LOCUS       V023996                 8758 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023996
VERSION     V023996
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..4278
                     /label="KDM4B(NM_015015)"
                     /note="KDM4B(NM_015015)"
                     /gene="KDM4B"
     polyA_signal    4358..4582
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4628..5056
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5070..5399
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5466..6260
                     /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    6434..6555
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6604..6620)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6628..6644
                     /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(6652..6682)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6697..6718
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7006..7591)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7762..8622)
                     /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(8623..8727)
                     /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 atggggtctg aggaccacgg cgcccagaac
     1021 cccagctgta aaatcatgac gtttcgccca accatggaag aatttaaaga cttcaacaaa
     1081 tacgtggcct acatagagtc gcagggagcc caccgggcgg gcctggccaa gatcatcccc
     1141 ccgaaggagt ggaagccgcg gcagacgtat gatgacatcg acgacgtggt gatcccggcg
     1201 cccatccagc aggtggtgac gggccagtcg ggcctcttca cgcagtacaa tatccagaag
     1261 aaggccatga cagtgggcga gtaccgccgc ctggccaaca gcgagaagta ctgtaccccg
     1321 cggcaccagg actttgatga ccttgaacgc aaatactgga agaacctcac ctttgtctcc
     1381 ccgatctacg gggctgacat cagcggctct ttgtatgatg acgacgtggc ccagtggaac
     1441 atcgggagcc tccggaccat cctggacatg gtggagcgcg agtgcggcac catcatcgag
     1501 ggcgtgaaca cgccctacct gtacttcggc atgtggaaga ccaccttcgc ctggcacacc
     1561 gaggacatgg acctgtacag catcaactac ctgcactttg gggagcctaa gtcctggtac
     1621 gccatcccac cagagcacgg caagcgcctg gagcggctgg ccatcggctt cttccccggg
     1681 agctcgcagg gctgcgacgc cttcctgcgg cataagatga ccctcatctc gcccatcatc
     1741 ctgaagaagt acgggatccc cttcagccgg atcacgcagg aggccgggga attcatgatc
     1801 acatttccct acggctacca cgccggcttc aatcacgggt tcaactgcgc agaatctacc
     1861 aacttcgcca ccctgcggtg gattgactac ggcaaagtgg ccactcagtg cacgtgccgg
     1921 aaggacatgg tcaagatctc catggacgtg ttcgtgcgca tcctgcagcc cgagcgctac
     1981 gagctgtgga agcagggcaa ggacctcacg gtgctggacc acacgcggcc cacggcgctc
     2041 accagccccg agctgagctc ctggagtgca tcccgggcct cgctgaaggc caagctcctc
     2101 cgcaggtctc accggaaacg gagccagccc aagaagccga agcccgaaga ccccaagttc
     2161 cctggggagg gtacggctgg ggcagcgctc ctagaggagg ctgggggcag cgtgaaggag
     2221 gaggctgggc cggaggttga ccccgaggag gaggaggagg agccgcagcc actgccacac
     2281 ggccgggagg ccgagggcgc agaagaggac gggaggggca agctgcggcc aaccaaggcc
     2341 aagagcgagc ggaagaagaa gagcttcggc ctgctgcccc cacagctgcc gcccccgcct
     2401 gctcacttcc cctcagagga ggcgctgtgg ctgccatccc cactggagcc cccggtgctg
     2461 ggcccaggcc ctgcagccat ggaggagagc cccctgccgg caccccttaa tgtcgtgccc
     2521 cctgaggtgc ccagtgagga gctagaggcc aagcctcggc ccatcatccc catgctgtac
     2581 gtggtgccgc ggccgggcaa ggcagccttc aaccaggagc acgtgtcctg ccagcaggcc
     2641 tttgagcact ttgcccagaa gggtccgacc tggaaggaac cagtttcccc catggagctg
     2701 acggggccag aggacggtgc agccagcagt ggggcaggtc gcatggagac caaagcccgg
     2761 gccggagagg ggcaggcacc gtccacattt tccaaattga agatggagat caagaagagc
     2821 cggcgccatc ccctgggccg gccgcccacc cggtccccac tgtcggtggt gaagcaggag
     2881 gcctcaagtg acgaggaggc atcccctttc tccggggagg aagatgtgag tgacccggac
     2941 gccttgaggc cgctgctgtc tctgcagtgg aagaacaggg cggccagctt ccaggccgag
     3001 aggaagttca acgcagcggc tgcgcgcacg gagccctact gcgccatctg cacgctcttc
     3061 tacccctact gccaggccct acagactgag aaggaggcac ccatagcctc cctcggagag
     3121 ggctgcccgg ccacattacc ctccaaaagc cgtcagaaga cccgaccgct catccctgag
     3181 atgtgcttca cctctggcgg tgagaacacg gagccgctgc ctgccaactc ctacatcggc
     3241 gacgacggga ccagccccct gatcgcctgc ggcaagtgct gcctgcaggt ccatgccagt
     3301 tgctatggca tccgtcccga gctggtcaat gaaggctgga cgtgttcccg gtgcgcggcc
     3361 cacgcctgga ctgcggagtg ctgcctgtgc aacctgcgag gaggtgcgct gcagatgacc
     3421 accgatagga ggtggatcca cgtgatctgt gccatcgcag tccccgaggc gcgcttcctg
     3481 aacgtgattg agcgccaccc tgtggacatc agcgccatcc ccgagcagcg gtggaagctg
     3541 aaatgcgtgt actgccggaa gcggatgaag aaggtgtcag gtgcctgtat ccagtgctcc
     3601 tacgagcact gctccacgtc cttccacgtg acctgcgccc acgccgcagg cgtgctcatg
     3661 gagccggacg actggcccta tgtggtctcc atcacctgcc tcaagcacaa gtcggggggt
     3721 cacgctgtcc aactcctgag ggccgtgtcc ctaggccagg tggtcatcac caagaaccgc
     3781 aacgggctgt actaccgctg tcgcgtcatc ggtgccgcct cgcagacctg ctacgaagtg
     3841 aacttcgacg atggctccta cagcgacaac ctgtaccctg agagcatcac gagtagggac
     3901 tgtgtccagc tgggaccccc ttccgagggg gagctggtgg agctccggtg gactgacggc
     3961 aacctctaca aggccaagtt catctcctcc gtcaccagcc acatctacca ggtggagttt
