LOCUS       V042635                10032 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042635
VERSION     V042635
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             744..4712
                     /label="Kdm3a(NM_173001)"
                     /note="Kdm3a(NM_173001)"
                     /gene="Kdm3a"
     CDS             4719..4784
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4808..5396
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5462..5510
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5712..6140
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6154..6483
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6550..7344
                     /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    7520..7641
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7690..7706)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7714..7730
                     /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(7738..7768)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7783..7804
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8092..8680)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8851..9711)
                     /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(9712..9816)
                     /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 tggtaccgcc accatggtgc tcacgctcgg agaaagttgg ccagtattgg
      781 tggggaagcg attcctcagt ctgtccgcag ccgaaggcaa cgaaggcggc caggacaact
      841 gggacttgga gcgcgttgcc gagtggccct ggctgtcggg gaccattcga gctgtttccc
      901 acaccgacgt tactaagaaa gacttgaagg tgtgtgtgga gtttgatggg gagtcttgga
      961 gaaagagaag atggatagat gtctacagcc ttcagagaaa agcattttta gtagagcata
     1021 acctggtttt ggcagaacga aaatcacctg aagttcctga gcaagttatt cagtggcctg
     1081 caataatgta caaatctctt ctagacaaag ctggcttggg agccataact tctgttcggt
     1141 ttcttggaga tcaacaaagt gtatttgttt ccaaagacct tttgaaacct atacaggatg
     1201 ttaacagtct tcggctttcc cttactgata atcagacagt cagtaaggaa tttcaagctt
     1261 tgattgtaaa acatttggat gaaagccatc ttttacaagg tgacaagaac cttgttggtt
     1321 cagaagtaaa aatttatagc ttggacccat ctactcagtg gttttcagca actgttgtac
     1381 atggaaaccc atcatccaaa actcttcaag tcaactgtga ggagattcca gcactgaaaa
     1441 ttgtcgaccc agcactgatt catgttgaag ttgtacatga caactttgtg acatgtggta
     1501 attctacaag aactggagct gtaaaacgca agtcttctga gaataacgga agttcggttt
     1561 ctaaacaagc aaaatcttgt tctgaggcct ctcccagtat gtgtcctgta cagtctgttc
     1621 ccacaacagt gtttaaggag atcctgcttg gctgtactgc agcaactcca tctagcaagg
     1681 acccaagaca gcaaaatact ccccaggcag ccaattctcc acctaacatt ggagcaaaac
     1741 ttcctcaagg atgtcataag cagaacttac cagaagaact ttcttcctgt ctaaacacaa
     1801 aacctgaagt accgagaaca aaaccagatg tctgcaaaga aggattactt tcttcaaaat
     1861 cttctcaggt tggagctgga gacttgaaaa ttctgagtga gcccaaaggt agctgtatcc
     1921 agcctaaaac aaacactgat caggagagca gactggagtc tgctccacag ccagtcactg
     1981 gccttccaaa ggagtgcttg cctgcaaaga cttcctctaa ggcagaactg gacattgcca
     2041 ccactcctga actgcagaag catctagaac atgcagcttc cacatccgat gacctttcag
     2101 ataagccaga agtgaaagca ggtgtcacta gccttaatag ttgtgcagaa aagaaggtcg
     2161 aaccttcaca tttaggttcc cagtcacaga atttaaagga aacttcagta aaagtagata
     2221 atgaaagctg ttgtacaaga agcagtaata aaacccagac tcccccagcc cggaagtcag
     2281 ttttgacaga cccagataaa gtcaggaagc tgcagcagag cggagaggcc tttgttcagg
     2341 atgactcctg tgttaacatc gtggcacagc tgcccaagtg tcgggagtgt cgactagaca
     2401 gcctgcgcaa ggataaggac cagcagaagg actctcctgt gttttgtcgc tttttccact
     2461 tcaggagatt acaattcaac aagcatggtg tgttgcgggt agaaggcttc ttaacaccaa
     2521 acaagtatga cagtgaagcg attggcttgt ggctgccttt gaccaaaaat gttgtgggga
     2581 ctgatttgga cacagcaaaa tatatcctgg ccaatattgg agaccacttc tgtcaaatgg
     2641 tgatttctga gaaggaagct atgtcgacta ttgagccaca cagacaggtt gcttggaaac
     2701 gagctgtcaa aggagttaga gaaatgtgtg atgtgtgtga cacaaccatt ttcaacctgc
     2761 actgggtgtg ccctcggtgt gggtttggag tatgtgtaga ttgctaccgg atgaagagga
     2821 agaactgcca acagggtgct gcctacaaga ctttctcttg gataaggtgt gtgaagagtc
     2881 agatacatga gcctgagaac ctgatgccca cacagattat tcctggcaaa gccctctacg
     2941 atgttggaga cattgtgcat tctgtcagag caaaatgggg cataaaggcc aattgtccct
     3001 gctccaacag gcagttcaag ctcttctcaa agccagcctt aaaggaagac ctgaaacaga
     3061 catccttgtc tggagaaaaa ccaactcttg ggaccatggt ccagcaaagt tcccctgttt
     3121 tggagccagt ggctgtgtgc ggggaagcag cctccaagcc agccagcagc gtgaagccca
     3181 cctgtcccac cagcacttca cctttaaact ggctagctga ccttaccagt gggaatgtca
     3241 acaaggagaa taaggaaaaa cagctgacta tgccaatttt aaagaatgaa atcaaatgcc
     3301 ttccaccctt gccccctctg aacaagccca gcacagtcct ccatactttt aacagcacca
     3361 tcttgacacc tgtgagcaac aataattcag gtttccttag aaatcttttg aattcatcca
     3421 cagcaaagac agaaaatgga ttgaaaaaca cacccaaaat tcttgatgac atctttgcct
     3481 ctttggtgca aaacaagact tcttctgatt catccaagag gcctcaagga ctgacaatca
     3541 agcctagcat tcttggcttt gacactcctc actactggct gtgtgacaac cgcctgctgt
     3601 gcttgcaaga ccccaacaat aagagcaatt ggaatgtttt tagggaatgc tggaaacaag
     3661 ggcagccagt gatggtgtcg ggcgtgcatc ataaattaaa cactgaactc tggaaacccg
     3721 agtccttcag aaaagagttt ggtgagcagg aagtagacct agtcaattgt aggaccaatg
     3781 aaatcatcac tggagccaca gtcggagact tctgggatgg atttgaagat gttccaaacc
     3841 gtttgaaaaa cgacaaagaa aaagaaccaa tggtgttgaa acttaaggac tggccgccag
     3901 gagaagactt cagagacatg atgccttcca ggtttgatga tctgatggcc aacattccac
     3961 tgcctgagta caccaggcga gatggcaaac tgaacctggc ttccagactg ccaaactact
     4021 ttgtacggcc agacctgggc cccaagatgt acaatgctta tggattgatc actccagagg
     4081 atcggaaata tgggaccaca aatcttcact tagatgtatc tgatgcagcc aatgtcatgg
     4141 tttatgtggg aattcccaaa ggacagtgtg aacaagaaga agaagtcctt agaaccatcc
     4201 aagatggaga ttctgatgaa ctcacaatca agagatttat tgaaggaaaa gagaagccag
     4261 gagccctttg gcacatatat gctgctaaag acacagagaa gataagagaa ttccttaaaa
     4321 aggtatcaga ggagcagggt caagacaacc ctgcagacca tgaccctatc cacgatcaga
     4381 gctggtattt agaccgatcg ctgagaaagc gcctctatca agagtacggc gtgcaaggct
