LOCUS       V045259                 8062 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045259
VERSION     V045259
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"
     CDS             925..3813
                     /label="USP4(NM_003363)"
                     /note="USP4(NM_003363)"
                     /gene="USP4"
     CDS             3826..3891
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3926..4150
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4196..4624
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4638..4967
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4979..5578
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    5738..5859
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5908..5924)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5932..5948
                     /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(5956..5986)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6001..6022
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6310..6895)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7066..7926)
                     /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(7927..8031)
                     /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 aagctttcta gagaattcgc caccatggcg gaaggtggag gctgccgtga gcgaccggat
      961 gcggagactc agaagtccga gcttggaccc ttaatgagga ccacactcca acgcggggcg
     1021 cagtggtatc ttattgacag ccggtggttc aagcagtgga agaagtatgt gggctttgac
     1081 agctgggaca tgtacaatgt gggtgaacat aacctatttc ctggcccaat agacaactct
     1141 gggctatttt cagatcctga gagtcagacc ttgaaagaac acttaattga tgaattggac
     1201 tatgtattgg tccctaccga ggcgtggaat aaactactaa actggtacgg ctgtgtagaa
     1261 ggccagcaac ccatcgtcag aaaagttgtg gagcatggcc tgtttgtcaa gcactgcaaa
     1321 gtcgaggtgt atttgctgga actgaagctc tgtgagaaca gtgaccccac caatgtgctg
     1381 agttgccatt tcagcaaggc agacaccatt gcaaccatcg agaaagagat gcggaagcta
     1441 ttcaacatcc ctgcggagcg tgaaacacgg ctctggaaca aatacatgag caacacctac
     1501 gagcagttga gcaagctaga caacactgtc caggatgctg ggctatacca gggtcaggtg
     1561 ctagtaattg agcctcaaaa tgaagatggc acatggccca ggcagacctt gcagtcaaaa
     1621 tcaagcactg cgcctagcag aaattttact acctctccaa aatcatcagc aagtccctat
     1681 tcctcagtgt ctgcctctct cattgcaaat ggtgatagca ctagcacctg tgggatgcac
     1741 agttccggtg tcagcagggg tggatctggc ttttctgctt cgtataattg tcaggagcca
     1801 ccatcctctc atatacaacc tgggctctgt ggacttggaa acctgggaaa cacctgcttc
     1861 atgaactccg ctttgcagtg tttgagcaac actgcaccac tgactgacta ctttctcaaa
     1921 gatgagtatg aagccgaaat caacagagac aaccctctgg ggatgaaagg ggaaattgca
     1981 gaagcctatg ctgaactcat taagcagatg tggtctggaa gggacgccca tgtggcacct
     2041 cgcatgttca aaactcaagt aggacgtttt gctcctcaat tttctggcta ccagcaacaa
     2101 gattctcagg agctgctggc ctttcttcta gatggattgc atgaagatct gaaccgggta
     2161 aagaaaaagc cctacttgga gctgaaggat gccaatgggc ggccagatgc ggtggtggca
     2221 aaggaagcct gggagaatca caggttgagg aatgattctg tgattgtgga tactttccat
     2281 ggcctcttca aatctacttt ggtttgccca gaatgtgcta aggtttctgt gacctttgac
     2341 ccattttgct atctaacgct gccactgccc ttgaagaaag atcgagttat ggaggttttc
     2401 ctggttcctg ctgaccctca ctgcagacct actcagtacc gtgtgactgt gccgctgatg
     2461 ggggctgtgt ccgacctgtg cgaggctctc tccaggctgt ctggcattgc tgcagaaaat
     2521 atggtggtcg cagatgtgta taatcaccga ttccacaaaa ttttccaaat ggatgaaggt
     2581 ttaaaccaca tcatgcctcg ggatgacatt ttcgtgtacg aggtctgcag cacttccgtg
     2641 gatggctcgg aatgtgtcac gcttccagtc tacttcaggg agaggaagtc caggccatca
     2701 agcacttcct ccgcatcagc gctatatggg cagccactat tgctttctgt ccccaagcac
     2761 aagttaaccc ttgagtcttt gtaccaggct gtttgtgatc gtatcagccg ctatgtgaaa
     2821 cagcctttac ctgatgagtt tggcagctca cccttggagc caggggcctg caatggctcc
     2881 aggaacagct gtgaaggaga agatgaggaa gaaatggagc atcaggaaga aggcaaagag
     2941 cagctttcag aaacagaagg cagtggggaa gatgagccag gaaatgaccc cagtgagacc
     3001 acccaaaaga agatcaaagg ccagccctgc ccaaaaaggc tttttacctt cagtcttgtg
     3061 aactcctatg gaacagctga cataaattca cttgcagctg atggaaaact acttaaactc
     3121 aactctcgat ctacactggc catggattgg gacagtgaaa ctcggagact ttactatgat
     3181 gagcaagaat ctgaggccta cgagaagcat gtgagcatgt tgcagcctca gaagaagaag
     3241 aagaccacag tggccctgag agactgcatc gagctcttca ccaccatgga gacccttggg
     3301 gagcatgacc cctggtactg tcccaactgt aagaagcatc aacaggccac aaaaaagttt
     3361 gacctatggt ccttgcccaa gatcctggtg gtccacctca aacgtttctc ctacaacaga
     3421 tactggaggg ataagctcga cacagtcgta gaattcccaa tcagagggct gaacatgtcc
     3481 gagtttgtct gtaacctgtc agcaaggcct tatgtgtacg acctcattgc cgtgtccaat
     3541 cattatggag ccatgggggt tggccactac actgcatatg cgaagaacaa actgaatggt
     3601 aaatggtatt actttgatga tagcaacgtg tccctggcct ctgaggatca gatagtgact
     3661 aaagcagctt atgtgctatt ttaccaacgt cgagatgatg aattttataa gacaccttca
     3721 cttagcagtt ctggttcctc tgatggaggg acacgaccaa gcagctctca gcagggcttt
     3781 ggggatgatg aggcttgcag catggacacc aacctcgaga ccggtgacta caaagaccat
     3841 gacggtgatt ataaagatca tgacatcgat tacaaggatg acgatgacaa gtgaccggta
     3901 agtttaaacc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     3961 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     4021 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     4081 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     4141 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     4201 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4261 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4321 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4381 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     4441 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4501 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     4561 attttggcca tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4621 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     4681 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4741 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4801 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4861 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     4921 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggat
     4981 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
     5041 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
     5101 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     5161 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     5221 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     5281 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     5341 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     5401 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     5461 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     5521 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgatt
     5581 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5641 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5701 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     5761 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5821 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5881 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5941 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     6001 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6061 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6121 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6181 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6241 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6301 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6361 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6421 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6481 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6541 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6601 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6661 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6721 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6781 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6841 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6901 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6961 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7021 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7081 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7141 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7201 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7261 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7321 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7381 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7441 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7501 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7561 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7621 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7681 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7741 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7801 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7861 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7921 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7981 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8041 ccgaaaagtg ccacctgacg tc
//