     4021 gaggacgggt cccagctgac ggtgaagcgt ggggacatct tcaccctgga ggaggagctg
     4081 cccaagaggg tccgctctcg gctgtcactg agcacggggg caccgcagga gcccgccttc
     4141 tcgggggagg aggccaaggc cgccaagcgc ccgcgtgtgg gcaccccgct tgccacggag
     4201 gactccgggc ggagccagga ctacgtggcc ttcgtggaga gcctcctgca ggtgcagggc
     4261 cggcccggag cccccttcta ggaattctgc agatatccag cacagtggcg gccgctcgag
     4321 tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     4381 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     4441 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     4501 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     4561 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     4621 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4681 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4741 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4801 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4861 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4921 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4981 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     5041 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     5101 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     5161 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5221 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     5281 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     5341 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     5401 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     5461 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     5521 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     5581 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     5641 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     5701 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     5761 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     5821 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     5881 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     5941 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     6001 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     6061 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     6121 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     6181 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     6241 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     6301 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     6361 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     6421 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     6481 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     6541 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     6601 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     6661 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6721 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     6781 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     6841 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     6901 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     6961 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     7021 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     7081 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     7141 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     7201 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     7261 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     7321 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     7381 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     7441 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     7501 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg tttgcaagca
     7561 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7621 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7681 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7741 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7801 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7861 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7921 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     7981 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     8041 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     8101 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     8161 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     8221 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     8281 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     8341 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8401 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8461 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8521 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8581 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     8641 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     8701 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
