LOCUS       V043961                 7409 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043961
VERSION     V043961
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             951..2225
                     /label="RBBP4(NM_005610)"
                     /note="RBBP4(NM_005610)"
                     /gene="RBBP4"
     CDS             2247..2942
                     /codon_start=1
                     /note=""
                     /product="promiscuous R40G mutant of a biotin protein
                     ligase from Aquifex aeolicus (Kim et al., 2016)"
                     /transl_table=1
                     /translation="FKNLIWLKEVDSTQERLKEWNVSYGTALVADRQTKGRGGLGRKWL
                     SQEGGLYFSFLLNPKEFENLLQLPLVLGLSVSEALEEITEIPFSLKWPNDVYFQEKKVS
                     GVLCELSKDKLIVGIGINVNQREIPEEIKDRATTLYEITGKDWDRKEVLLKVLKRISEN
                     LKKFKEKSFKEFKGKIESKMLYLGEEVKLLGEGKITGKLVGLSEKGGALILTEEGIKEI
                     LSGEFSLRRS"
                     /label="BioID2"
     CDS             2943..2969
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    3005..3229
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3275..3703
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3717..4047
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4114..4908
                     /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    5082..5203
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5252..5268)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5276..5292
                     /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(5300..5330)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5345..5366
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5654..6242)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6413..7273)
                     /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(7274..7378)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttaaggcc tgttaaccgg tcgtacgtct ccggattcga attcgccacc atggccgaca
      961 aggaagcagc cttcgacgac gcagtggaag aacgagtgat caacgaggaa tacaaaatat
     1021 ggaaaaagaa cacccctttt ctttatgatt tggtgatgac ccatgctctg gagtggccca
     1081 gcctaactgc ccagtggctt ccagatgtaa ccagaccaga agggaaagat ttcagcattc
     1141 atcgacttgt cctggggaca cacacatcgg atgaacaaaa ccatcttgtt atagccagtg
     1201 tgcagctccc taatgatgat gctcagtttg atgcgtcaca ctacgacagt gagaaaggag
     1261 aatttggagg ttttggttca gttagtggaa aaattgaaat agaaatcaag atcaaccatg
     1321 aaggagaagt aaacagggcc cgttatatgc cccagaaccc ttgtatcatc gcaacaaaga
     1381 ctccttccag tgatgttctt gtttttgact atacaaaaca tccttctaaa ccagatcctt
     1441 ctggagagtg caacccagac ttgcgtctcc gtggacatca gaaggaaggc tatgggcttt
     1501 cttggaaccc aaatctcagt gggcacttac ttagtgcttc agatgaccat accatctgcc
     1561 tgtgggacat cagtgccgtt ccaaaggagg gaaaagtggt agatgcgaag accatcttta
     1621 cagggcatac ggcagtagta gaagatgttt cctggcatct actccatgag tctctgtttg
     1681 ggtcagttgc tgatgatcag aaacttatga tttgggatac tcgttcaaac aatacttcca
     1741 aaccaagcca ctcagttgat gctcacactg ctgaagtgaa ctgcctttct ttcaatcctt
     1801 atagtgagtt cattcttgcc acaggatcag ctgacaagac tgttgccttg tgggatctga
     1861 gaaatctgaa acttaagttg cattcctttg agtcacataa ggatgaaata ttccaggttc
     1921 agtggtcacc tcacaatgag actattttag cttccagtgg tactgatcgc agactgaatg
     1981 tctgggattt aagtaaaatt ggagaggaac aatccccaga agatgcagaa gacgggccac
     2041 cagagttgtt gtttattcat ggtggtcata ctgccaagat atctgatttc tcctggaatc
     2101 ccaatgaacc ttgggtgatt tgttctgtat cagaagacaa tatcatgcaa gtgtggcaaa
     2161 tggcagagaa catttataat gatgaagacc ctgaaggaag cgtggatcca gaaggacaag
     2221 ggtccggtgg aggcggatca ggatccttca agaacctgat ctggctgaag gaggtggaca
     2281 gcacccagga gagactgaag gagtggaacg tgagctacgg caccgccctg gtggccgaca
     2341 gacagaccaa gggcagaggc ggcctgggca gaaagtggct gagccaggag ggcggcctgt
     2401 acttcagctt cctgctgaac cccaaggagt tcgagaacct gctgcagctg cccctggtgc
     2461 tgggcctgag cgtgagcgag gccctggagg agatcaccga gatccccttc agcctgaagt
     2521 ggcccaacga cgtgtacttc caggagaaga aggtgagcgg cgtgctgtgc gagctgagca
     2581 aggacaagct gatcgtgggc atcggcatca acgtgaacca gagagagatc cccgaggaga
     2641 tcaaggacag agccaccacc ctgtacgaga tcaccggcaa ggactgggac agaaaggagg
     2701 tgctgctgaa ggtgctgaag agaatcagcg agaacctgaa gaagttcaag gagaagagct
     2761 tcaaggagtt caagggcaag atcgagagca agatgctgta cctgggcgag gaggtgaagc
     2821 tgctgggcga gggcaagatc accggcaagc tggtgggcct gagcgagaag ggcggcgccc
     2881 tgatcctgac cgaggagggc atcaaggaga tcctgagcgg cgagttcagc ctgagaagaa
     2941 gctaccctta cgatgtaccg gattacgcat agaagcttaa gtttaaaccg ctgatcagcc
     3001 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     3061 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     3121 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     3181 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     3241 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     3301 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3361 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3421 ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3481 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3541 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3601 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat
     3661 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     3721 gtcagttagg gtgtggaaag tccccaggct ccccaggcag gcagaagtat gcaaagcatg
     3781 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3841 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3901 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3961 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     4021 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     4081 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     4141 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4201 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4261 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     4321 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4381 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4441 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4501 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4561 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4621 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     4681 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4741 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4801 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4861 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     4921 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     4981 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     5041 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     5101 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     5161 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     5221 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     5281 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     5341 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5401 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5461 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5521 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5581 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5641 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5701 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5761 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5821 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     5881 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5941 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6001 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6061 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     6121 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6181 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6241 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6301 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6361 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6421 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6481 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6541 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6601 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6661 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6721 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6781 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6841 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6901 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6961 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7021 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7081 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7141 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7201 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7261 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7321 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7381 catttccccg aaaagtgcca cctgacgtc
//