     4441 gggctattgt acagtttctt ggggatgtgg tgtttatccc agcaggagcg ccacatcagg
     4501 ttcataactt atacagctgt atcaaagtgg ctgaagactt tgtgtctcca gagcatgtta
     4561 aacactgctt ctggcttact caggaattcc gttacttgtc acagactcat accaaccatg
     4621 aagataaatt gcaggtgaaa aatgttatct accatgcagt gaaagatgca gttgctatgc
     4681 tgaaagccag tgaatccagt ttgggcaaac ctggatccga ctacaaagac catgacggtg
     4741 attataaaga tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgata
     4801 tctcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     4861 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     4921 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     4981 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     5041 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     5101 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     5161 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     5221 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     5281 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     5341 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     5401 aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     5461 acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg
     5521 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     5581 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     5641 cgcagccaac gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag
     5701 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     5761 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     5821 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
     5881 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     5941 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     6001 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     6061 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
     6121 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     6181 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     6241 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     6301 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     6361 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     6421 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     6481 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     6541 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     6601 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     6661 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     6721 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     6781 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     6841 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     6901 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     6961 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     7021 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     7081 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     7141 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     7201 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     7261 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     7321 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc
     7381 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     7441 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     7501 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     7561 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     7621 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     7681 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     7741 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     7801 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     7861 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     7921 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     7981 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     8041 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     8101 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     8161 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     8221 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     8281 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     8341 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     8401 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     8461 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     8521 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     8581 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     8641 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     8701 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     8761 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     8821 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     8881 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     8941 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     9001 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     9061 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     9121 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     9181 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     9241 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     9301 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     9361 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     9421 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     9481 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     9541 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     9601 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     9661 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     9721 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     9781 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     9841 tgacgtcgac ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc
     9901 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     9961 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
    10021 aatctgctta gg
//
