107..8211)
                     /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 ccaccatgcc tggcgtcata cctagtgaaa
      781 gtaatgggct ttcaagaggc agtccatcca agaaaaacag actgtctctg aagttttttc
      841 agaaaaagga aactaaaaga gccttagact ttactgattc tcaagaaaat gaagaaaaaa
      901 cttctgaata tagaggatct gagattgacc aagttgttcc tgctgcacag tcctccccag
      961 taagctgcga gaagagagag aacttgttac catttgtggg attgaataac cttggcaaca
     1021 cttgttatct gaatagtatt cttcaggtgt tatacttttg tcctggcttt aagactggag
     1081 tgaagcactt atttaatatt atttcaagga agaaagaagc tctgaaggat gactctaatc
     1141 agaaggataa gggaagctgc aaagaagagt ctttggcaag ttatgagctt atatgtagtt
     1201 tacagtcctt aataatttca gttgaacagc ttcaggctag ttttctctta aatccagaga
     1261 aatatacaga tgaacttgct acacagccaa ggcgactgct gaacacactc agggaactca
     1321 accctatgta tgaaggattt ctacagcatg atgcacagga agtgctacag tgtattctgg
     1381 gaaacattca agaaacatgc caactcctaa aaaaagaaga aataaaaaac ctggctgaat
     1441 tatccggcaa ggttgaagaa caatctcttc agaaagagga aacaggtgga attaccagca
     1501 cagagattga cagtatgagg aatacggagg acgtcaaaga gcagctcccc aaagggaact
     1561 ggaaaagaaa aagtgacagt gagtcgagta acgtgaagaa aaaggttaaa ctgtccaggg
     1621 aatcccagcc attggaagaa aaccagagac aaaccagatc caagagaaaa gctatcggag
     1681 acacactaga ggctgctcct aagatcatcc ccaagtgtgt ttctgaaagt gagagtgcaa
     1741 agccctcaca gaagaaatca aaagttaaaa taaattggtt aaagccatcc actaagcaac
     1801 ccagcattct ttccaagttc tgtagtcttg gaaaaataac aactaaccag cgctccaaag
     1861 gacaacctaa ggaaaaggag ggtgatgttg aagaggactt ggagaagtac ggaagtgacc
     1921 acacagctaa tggtggaccc gaatctccag gaagtagcgt cacacctgtg gacagcagtg
     1981 aagccaagtc cggaaacaaa ggtgctgaac agattggctt tgagctagta gagaaactgt
     2041 ttcaaggtca gctcgtgtta aggactcggt gtttggaatg tgaaagctta acagaaagaa
     2101 gagaagattt ccaggatatt agtgttccag tgcaagaaga tgagctttcc aaggtcgagg
     2161 agagctctga aatttctcca gagccaaaaa cagaaatgaa gaccttgaga tgggcaattt
     2221 cacagtttgc ttcagtggag agaattgtag gagaggataa atacttctgt gaaaattgcc
     2281 atcattatac ggaagcagag cgaagtctcc tgtttgataa aatgcctgaa gttatcacta
     2341 ttcatctgaa gtgctttgct gctagtggct tggagtttga ctgttacggt ggtggacttt
     2401 ccaagatcaa cactccttta ttgacacctc ttaaactgtc actagaagaa tggagcacaa
     2461 agccgacgaa tgacagctat ggattatttg ccgttgtgat gcacagtggc attactatta
     2521 gtagtgggca ctataccgca tctgttaaag tcactgacct taacagtcta gagctcgatg
     2581 aggggaattt tgtggttgac cagatgtgtg aactaggtaa gccagagcca ctgaccgagg
     2641 agcaagcaag ggggacggct gggaactacg acgatgaagt atcaattaga gttggtggaa
     2701 atgcccagcc aagtaaagtt ttgaacaaaa aaaatgtaga aggtattgga cttctcggtg
     2761 gacaaaagag caaagcagat tatgagctgt acaacaaagc atctaatcct gataaggtgg
     2821 ttgggacacc attcaccgac aatagaaatt ctgaaactaa tgatactact aatgggaccc
     2881 acgaatctga caggaacaag gaatccagtg accaaacagg cgttaacatg aacggacttg
     2941 agaacaaaat ttcctatgta gtgcaaagct taaaggagta cgaggggaag tggttgctct
     3001 ttgacgattc tgaggtaaaa gtgacagaag agaaagactt tctgaattct ctctcccctt
     3061 ctacatcccc tacgtctact ccctacttgc tattctataa aaaattagga tccgactaca
     3121 aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt
     3181 agtgagggcc cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     3241 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3301 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3361 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3421 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3481 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3541 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     3601 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     3661 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3721 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3781 cgcctccccg cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa
     3841 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt
     3901 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     3961 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     4021 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg
     4081 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     4141 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4201 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4261 gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4321 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4381 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4441 tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct attggttaaa
     4501 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     4561 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4621 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4681 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4741 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4801 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4861 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     4921 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4981 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     5041 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     5101 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     5161 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     5221 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     5281 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     5341 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5401 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5461 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5521 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5581 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5641 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5701 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga
     5761 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5821 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5881 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     5941 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     6001 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     6061 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     6121 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     6181 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6241 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6301 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6361 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6421 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6481 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6541 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6601 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6661 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6721 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6781 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6841 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6901 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     6961 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7021 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7081 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7141 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7201 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7261 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7321 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7381 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7441 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7501 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7561 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7621 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7681 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7741 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7801 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7861 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7921 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7981 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8041 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8101 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8161 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8221 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact
     8281 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     8341 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     8401 gacaattgca tgaagaatct gcttagg
//
