LOCUS       V040375                 7828 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040375
VERSION     V040375
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             964..3318
                     /label="USP1(NM_003368)"
                     /note="USP1(NM_003368)"
                     /gene="USP1"
     CDS             3325..3390
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3428..3652
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3698..4126
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4140..4469
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4536..5330
                     /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    5504..5625
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5674..5690)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5698..5714
                     /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(5722..5752)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5767..5788
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6076..6661)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6832..7692)
                     /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(7693..7797)
                     /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 aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatgcctg gtgtcatacc tagtgaaagt aatggacttt caagaggtag cccttcaaag
     1021 aaaaacagac tttccttaaa gttttttcag aaaaaggaaa ctaagagagc tttggatttc
     1081 acagattctc aagaaaatga agaaaaagct tctgaatata gagcatctga aattgatcaa
     1141 gttgttcctg cagcacagtc ttcacctata aactgtgaga agagagaaaa cttgttacca
     1201 tttgtgggac tgaataatct cggcaatact tgctatctta atagtatact tcaggtatta
     1261 tatttttgtc ccggttttaa atctggagta aagcacttat ttaatattat ttcaaggaag
     1321 aaagaagctc taaaggatga agccaatcaa aaagacaagg gaaattgcaa agaagattct
     1381 ttggcaagtt atgaattgat atgcagttta cagtccttaa tcatttcggt tgaacagctc
     1441 caggctagtt ttctcttaaa tccagagaaa tatactgatg aacttgccac tcagccaagg
     1501 cgactgctta acacactgag ggaactcaac cctatgtatg aaggatatct acagcatgat
     1561 gcacaggaag tattacaatg tattttggga aacattcaag aaacatgcca actcctaaaa
     1621 aaagaagaag taaaaaatgt ggcagaatta cctactaagg tagaagaaat acctcatccg
     1681 aaagaggaaa tgaatggtat taacagcata gagatggaca gtatgaggca ttctgaagac
     1741 tttaaagaga aactcccaaa aggaaatggg aaaagaaaaa gtgacactga atttggtaac
     1801 atgaagaaaa aagttaaatt atccaaggaa caccagtcat tggaagagaa ccagagacaa
     1861 actagatcaa aaagaaaagc tacaagtgat acattagaga gtcctcctaa aataattccc
     1921 aagtatattt ctgaaaatga gagtccaaga ccctcacaaa agaaatcaag agttaaaata
     1981 aattggttaa agtctgcaac taagcaaccc agcattcttt ctaaattttg tagtctggga
     2041 aaaataacaa caaaccaagg agtcaaagga caatctaaag aaaatgaatg tgatcctgaa
     2101 gaggacttgg ggaagtgtga aagtgataac acaactaatg gttgtggact tgaatctcca
     2161 ggaaatactg ttacacctgt aaatgttaat gaagttaaac ccataaacaa aggtgaagaa
     2221 caaattggtt ttgagctagt ggagaaatta tttcaaggtc agctggtatt aaggacgcgt
     2281 tgcttggaat gtgaaagttt aacagaaaga agagaagatt ttcaagacat cagtgtgcca
     2341 gtacaagaag atgagctttc caaagtagag gagagttctg aaatttctcc agagccaaaa
     2401 acagaaatga agaccctgag atgggcaatt tcacaatttg cttcagtaga aaggattgta
     2461 ggagaagata aatatttctg tgaaaactgc catcattata ctgaagctga acgaagtctt
     2521 ttgtttgaca aaatgcctga agttataact attcatttga agtgctttgc tgctagtggt
     2581 ttggagtttg attgttatgg tggtggactt tccaagatca acactccttt attgacacct
     2641 cttaaattgt cactagaaga atggagcaca aagccaacta acgacagcta tggattattt
     2701 gcggttgtga tgcatagtgg cattacaatt agtagtgggc attacactgc ttctgttaaa
     2761 gtcactgacc ttaacagttt agaactagat aaaggaaatt ttgtggttga ccaaatgtgt
     2821 gaaataggta agccagaacc attgaatgag gaggaagcaa ggggtgtggt tgagaattat
     2881 aatgatgaag aagtgtcaat tagagttggt ggaaatacac agccaagtaa agttttgaac
     2941 aaaaaaaatg tagaagctat tggacttctt ggaggacaaa agagcaaagc agattatgag
     3001 ctatacaaca aagcctctaa tcctgataag gttgctagta cagcgtttgc tgaaaataga
     3061 aattctgaga ctagtgatac tactgggacc catgaatctg atagaaacaa ggaatccagt
     3121 gaccaaacag gcattaatat tagtggattt gagaacaaaa tttcatacgt agtgcaaagc
     3181 ttaaaggagt atgaggggaa gtggttgctt tttgatgatt ctgaagtcaa agttactgaa
     3241 gagaaggact ttctgaattc tctttcccct tctacatctc ctacttctac tccttacttg
     3301 ctattttata agaaattact cgaggactac aaagaccatg acggtgatta taaagatcat
     3361 gacatcgact acaaggatga cgatgacaag tagtgagggc ccgtttaaac ccgctgatca
     3421 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     3481 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     3541 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     3601 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     3661 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     3721 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3781 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3841 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3901 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3961 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4021 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     4081 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     4141 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4201 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4261 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4321 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4381 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     4441 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     4501 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     4561 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4621 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4681 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     4741 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4801 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4861 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4921 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4981 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     5041 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     5101 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5161 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5221 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5281 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     5341 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5401 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5461 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     5521 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5581 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     5641 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5701 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5761 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5821 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5881 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5941 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6001 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6061 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6121 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6181 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6241 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6301 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6361 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6421 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6481 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     6541 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6601 cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6661 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6721 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6781 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6841 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6901 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6961 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7021 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7081 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7141 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7201 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7261 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7321 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7381 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7441 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7501 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7561 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7621 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7681 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7741 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7801 atttccccga aaagtgccac ctgacgtc
//
