LOCUS       V046831                 7784 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046831
VERSION     V046831
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             908..3481
                     /label="USP5(NM_001098536)"
                     /note="USP5(NM_001098536)"
                     /gene="USP5"
     CDS             3482..3499
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    3546..3770
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3816..4244
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4258..4587
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4654..5253
                     /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    5427..5548
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5597..5613)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5621..5637
                     /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(5645..5675)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5690..5711
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5999..6584)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6755..7615)
                     /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(7616..7720)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg gcggagctga gtgaggaggc gctgctgtca gtattaccga cgatccgggt
      961 ccctaaggct ggagaccggg tccacaaaga cgagtgcgcc ttctccttcg acacgccgga
     1021 gtctgagggg ggcctctaca tctgtatgaa cacgtttctg ggctttggga aacagtatgt
     1081 ggagagacat ttcaataaga ccggccagcg agtctacttg cacctccggc ggacccggcg
     1141 cccgaaagag gaggaccctg ctacaggcac tggagaccca ccccggaaga agcccacgcg
     1201 gctggctatt ggtgttgaag gcggatttga ccttagcgag gagaagtttg aattagacga
     1261 ggatgtgaag attgtcattt tgccagatta cctggagatt gcccgggatg gactgggggg
     1321 actgcctgac attgtcagag atcgggtgac cagtgcagtg gaggccctac tgtcggccga
     1381 ctcagcctcc cgcaagcagg aggtgcaggc atgggatggg gaagtacggc aggtgtctaa
     1441 gcatgccttc agcctcaagc agttggacaa ccctgctcga atccctccct gtggctggaa
     1501 gtgctccaag tgtgacatga gagagaacct gtggctcaac ctgactgatg gctccatcct
     1561 ctgtgggcga cgctacttcg atggcagtgg gggcaacaac cacgctgtgg agcactaccg
     1621 agagacaggc tacccgttag ctgtcaagct gggcaccatc acccctgatg gagctgacgt
     1681 gtactcatat gatgaggatg acatggtcct ggaccccagc ctggctgagc acctgtccca
     1741 cttcggcatc gacatgctga agatgcagaa gacagacaag acgatgactg agttggagat
     1801 agacatgaac cagcggattg gtgaatggga gctgatccag gagtcaggtg tgccactcaa
     1861 gcccctgttt gggcctggct acacaggcat ccggaacctg ggtaacagct gctacctcaa
     1921 ctctgtggtc caggtgctct tcagcatccc tgacttccag aggaagtatg tggataagct
     1981 ggagaagatc ttccagaatg ccccgacgga ccctacccag gatttcagca cccaggtggc
     2041 caagctgggc catggccttc tctccgggga gtattccaag ccagtaccgg agtcgggcga
     2101 tggggagcgg gtgccagaac agaaggaagt tcaagatggc attgcccctc ggatgttcaa
     2161 ggccctcatc ggcaagggcc accctgaatt ctccaccaac cggcagcagg atgcccagga
     2221 gttcttcctt caccttatca acatggtgga gaggaattgc cggagctctg aaaatcctaa
     2281 tgaagtgttc cgcttcttgg tggaggaaaa gatcaagtgc ctggccacag agaaggtgaa
     2341 gtacacccag cgagttgact acatcatgca gctgcctgtg cccatggatg cagcccttaa
     2401 caaagaggag cttctggagt acgaggagaa gaagcggcaa gccgaagagg agaagatggc
     2461 actgccagaa ctggttcggg cccaggtgcc cttcagctct tgcctggagg cctacggggc
     2521 ccctgagcag gtcgatgact tctggagcac ggccctgcag gccaagtcag tagctgtcaa
     2581 gaccacacga tttgcctcat tccctgacta cctggtcatc cagatcaaga agttcacctt
     2641 cggcttagac tgggtgccca agaaactgga tgtgtccatc gagatgccag aggagctcga
     2701 catctcccag ttgaggggca cagggctgca gcccggagag gaggagctgc cagacattgc
     2761 cccacccctg gtcactccgg atgagcccaa aggtagcctt ggtttctatg gcaacgaaga
     2821 cgaagactcc ttctgctccc ctcacttctc ctctccgaca tcgcccatgc tggatgaatc
     2881 agtcatcatc cagctggtgg agatgggatt ccctatggac gcctgccgca aagctgtcta
     2941 ctacacgggc aacagcgggg ctgaggccgc catgaactgg gtcatgtcac acatggatga
     3001 tccagatttt gcaaaccccc tcatcctgcc tggctctagt gggccgggct ccacaagcgc
     3061 agcagccgac ccccctcctg aggactgtgt gaccaccatt gtctccatgg gcttctcccg
     3121 ggaccaggcc ttgaaagcgc tgcgggccac gaacaatagt ttagaacggg ctgtggactg
     3181 gatcttcagt cacattgacg acctggatgc tgaagctgcc atggacatct cagagggccg
     3241 ctcagctgcc gactccatct ctgagtctgt gccagtggga cctaaagtcc gggatggtcc
     3301 tggaaagtat cagctctttg ccttcattag tcacatgggc acctctacca tgtgtggtca
     3361 ctacgtctgc cacatcaaga aagaaggcag atgggtgatc tacaatgacc agaaagtgtg
     3421 tgcctccgag aagccgccca aggacctggg ctacatctac ttctaccaga gagtggccag
     3481 ccatcaccat caccatcact gactcgagtc tagagggccc gtttaaaccc gctgatcagc
     3541 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     3601 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     3661 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     3721 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     3781 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     3841 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3901 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3961 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4021 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4081 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4141 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     4201 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     4261 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4321 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4381 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4441 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4501 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     4561 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     4621 tctgatcaag agacaggatg aggatcgttt cgcatgaccg agtacaagcc cacggtgcgc
     4681 ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac
     4741 taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt caccgagctg
     4801 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     4861 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     4921 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     4981 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     5041 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     5101 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     5161 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     5221 acctggtgca tgacccgcaa gcccggtgcc tgagcgggac tctggggttc gaaatgaccg
     5281 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     5341 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5401 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     5461 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5521 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     5581 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     5641 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5701 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5761 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5821 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5881 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5941 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     6001 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6061 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6121 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6181 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6241 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6301 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6361 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6421 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6481 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt
     6541 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6601 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6661 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6721 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6781 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6841 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6901 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6961 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     7021 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     7081 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     7141 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     7201 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7261 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     7321 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     7381 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7441 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7501 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7561 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7621 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7681 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7741 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggt
//
