LOCUS       V027780                 8373 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027780
VERSION     V027780
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             706..3099
                     /label="USP10(NM_005153)"
                     /note="USP10(NM_005153)"
                     /gene="USP10"
     CDS             3106..3129
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHHHH"
                     /label="8xHis"
     misc_feature    3149..3737
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3803..3851
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4053..4481
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4495..4824
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4891..5685
                     /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    5861..5982
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6031..6047)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6055..6071
                     /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(6079..6109)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6124..6145
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6433..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 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatggc cctccacagc
      721 ccgcagtata tttttggaga ttttagccct gatgaattca atcaattctt tgtgactcct
      781 cgatcttcag ttgagcttcc tccatacagt ggaacagttc tgtgtggcac acaggctgtg
      841 gataaactac ctgatggaca agaatatcag agaattgagt ttggtgtcga tgaagtcatt
      901 gaacccagtg acactttgcc gagaaccccc agctacagta tttcaagcac actgaaccct
      961 caggcccctg aatttattct cggttgtaca gcttccaaaa taacccctga tggtatcact
     1021 aaagaagcaa gctatggctc catcgactgc cagtacccag gctctgccct cgctttggat
     1081 ggaagttcta atgtggaggc ggaagttttg gaaaatgatg gtgtctcagg tggtcttgga
     1141 caaagggagc gtaaaaagaa gaaaaagcgg ccacctggat attacagcta tttgaaagat
     1201 ggtggcgatg atagtatctc cacagaagcc ctggtcaatg gccatgccaa ttcagcagtc
     1261 ccgaacagtg tcagtgcaga ggatgcagaa tttatgggtg acatgccccc gtcagttacg
     1321 cccaggactt gtaacagccc ccagaactcc acagactctg tcagtgacat tgtgcctgac
     1381 agtcctttcc ccggagcact cggcagtgac accaggactg cagggcagcc agaggggggc
     1441 cccggggctg attttggtca gtcctgcttc cctgcagagg ctggcagaga caccctgtca
     1501 aggacagctg gggctcagcc ctgcgttggt accgatacta ctgaaaacct tggagttgct
     1561 aatggacaaa tacttgaatc ctcgggtgag ggcacagcta ccaacggggt ggagttgcac
     1621 accacggaaa gcatagactt ggacccaacc aaacccgaga gtgcatcacc tcctgctgac
     1681 ggcacgggct ctgcatcagg cacccttcct gtcagccagc ccaagtcctg ggccagcctc
     1741 tttcatgatt ctaagccctc ttcctcctcg ccggtggcct atgtggaaac taagtattcc
     1801 cctcccgcca tatctcccct ggtttctgaa aagcaggttg aagtcaaaga agggcttgtt
     1861 ccggtttcag aggatcctgt agccataaag attgcagagt tgctggagaa tgtaacccta
     1921 atccataaac cagtgtcgtt gcaaccccgt gggctgatca ataaagggaa ctggtgctac
     1981 attaatgcta cactgcaggc attggttgct tgcccgccga tgtaccacct gatgaagttc
     2041 attcctctgt attccaaagt gcaaaggcct tgtacgtcaa cacccatgat agacagcttt
     2101 gttcggctaa tgaatgagtt cactaatatg ccagtacctc caaaaccccg acaagctctt
     2161 ggagataaaa tcgtgaggga tattcgccct ggagctgcct ttgagcccac atatatttac
     2221 agactcctga cagttaacaa gtcaagcctg tctgaaaagg gtcgacaaga agatgctgag
     2281 gaatacttag gcttcattct aaatggactt catgaggaaa tgttgaacct aaagaagctt
     2341 ctctcaccaa gtaatgaaaa acttacgatt tccaacggcc ccaaaaacca ctcggtcaat
     2401 gaagaagagc aggaagaaca aggtgaagga agcgaggatg aatgggaaca agtgggcccc
     2461 cggaacaaga cttccgtcac ccgccaggcg gattttgttc agactccaat caccggcatt
     2521 tttggtggac acatcaggtc tgtggtttac cagcagagtt caaaagaatc tgccactttg
     2581 cagccatttt tcacgttgca gttggatatc cagtcagaca agatacgcac agtccaggat
     2641 gcactggaga gcttggtggc aagagaatct gtccaaggtt ataccacaaa aaccaaacaa
     2701 gaggttgaga taagtcgaag agtgactctg gaaaaactcc ctcctgtcct cgtgctgcac
     2761 ctgaaacgat tcgtttatga gaagactggt gggtgccaga agcttatcaa aaatattgaa
     2821 tatcctgtgg acttggaaat tagtaaagaa ctgctttctc caggggttaa aaataagaat
     2881 tttaaatgcc accgaaccta tcggctcttt gcagtggtct accatcacgg caacagtgcg
     2941 acgggcggcc attacactac agacgtcttc cagatcggtc tgaatggctg gctgcgcatc
     3001 gatgaccaga cagtcaaggt gatcaaccag taccaggtgg tgaaaccaac tgctgaacgc
     3061 acagcctacc tcctgtatta ccgccgagtg gacctgctgc tcgagcacca tcaccatcac
     3121 catcaccatt agggcccgat atctcgacaa tcaacctctg gattacaaaa tttgtgaaag
     3181 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     3241 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     3301 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     3361 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     3421 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     3481 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     3541 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     3601 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     3661 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     3721 ttgggccgcc tccccgcctg gaaacggggg aggctaactg aaacacggaa ggagacaata
     3781 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg
     3841 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     3901 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     3961 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga
     4021 tctgcgcagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     4081 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4141 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4201 aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4261 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4321 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4381 caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg
     4441 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     4501 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4561 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4621 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4681 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4741 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     4801 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     4861 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4921 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4981 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     5041 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     5101 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     5161 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     5221 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     5281 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     5341 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     5401 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     5461 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     5521 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     5581 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     5641 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     5701 tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5761 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5821 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     5881 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5941 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     6001 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6061 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6121 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6181 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6241 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6301 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6361 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6421 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6481 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6541 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6601 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     6661 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6721 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6781 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6841 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     6901 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6961 tggtagcggt 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 tcgactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     8281 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     8341 ttgaccgaca attgcatgaa gaatctgctt agg
//
