LOCUS       V019644                 7072 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019644
VERSION     V019644
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             928..1017
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     CDS             1051..2625
                     /label="USP22(NM_015276)"
                     /note="USP22(NM_015276)"
                     /gene="USP22"
     polyA_signal    2672..2896
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2942..3370
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3384..3713
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3780..4574
                     /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    4748..4869
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4918..4934)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4942..4958
                     /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(4966..4996)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5011..5032
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5320..5905)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6076..6936)
                     /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(6937..7041)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagccaccat ggcggcggaa caaaaactaa tatcagagga agatttggaa
      961 cagaaactaa tcagtgaaga agatttagag caaaagttaa tttccgagga ggacttatca
     1021 gcggaaaagc ttggtaccga gctcggatcc atggtgtccc ggccagagcc cgagggcgag
     1081 gccatggacg ccgagctggc ggtagcgccg ccgggctgct cgcacctggg cagcttcaag
     1141 gtggacaact ggaagcagaa cctgcgggcc atctaccagt gcttcgtgtg gagcggcacg
     1201 gctgaggccc gcaagcgcaa ggccaagtcc tgtatctgcc atgtctgtgg cgtccacctc
     1261 aacaggctgc attcctgcct ctactgtgtc ttcttcggct gtttcacaaa gaagcatatt
     1321 cacgagcatg cgaaggcgaa gcggcacaac ctggccattg atctgatgta cggaggcatc
     1381 tactgttttc tgtgccagga ctacatctat gacaaagaca tggaaataat cgccaaggag
     1441 gagcagcgaa aagcttggaa aatgcaaggc gttggagaga agttttcaac ttgggaacca
     1501 accaaacggg agcttgaact gctgaagcac aacccgaaaa ggagaaagat cacctcgaac
     1561 tgcaccatag gtctgcgtgg gctgatcaac cttgggaaca catgcttcat gaactgcatc
     1621 gtgcaggccc tgacccacac gccacttctg cgggacttct tcctgtctga caggcaccgc
     1681 tgtgagatgc agagccccag ctcctgtctg gtctgtgaga tgtcctcact gtttcaggag
     1741 ttttactctg gacaccggtc ccctcacatc ccgtataagt tgctgcacct ggtgtggacc
     1801 cacgcgaggc acctagcagg ctacgagcag caggacgccc acgagttcct catcgcggcc
     1861 ctggacgtgc tccaccgaca ctgcaaaggt gatgacaatg ggaagaaggc caacaacccc
     1921 aaccactgca actgcatcat agaccagatc ttcacaggcg ggttgcagtc agacgtcacc
     1981 tgccaagtct gccatggagt ctccaccacc atcgacccct tctgggacat cagcttggat
     2041 ctccccggct cttccacccc attctggccc ctgagcccag ggagcgaggg caacgtggta
     2101 aacggggaaa gccacgtgtc gggaaccacc acgctcacgg actgcctgcg acgattcacc
     2161 agaccagagc acttgggcag cagcgccaag atcaagtgca gcggttgcca tagctaccag
     2221 gagtccacaa agcagctcac tatgaagaaa ctgcccatcg tagcctgttt tcatctcaaa
     2281 cgatttgaac actcagccaa gctgcggcgg aagatcacca cgtatgtgtc cttccccctg
     2341 gagctggaca tgaccccttt catggcctcc agcaaagaga gcaggatgaa tggacagtac
     2401 cagcagccca cggacagtct caacaatgac aacaagtatt ccctgtttgc tgttgttaac
     2461 catcaaggga ccttggagag tggccactac accagcttta tccggcagca caaagaccag
     2521 tggttcaagt gtgacgatgc catcatcacc aaggccagca tcaaggacgt cctggacagc
     2581 gaagggtact tgctgttcta tcacaaacag ttcctggaat acgagtagct cgagtctaga
     2641 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     2701 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2761 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     2821 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     2881 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     2941 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3001 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3061 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3121 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3181 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3241 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3301 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3361 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3421 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3481 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3541 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3601 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     3661 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     3721 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     3781 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     3841 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     3901 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     3961 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4021 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4081 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4141 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4201 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4261 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     4321 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4381 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4441 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4501 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4561 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     4621 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4681 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4741 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4801 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4861 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     4921 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4981 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5041 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5101 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5161 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5221 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5281 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5341 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5401 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5461 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5521 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5581 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5641 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5701 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5761 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5821 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat
     5881 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5941 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6001 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6061 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6121 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6181 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6241 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6301 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6361 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6421 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6481 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6541 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6601 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6661 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6721 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6781 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6841 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6901 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6961 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7021 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
