LOCUS       V045554                 8754 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045554
VERSION     V045554
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..3434
                     /label="Rbbp8(NM_001081223)"
                     /note="Rbbp8(NM_001081223)"
                     /gene="Rbbp8"
     CDS             3441..3506
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3530..4118
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4184..4232
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4434..4862
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4876..5205
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5272..6066
                     /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    6242..6363
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6412..6428)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6436..6452
                     /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(6460..6490)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6505..6526
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6814..7402)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7573..8433)
                     /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(8434..8538)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgag catttcagga agcggctgtg
      781 gaagccctaa ttctgcggat gcatccaatg acttcaagga actctggaca aaactaaaag
      841 agtatcatga taaggaagtg caaggtttac aagtcaaagt aaccaaactg aaaaaggaac
      901 gaattttaga tgcacagaga ctagaagaat tcttcaccaa aaatcaacag ctgagagacc
      961 agcagaaagt ccttcaggaa accattaaaa ttttagaaga taggttaaga gcaggattgt
     1021 gtgatcgctg tgcagtaact gaagagcata tgcacaaaaa gcagcaggag tttgaaaata
     1081 tccgacagca gaaccttaag cttattacag agctgatgaa tgaaaagaat actcttcaag
     1141 aagaaaacaa aaagctttct gaacaactgc agcagaaaat ggaaaatggt caacaggatc
     1201 aagtagctga gcttgcatgt gaggaaaaca ttattccaga ctcaccagta acatcctttt
     1261 ccttttctgg cattaaccgg ctacgaaaga aggagaatct ccatgtccga tacgtggaac
     1321 aaacacatac taagttggag cgctctctat gtacaaatga attaagaaaa atatctaagg
     1381 attcagctcc tgcaccagtt aactctgagg aacatgaaat tctcgtagct gacacttgtg
     1441 atcaaaatca ctcaccactg tcaaaaatat gtgaaacaag cagttacccc actgataaga
     1501 catcttttaa tttagacact gttgttgctg agacacttgg acttaatggt caagaagaat
     1561 ctgaacccca aggtcccatg agccctctgg gaagtgaact ctaccactgt ctcaaagaag
     1621 atcacaaaaa acaccctttt atggaatctg caagaagtaa ggaagatagc ttaagatttt
     1681 cagactctgc ttcaaagaca cctcctcaag aatttactac tcgggcgtca tctccggtat
     1741 ttggagctac ctctactgtg aaagctcatt tgggtttgaa tacaagtttc tccccctctc
     1801 tcttagacat tgggaaaaaa aaccttctga agacagctcc ctttagcaac attgctgttt
     1861 ctagatcaga gaaagttaga tccaaatctg aggataatgc ccttttcaca caacatagtc
     1921 ttgggtcaga agtgaaggtc attagccagt cattttctag taaacagata cttacaaata
     1981 aaactgtaag tgactctgta gatgaacagt gcagtgctga ccacatgaat actactgttg
     2041 ctgataaata tttggtgcct ttgaaatccc tgggaggcaa agcatccaaa aggaagagga
     2101 ctgaggagga aagtgagcat gcagtaaagt gcccccaggc ctgttttgat aaggaaaatg
     2161 cccttccttt cccaatggag aatcagttct ccatgaatgg agaccatgtg atggataaac
     2221 ctctggacct gtcagatcgg tttgcagcta ctcagcggca agagaagaac cacggaaacg
     2281 agacttctaa aaacaaactt aagcaagcca ctatttacga ggctttgaag cccattccaa
     2341 agggctcctc ctcaggccgt aaagccttga gtggggactg catgccagcc aaagactctt
     2401 gggagacata ctgcttacag ccgaggagcc tccagtcctc tagtaaattc tctccagacc
     2461 aaaaaacacc attacagata aaagaagaaa atcctgtctt caaaacacct ccatgttcac
     2521 aggaaagttt ggagacagag aatctctttg gtgatgtgaa gggtactggt tctcttgtgc
     2581 caacaaaagt aaagagcaga gcagtccatg gaggatgtga gcttgcatca gttcttcagt
     2641 taaacccctg tagagttgct aaaacaaagg ctctgccaag caaccaagat acgtcctttg
     2701 aaaatatcca gtggagtgta gacccaggag cagacctttc tcagtataaa atggatgtta
     2761 ctgtaataga tacaaaggat agcagtcact cacgattagg aggagaaaca gtggacatgg
     2821 actgtacatt ggtcagtgaa accgtgcttt taaaaatgaa gaaacaagag caaaaagaaa
     2881 gaagtccaaa tggagacata aaaatgaatg atagcctaga agatatgttt gatcggacaa
     2941 cacatgaaga gtatgagtcc tgtttggcag atagcttctc ccaggtacca gatgaggaag
     3001 aactgcctga caccacaaag aaaacaaaca tacctgctga taaacaagat ggagtcaaac
     3061 agaaagcatt tgtggggcca tattttaaag ataaggagag agaaactagc atacagaatt
     3121 ttcctcatat tgaggtagtt cggaaaaaag aagagagaag gaaattgctt gggcacacat
     3181 gtaaagaatg tgaaatttat tatgcagatt tgccagcaga agaaagagaa aagaagttgg
     3241 cttcctgctc aagacaccga tttcgctaca ttccacccaa cacaccagag aatttctggg
     3301 aagttggttt tccttctact cagacgtgtc tggaaagagg ttatatcaag gaagatctcg
     3361 atcttagtcc tcgtcccaaa agacggcagc cttacaacgc ggtcttttct ccaaaaggca
     3421 aagaacagag gaccggatcc gactacaaag accatgacgg tgattataaa gatcatgaca
     3481 tcgactacaa ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct
     3541 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     3601 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     3661 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     3721 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     3781 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3841 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3901 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     3961 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     4021 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     4081 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact
     4141 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     4201 aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     4261 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     4321 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     4381 ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc
     4441 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4501 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4561 cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt
     4621 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4681 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4741 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4801 tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4861 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4921 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4981 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     5041 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     5101 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     5161 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     5221 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     5281 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     5341 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     5401 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     5461 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     5521 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     5581 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     5641 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     5701 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5761 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     5821 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5881 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5941 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     6001 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     6061 ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc
     6121 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     6181 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     6241 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     6301 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     6361 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     6421 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     6481 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     6541 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6601 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6661 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6721 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6781 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6841 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6901 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6961 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     7021 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7081 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7141 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7201 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     7261 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7321 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7381 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7441 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7501 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7561 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7621 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7681 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7741 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7801 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7861 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7921 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7981 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     8041 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8101 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8161 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8221 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8281 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8341 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8401 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8461 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8521 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     8581 agatctcccg atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta
     8641 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     8701 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
