LOCUS       V027752                 7471 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027752
VERSION     V027752
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..2130
                     /label="TRIM21(NM_003141)"
                     /note="TRIM21(NM_003141)"
                     /gene="TRIM21"
     CDS             2137..2163
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2167..2193
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2197..2223
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    2247..2835
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2901..2949
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3151..3579
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3593..3922
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3989..4783
                     /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    4959..5080
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5129..5145)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5153..5169
                     /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(5177..5207)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5222..5243
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5531..6119)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6290..7150)
                     /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(7151..7255)
                     /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 ttcagcagca
      721 cgcttgacaa tgatgtggga ggaggtcaca tgccctatct gcctggaccc cttcgtggag
      781 cctgtgagca tcgagtgtgg ccacagcttc tgccaggaat gcatctctca ggttgggaaa
      841 ggtgggggca gcgtctgtcc tgtgtgccgg cagcgctttc tgctcaagaa tctccggccc
      901 aatcgacagc tagccaacat ggtgaacaac cttaaagaaa tcagccagga ggccagagag
      961 ggcacacagg gggaacggtg tgcagtgcat ggagagagac ttcacctgtt ctgtgagaaa
     1021 gatgggaagg ccctttgctg ggtatgtgcc cagtctcgga aacaccgtga ccacgccatg
     1081 gtccctcttg aggaggctgc acaggagtac caggagaagc tccaggtggc attaggggaa
     1141 ctgagaagaa agcaggagtt ggctgagaag ttggaagtgg aaattgcaat aaagagagca
     1201 gactggaaga aaacagtgga aacacagaaa tctaggattc acgcagagtt tgtgcagcaa
     1261 aaaaacttcc tggttgaaga agaacagagg cagctgcagg agctggagaa ggatgagagg
     1321 gagcagctga gaatcctggg ggagaaagag gccaagctgg cccagcagag ccaggcccta
     1381 caggagctca tctcagagct agatcgaagg tgccacagct cagcactgga actgctgcag
     1441 gaggtgataa ttgtcctgga aaggagtgag tcctggaacc tgaaggacct ggatattacc
     1501 tctccagaac tcaggagtgt gtgccatgtg ccagggctga agaagatgct gaggacatgt
     1561 gcagtccaca tcactctgga tccagacaca gccaatccgt ggctgatact ttcagaagat
     1621 cggagacaag tgaggcttgg agacacccag cagagcatac ctggaaatga agagagattt
     1681 gatagttatc ctatggtcct gggtgcccag cactttcact ctggaaaaca ttactgggag
     1741 gtagatgtga caggaaagga ggcctgggac ctgggtgtct gcagagactc tgtgcgcagg
     1801 aaggggcact ttttgcttag ttccaagagt ggcttctgga caatttggtt gtggaacaaa
     1861 caaaaatatg aggctggcac ctacccccag actcccctcc accttcaggt gcctccatgc
     1921 caagttggga ttttcctgga ctatgaggct ggcatggtct ccttctacaa catcactgac
     1981 catggctccc tcatctactc cttctctgaa tgtgccttta caggacctct gcggcccttc
     2041 ttcagtcctg gtttcaatga tggaggaaaa aacacagccc ctctaaccct ctgtccactg
     2101 aatattggat cacaaggatc cactgactat ctcgagtacc catacgacgt acctgattat
     2161 gcaacttatc cttatgatgt acctgattat gctagctatc cttatgatgt cccagactac
     2221 gcatagtgag ggcccgatat ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat
     2281 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     2341 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     2401 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     2461 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     2521 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     2581 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     2641 agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     2701 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     2761 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     2821 gggccgcctc cccgcctgga aacgggggag gctaactgaa acacggaagg agacaatacc
     2881 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt
     2941 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     3001 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     3061 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcagatc
     3121 tgcgcagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     3181 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3241 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3301 atcggggcat ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3361 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3421 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3481 accctatctc ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt
     3541 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     3601 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3661 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3721 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3781 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3841 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     3901 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     3961 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     4021 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4081 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4141 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     4201 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4261 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4321 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4381 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4441 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4501 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4561 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4621 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4681 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4741 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4801 gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4861 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4921 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     4981 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5041 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     5101 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5161 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5221 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5281 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5341 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5401 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5461 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5521 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5581 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5641 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5701 cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg
     5761 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5821 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5881 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5941 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     6001 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6061 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6121 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6181 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6241 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6301 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6361 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6421 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6481 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6541 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6601 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6661 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6721 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6781 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6841 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6901 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6961 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7021 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7081 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7141 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7201 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7261 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtc
     7321 gactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
     7381 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
     7441 gaccgacaat tgcatgaaga atctgcttag g
//
