LOCUS       V018319                 9487 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018319
VERSION     V018319
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             1033..5040
                     /label="KDM2B(NM_032590)"
                     /note="KDM2B(NM_032590)"
                     /gene="KDM2B"
     polyA_signal    5087..5311
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5357..5785
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5799..6128
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6195..6989
                     /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    7163..7284
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7333..7349)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7357..7373
                     /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(7381..7411)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7426..7447
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7735..8320)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8491..9351)
                     /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(9352..9456)
                     /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 ggtaccgagc tcggatccac tagtccagtg
     1021 tggtggaatt cgatggcggg tccgcaaatg gggggatctg cagaggatca ccccccacga
     1081 aaaagacatg cagcagaaaa gcaaaaaaag aaaacagtta tatatacaaa atgctttgaa
     1141 tttgagtcgg ccacacagcg cccgattgac cgccagcgat acgacgagaa cgaggacttg
     1201 tcggacgtgg aggagatcgt cagcgtccgc ggcttcagcc tggaggagaa gcttcgcagc
     1261 cagctgtacc agggggactt cgtgcacgcc atggagggca aagatttcaa ctatgagtac
     1321 gtacagagag aagctctcag ggttcccctg atatttcgag aaaaggatgg actgggaatt
     1381 aagatgcctg accctgattt cacagtccga gacgtcaaac tcctagtggg gagccggcgg
     1441 cttgtggacg tgatggatgt gaacacccag aagggcacgg agatgagcat gtcccagttt
     1501 gtgcgttact acgagacgcc cgaggcccag cgggacaagc tgtacaacgt catcagccta
     1561 gagttcagcc acaccaagct ggagcacttg gtcaagcgtc cgactgtggt agacctggtg
     1621 gactgggtgg acaacatgtg gccccagcat ctgaaggaga agcagacaga agccacgaac
     1681 gccattgcag agatgaagta cccgaaagtg aaaaagtact gtctgatgag cgtgaaaggt
     1741 tgtttcaccg acttccacat cgactttgga ggcacttccg tttggtacca tgttttccgg
     1801 ggtgggaaga ttttttggct gattcctcca acgctgcaca atttggcgct gtacgaggag
     1861 tgggtgctgt caggcaaaca gagtgacatc tttctgggag accgtgtgga acgatgccaa
     1921 agaattgagc tgaagcaggg ctacacattt ttcatccctt ccggttggat ccatgccgtc
     1981 tacacccctg tagactcttt ggtgttcggc ggaaacatcc tgcacagctt taacgtgccc
     2041 atgcagctgc ggatctacga gatcgaggac aggacgcggg tgcagcccaa attccgttac
     2101 cccttctact atgagatgtg ctggtatgtc ctggagagat acgtgtactg tgtgacccag
     2161 cgctcccacc tcactcagga ataccagagg gagtcgatgc ttattgatgc cccgaggaag
     2221 cccagcatag acggcttctc ttcggattcc tggctggaga tggaggagga ggcctgtgat
     2281 cagcagcctc aggaggagga ggagaaggac gaggagggcg agggcaggga cagggcaccc
     2341 aaaccgccca ccgatggctc cacttcaccc accagcacgc cctctgagga ccaggaggcc
     2401 ctcgggaaga agcccaaagc acctgccctg cgattcctca aaaggacttt gtctaatgag
     2461 tcggaggaaa gtgtgaagtc caccacattg gccgtagact accccaagac ccccaccggc
     2521 tctcccgcca cggaggtctc tgccaaatgg acccatctca ctgagtttga actgaagggc
     2581 ctgaaagctc tggtggagaa actggaatcc ctcccggaga acaagaagtg tgtccccgag
     2641 ggcatcgagg acccccaggc actcctggag ggtgtgaaga acgtcctgaa ggagcacgca
     2701 gatgatgacc ctagtctggc catcactggg gtccctgtgg tgacttggcc aaagaagact
     2761 ccaaagaacc gggctgtggg tcggcccaag gggaagctgg gcccggcctc cgcggtgaag
     2821 ttggccgcca accggacaac ggcaggagct cggcggcgcc ggacgcgatg ccgcaagtgc
     2881 gaggcctgcc tgcggaccga gtgcggagag tgccacttct gcaaggacat gaagaagttc
     2941 gggggccccg ggcgcatgaa gcagagctgc atcatgcggc agtgcatcgc gccagtgctg
     3001 ccccacaccg ccgtgtgcct tgtgtgtggc gaggcgggga aggaagacac ggtggaagag
     3061 gaggaaggca agtttaacct catgctcatg gagtgctcca tctgcaatga aatcatccac
     3121 cctggatgcc ttaagattaa ggagtcagag ggtgtggtca acgacgagct tccaaactgc
     3181 tgggagtgtc cgaagtgtaa ccacgccggc aagaccggga aacaaaagcg tggccctggc
     3241 tttaagtacg cctccaacct gcccggctcc ctgctcaagg agcagaagat gaaccgggac
     3301 aacaaggaag ggcaggaacc tgccaagcgg aggagtgagt gtgaggaggc gccccggcgc
     3361 aggtcggatg agcactcgaa gaaggtgccg ccggacggcc ttctgcgcag aaagtctgac
     3421 gacgtgcacc tgaggaagaa gcggaaatac gagaagcccc aggagctgag tggacgcaag
     3481 cgggcctcat cgcttcaaac gtcccccggt tcctcctctc acctctcgcc gaggccccct
     3541 ctaggcagca gcctcagccc ctggtggaga tccagtctca cttacttcca gcagcagctc
     3601 aaacctggca aagaagataa gcttttcagg aaaaagcggc ggtcctggaa gaacgccgag
     3661 gaccgcatgg cgctggccaa caagcccctc cggcgcttca agcaggaacc cgaggacgaa
     3721 ctgcccgagg cgccccccaa gaccagggag agcgaccact cccgctccag ctcccccacc
     3781 gcgggaccca gcaccgaagg ggccgagggc ccggaggaga agaagaaggt gaagatgcgc
     3841 cggaagcggc ggcttcccaa caaggagctg agcagggagc tgagcaagga gctcaaccac
     3901 gagatccaga ggacggagaa cagcctggcc aacgagaacc agcagcccat caagtcggag
     3961 cctgagagcg agggcgagga gcccaagcgg cccccgggca tctgcgagcg tccccaccgc
     4021 ttcagcaagg ggctcaacgg caccccccgg gagctgcggc accagctggg gcccagcctg
     4081 cgcagcccgc cccgtgtcat ctcccggccc ccaccctccg tgtccccgcc caagtgtatc
     4141 cagatggagc gccatgtgat ccggccaccc cccatcagcc ccccgcctga ctcgctaccc
     4201 ctggacgatg gggcagccca cgtcatgcac agggaggtgt ggatggccgt cttcagctac
     4261 ctcagccacc aagacctgtg tgtgtgcatg cgggtctgca ggacctggaa ccgctggtgc
     4321 tgcgataagc ggttgtggac ccgcattgac ctgaaccact gcaagtctat cacacccctg
