LOCUS       V046105                 8221 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046105
VERSION     V046105
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             931..3657
                     /label="Usp33(NM_133247)"
                     /note="Usp33(NM_133247)"
                     /gene="Usp33"
     CDS             3664..3753
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     polyA_signal    3788..4012
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4058..4486
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4500..4829
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4896..5690
                     /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    5864..5985
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6034..6050)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6058..6074
                     /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(6082..6112)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6127..6148
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6436..7021)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7192..8052)
                     /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(8053..8157)
                     /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 cactagtcca gtgtggtgga attcgccacc atgacgactt ttcgaaatca ttgtccacat
      961 ttggattcag ttggtgaaat aacgaaagag gacttgatac agaaatctct cggcgcttgt
     1021 caggactgta aagtcagagg accaaacctg tgggcgtgcc tggagaatag gtgctcctat
     1081 gttggctgcg gggagtcgca agtggaccac agcaccatac actcgcagga gacaaagcat
     1141 tatctaactg tgaacctcac cactcttcga gtatggtgtt atgcttgcag caaagaagta
     1201 tttttggata gaaaattagg aactcctcct tcattacctc atgtaagaca gcctcaacaa
     1261 acacaagaaa acagtgtcca ggattttaaa attcccagta atccagcatt gaaaaccccc
     1321 atggttgctg tgtctgaaga tctggatata gaagtggaag aggaagacga gctgaaggct
     1381 agaggcctga caggtttgaa aaacattgga aatacttgtt atatgaacgc ggcgctgcag
     1441 gctctttcta actgcccacc tttgacacag ttctttcttg attgtggagg actggctaga
     1501 acagataaga aaccagcaat ttgtaaaagt tatctcaaac taatgaccga gttgtggcac
     1561 aagagcaggc caggatctgt tgtgccggct aatctgttcc aaggaattaa aactgtaaat
     1621 ccaacttttc gaggttattc tcagcaggat gcccaggaat tccttcgctg tctaatggac
     1681 ctgcttcatg aggagctgaa ggagcaggtc atggaaatgg aggaagagcc tcaaacacta
     1741 acttctgagg agacggtgga ggaagagaag agccagtcag atgtggattt tcagtcgtgc
     1801 gagtcttgta gcagcagcga gaaagcagaa aatgagagtg gctccaaagg ctttcctgag
     1861 gacagcaatg agaccaccat gctcatccag gacgaggatg acctggagat ggccaaagac
     1921 tggcagaaag agaaggtgtg caataagatc aacaaggcaa atgccgatgt agaactggac
     1981 aaagacaggg acacagtgtg tgaaacagtt gacctaaaca gccaggagac cgtcaaagtg
     2041 cagatacacg gcagagcctc agaatctatc actgatgtcc atctgaatga cctagctacg
     2101 tcacagatcc ttccttcaaa tgaaagtgtt agtccacggt tatcagcaag ccctcctaag
     2161 ctaggcagtc tgtggccagg actgtcgcct ccacacaaga aagctcagtc tacatctgca
     2221 aagaggaaaa agcagcataa gaaatacaga agtgtcatct ccgacatatt cgatggaaca
     2281 gtcattagct cggtacagtg tctgacgtgc gatagggtgt ctataaccct cgagaccttt
     2341 caggatctgt ccttgccgat tcctggcaag gaggacctgg ctaagctgca ctcctccagt
     2401 cacccaacta tagtcaaagc agggtcatgt ggtgaagcgt acgccccgca ggggtggata
     2461 gctttcttca tggagtatgt gaagaggttt gttgtctcat gtgtccctag ctggttttgg
     2521 ggtccagtag ttaccttgca agattgtctt gctgccttct tcgccagaga tgaacttaaa
     2581 ggtgacaaca tgtacagttg tgaaaaatgc aaaaagttga ggaatggagt aaagttttgt
     2641 aaagtacaga agtttcctga gattttgtgt atccacctta aaagatttcg acatgaactg
     2701 atgttttcca ccaaaattag cacccatgtt tccttccccc tggaaggcct cgatcttcag
     2761 ccatttcttg caaaggacag cccagctcag attgtgacat atgatctcct gtcagtcatc
     2821 tgtcaccatg ggactgcaag tagtgggcac tacattgcct actgccgaaa caatttaaat
     2881 aacctgtggt atgagtttga tgaccagagc gtcactgaag tttcagagtc cactgtacag
     2941 aatgccgagg cctacgtcct tttctacagg aagagcagtg aagaggcaca aaaggagagg
     3001 cggaggatat caaatttgtt gaacatcatg gaacctagcc tccttcagtt ctacatatct
     3061 cgacagtggt tgaataaatt taagaccttt gctgaacctg gccctatttc aaataatgat
     3121 tttctctgta tccatggagg tattcctcca cgaaaagcga gttatattga agacttagtt
     3181 ttgatgctgc ctcagaacat ttgggataac ctctatagca ggtatggagg agggcctgct
     3241 gtcaaccatc tctacatctg ccacacctgc caaattgagt tagagaagat tgaaaaacga
     3301 agaaaaaccg aattggaaat ttttattcgg ctcaacagag catttcaaga ggaggactcc
     3361 ccagctactt tttactgtat cagcatgcag tggtttagag aatgggagag ttttgtaaag
     3421 ggtaaggatg gagatccccc aggtccaatc gacaacacta aaattgcggt tactaaatgt
     3481 ggcagtgtga tgctcaagca aggagcagac tctggtcaaa tttcagaaga aacatggaat
     3541 ttcctgcagt ctatatatgg tggggggcct gaagttatcc tccgacctcc agttgttcat
     3601 gttgaccctg atgtactcca agcagaggaa aagattgaag tagaaactcg ctctttgctc
     3661 gaggaacaaa aactaatatc agaggaagat ttggaacaga aactaatcag tgaagaagat
     3721 ttagagcaaa agttaatttc cgaggaggac ttatgagggc ccgtttaaac ccgctgatca
     3781 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     3841 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     3901 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     3961 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     4021 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     4081 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4141 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4201 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4261 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4321 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4381 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     4441 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     4501 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4561 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4621 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4681 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4741 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     4801 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     4861 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     4921 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4981 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     5041 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     5101 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     5161 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     5221 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     5281 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     5341 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     5401 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     5461 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5521 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5581 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5641 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     5701 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5761 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5821 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     5881 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5941 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     6001 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     6061 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     6121 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     6181 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     6241 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     6301 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6361 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6421 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6481 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6541 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6601 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6661 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6721 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6781 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6841 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     6901 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6961 cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7021 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7081 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7141 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7201 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7261 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7321 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7381 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7441 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7501 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7561 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7621 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7681 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7741 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7801 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7861 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7921 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7981 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     8041 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8101 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8161 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     8221 t
//
