LOCUS       V040958                 8128 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040958
VERSION     V040958
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"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             991..3645
                     /label="KMT5B(NM_017635)"
                     /note="KMT5B(NM_017635)"
                     /gene="KMT5B"
     polyA_signal    3725..3949
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3995..4423
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4437..4766
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4833..5627
                     /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    5801..5922
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5971..5987)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5995..6011
                     /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(6019..6049)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6064..6085
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6373..6961)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7132..7992)
                     /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(7993..8097)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 gtttaaactt aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt atgaagtggt tgggagaatc caagaacatg
     1021 gtggtgaatg gcaggagaaa tggaggcaag ttgtctaatg accatcagca gaatcaatca
     1081 aaattacagc acacggggaa ggacaccctg aaggctggca aaaatgcagt cgagaggagg
     1141 tcgaacagat gtaatggtaa ctcgggattt gaaggacaga gtcgctatgt accatcctct
     1201 ggaatgtccg ccaaggaact ctgtgaaaat gatgacctag caaccagttt ggttcttgat
     1261 ccctatttag gttttcaaac acacaaaatg aatactagcg cctttccttc gaggagctca
     1321 aggcattttt caaaatctga cagtttttct cacaacaacc ctgtgagatt taggcctatt
     1381 aaaggaaggc aggaagaact aaaggaagta attgaacgtt ttaagaaaga tgaacacttg
     1441 gagaaagcct tcaaatgttt gacttcaggc gaatgggcac ggcactattt tctcaacaag
     1501 aataaaatgc aggagaaatt attcaaagaa catgtattta tttatttgcg aatgtttgca
     1561 actgacagtg gatttgaaat attgccatgt aatagatact catcagaaca aaatggagcc
     1621 aaaatagttg caacaaaaga gtggaaacga aatgacaaaa tagaattact ggtgggttgt
     1681 attgccgaac tttcagaaat tgaggagaac atgctactta gacatggaga aaacgacttc
     1741 agtgtcatgt actccacaag gaaaaactgt gctcaactct ggctgggtcc tgctgcgttt
     1801 ataaaccatg attgcagacc taattgtaag tttgtgtcaa ctggtcgaga tacagcatgt
     1861 gtgaaggctc taagagacat tgaacctgga gaagaaattt cttgttatta tggagatggg
     1921 ttctttggag aaaataatga gttctgcgag tgttacactt gcgaaagacg gggcactggt
     1981 gcttttaaat ccagagtggg actgcctgcg cctgctcctg ttatcaatag caaatatgga
     2041 ctcagagaaa cagataaacg tttaaatagg cttaaaaagt taggtgacag cagcaaaaat
     2101 tcagacagtc aatctgtcag ctctaacact gatgcagata ccactcagga aaaaaacaat
     2161 gcaacttcta accgaaaatc ttcagttggc gtaaaaaaga atagcaagag cagaacgtta
     2221 acgaggcaat ctatgtcaag aattccagct tcttccaact ctacctcatc taagctaact
     2281 catataaata attccagggt accaaagaaa ctgaagaagc ctgcaaagcc tttactttca
     2341 aagataaaat tgagaaatca ttgcaagcgg ctggagcaaa agaatgcttc aagaaaactc
     2401 gaaatgggaa acttagtact gaaagagcct aaagtagttc tgtataaaaa tttgcccatt
     2461 aaaaaagata aggagccaga gggaccagcc caagccgcag ttgccagcgg gtgcttgact
     2521 agacacgcgg cgagagaaca cagacagaat cctgtgagag gtgctcattc gcagggggag
     2581 agctcgccct gcacctacat aactcggcgg tcagtgagga caagaacaaa tctgaaggag
     2641 gcctctgaca tcaagcttga accaaatacg ttgaatggct ataaaagcag tgtgacggaa
     2701 ccttgccccg acagtggtga acagctgcag ccagctcctg tgctgcagga ggaagaactg
     2761 gctcatgaga ctgcacaaaa aggggaggca aagtgtcata agagtgacac aggcatgtcc
     2821 aaaaagaagt cacgacaagg aaaacttgtg aaacagtttg caaaaataga ggaatctact
     2881 ccagtgcacg attctcctgg aaaagacgac gcggtaccag atttgatggg tccccattct
     2941 gaccagggtg agcacagtgg cactgtgggc gtgcctgtga gctacacaga ctgtgctcct
     3001 tcacccgtcg gttgttcagt tgtgacatca gatagcttca aaacaaaaga cagctttaga
     3061 actgcaaaaa gtaaaaagaa gaggcgaatc acaaggtatg atgcacagtt aatcctagaa
     3121 aataactctg ggattcccaa attgactctt cgtaggcgtc atgatagcag cagcaaaaca
     3181 aatgaccaag agaatgatgg aatgaactct tccaaaataa gcatcaagtt aagcaaagac
     3241 catgacaacg ataacaatct ctatgtagca aagcttaata atggatttaa ctcaggatca
     3301 ggcagtagtt ctacaaaatt aaaaatccag ctaaaacgag atgaggaaaa tagggggtct
     3361 tatacagagg ggcttcatga aaatggggtg tgctgcagtg atcctctttc tctcttggag
     3421 tctcgaatgg aggtggatga ctatagtcag tatgaggaag aaagtacaga tgattcctcc
     3481 tcttctgagg gcgatgaaga ggaggatgac tatgatgatg actttgaaga cgattttatt
     3541 cctcttcctc cagctaagcg cttgaggtta atagttggaa aagactctat agatattgac
     3601 atttcttcaa ggagaagaga agatcagtct ttaaggctta atgcctaaga attctgcaga
     3661 tatccagcac agtggcggcc gctcgagtct agagggcccg tttaaacccg ctgatcagcc
     3721 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     3781 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     3841 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     3901 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     3961 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     4021 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4081 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4141 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4201 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4261 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4321 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     4381 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     4441 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4501 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4561 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4621 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4681 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     4741 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     4801 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4861 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4921 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4981 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     5041 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5101 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5161 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5221 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5281 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5341 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5401 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5461 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5521 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5581 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5641 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5701 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5761 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     5821 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5881 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     5941 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6001 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6061 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6121 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6181 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6241 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6301 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6361 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6421 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6481 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6541 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6601 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6661 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6721 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6781 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     6841 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6901 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6961 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7021 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7081 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7141 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7201 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7261 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7321 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7381 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7441 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7501 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7561 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7621 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7681 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7741 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7801 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7861 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7921 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7981 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8041 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8101 atttccccga aaagtgccac ctgacgtc
//