     4381 atgctgagtg gcatcatccg gcgacagccc gtctccctcg acctcagctg gaccaatatc
     4441 tccaagaagc agctgagctg gctcatcaac cggctgcctg ggctccggga cttggtgctg
     4501 tcaggctgct catggatcgc ggtctcggcc ctttgcagct ccagttgtcc gctgctccgg
     4561 accctggatg tccagtgggt ggagggacta aaggatgccc agatgcggga tctcctgtcc
     4621 ccgcccacag acaacaggcc aggtcagatg gacaatcgga gcaagctccg gaacatcgtg
     4681 gagctgcgcc tggcaggcct ggacatcaca gatgcctccc tgcggctcat catccgccac
     4741 atgcccctgc tctccaagct ccacctcagt tactgtaacc acgtcaccga ccagtctatc
     4801 aacctgctca ctgctgttgg caccaccacc cgagactcct taaccgagat caacctgtct
     4861 gactgcaata aggtcactga tcagtgcctg tccttcttca aacgctgtgg aaacatctgt
     4921 catattgacc tgaggtactg caagcaagtc accaaggaag gctgtgagca gttcatagcc
     4981 gagatgtctg tgagtgtcca gtttgggcaa gtagaagaaa aactcctgca aaaactgagt
     5041 tagctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     5101 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     5161 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     5221 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     5281 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     5341 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     5401 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     5461 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     5521 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     5581 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     5641 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     5701 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     5761 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     5821 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     5881 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5941 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     6001 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     6061 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     6121 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     6181 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     6241 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     6301 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     6361 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     6421 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     6481 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     6541 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     6601 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     6661 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     6721 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     6781 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     6841 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     6901 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     6961 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     7021 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     7081 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     7141 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     7201 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     7261 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     7321 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     7381 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     7441 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     7501 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     7561 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     7621 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     7681 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     7741 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     7801 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     7861 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     7921 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     7981 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     8041 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     8101 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     8161 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     8221 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt
     8281 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     8341 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     8401 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     8461 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     8521 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     8581 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     8641 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     8701 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     8761 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     8821 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     8881 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     8941 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     9001 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     9061 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     9121 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     9181 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     9241 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     9301 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     9361 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     9421 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     9481 tgacgtc
//
