LOCUS       V018473                 9847 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018473
VERSION     V018473
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             928..954
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             958..984
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             988..1014
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             1042..5400
                     /label="KDM6A(NM_001291415)"
                     /note="KDM6A(NM_001291415)"
                     /gene="KDM6A"
     polyA_signal    5447..5671
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5717..6145
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6159..6488
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6555..7349
                     /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    7523..7644
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7693..7709)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7717..7733
                     /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(7741..7771)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7786..7807
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8095..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 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 ggcggcgtac ccatacgacg tacctgatta tgcaacttat
      961 ccttatgatg tacctgatta tgctagctat ccttatgatg tcccagacta cgcatcagcg
     1021 gaaaagcttg gtaccgccac catgaaatcc tgcggagtgt cgctcgctac cgccgccgct
     1081 gccgccgccg ctttcggtga tgaggaaaag aaaatggcgg cgggaaaagc gagcggcgag
     1141 agcgaggagg cgtcccccag cctgacagcc gaggagaggg aggcgctcgg cggactggac
     1201 agccgcctct ttgggttcgt gagatttcat gaagatggcg ccaggacgaa ggccctactg
     1261 ggcaaggctg ttcgctgcta tgaatctcta atcttaaaag ctgaaggaaa agtggagtct
     1321 gatttctttt gtcaattagg tcacttcaac ctcttattgg aagattatcc aaaagcatta
     1381 tctgcatacc agaggtacta cagtttacag tctgactact ggaagaatgc tgccttttta
     1441 tatggtcttg gtttggtcta cttccattat aatgcatttc agtgggcaat taaagcattt
     1501 caggaggtgc tttatgttga tcccagcttt tgtcgagcca aggaaattca tttacgactt
     1561 gggcttatgt tcaaagtgaa cacagactat gagtctagtt taaagcattt tcagttagct
     1621 ttggttgact gtaatccctg cactttgtcc aatgctgaaa ttcaatttca cattgcccac
     1681 ttatatgaaa cccagaggaa atatcattct gcaaaagaag cttatgaaca acttttgcag
     1741 acagagaatc tttctgcaca agtaaaagca actgtcttac aacagttagg ttggatgcat
     1801 cacactgtag atctcctggg agataaagcc accaaggaaa gctatgctat tcagtatctc
     1861 caaaagtcct tggaagcaga tcctaattct ggccagtcct ggtatttcct cggaaggtgc
     1921 tattcaagta ttgggaaagt tcaggatgcc tttatatctt acaggcagtc tattgataaa
     1981 tcagaagcaa gtgcagatac atggtgttca ataggtgtgc tatatcagca gcaaaatcag
     2041 cccatggatg ctttacaggc ctatatttgt gctgtacaat tggaccatgg ccatgctgca
     2101 gcctggatgg acctaggcac tctctatgaa tcctgcaacc agcctcagga tgccattaaa
     2161 tgctacttaa atgcaactag aagcaaaagt tgtagtaata cctctgcact tgcagcacga
     2221 attaagtatt tacaggctca gttgtgtaac cttccacaag gtagtctaca gaataaaact
     2281 aaattacttc ctagtattga ggaggcgtgg agcctaccaa ttcccgcaga gcttacctcc
     2341 aggcagggtg ccatgaacac agcacagcag gcatgtaaac ctcatcatcc aaatactgaa
     2401 cctgtattag gcctcagtca aacaccaatt tcacagcaat ccttgccact acacatgatt
     2461 ccttctagcc aagtagatga cctgtccagt cctgccaaga ggaaaagaac atctagtcca
     2521 acaaagaata cttctgacaa ttggagtggt ggacatgctg tgtcacatcc tccagtacag
     2581 caacaagctc attcatggtg tttgacacca cagaaattac agcatttgga acagctccgc
     2641 gcaaatagaa ataatttaaa tccagcacag aaactgatgc tggaacagct ggaaagtcag
     2701 tttgtcttaa tgcaacaaca ccaaatgaga ccaacaggag ttgcacaggt acgatctact
     2761 ggaattccta atgggccaac agctgactca tcactgccta caaactcagt ctctggccag
     2821 cagccacagc ttgctctgac cagagtgcct agcgtctctc agcctggagt ccgtcctgcc
     2881 tgccctgggc agcctttggc caatggaccc ttttctgcag gccatgttcc ctgtagcaca
     2941 tcaagaacgc tgggaagtac agacactatt ttgataggca ataatcatat aacaggaagt
     3001 ggaagtaatg gaaacgtgcc ttacctgcag cgaaacgcac tcactctacc tcataaccgc
     3061 acaaacctga ccagcagcgc agaggagccg tggaaaaacc aactatctaa ctccactcag
     3121 gggcttcaca aaggtcagag ttcacattcg gcaggtccta atggtgaacg acctctctct
     3181 tccactgggc cttcccagca tctccaggca gctggctctg gtattcagaa tcagaacgga
     3241 catcccaccc tgcctagcaa ttcagtaaca cagggggctg ctctcaatca cctctcctct
     3301 cacactgcta cctcaggtgg acaacaaggc attaccttaa ccaaagagag caagccttca
     3361 ggaaacatat tgacggtgcc tgaaacaagc aggcacactg gagagacacc taacagcact
     3421 gccagtgtcg agggacttcc taatcatgtc catcagatga cggcagatgc tgtttgcagt
     3481 cctagccatg gagattctaa gtcaccaggt ttactaagtt cagacaatcc tcagctctct
     3541 gccttgttga tgggaaaagc caataacaat gtgggtactg gaacctgtga caaagtcaat
     3601 aacatccacc cagctgttca tacaaagact gataactctg ttgcctcttc accatcttca
     3661 gccatttcaa cagcaacacc ttctccaaaa tccactgagc agacaaccac aaacagtgtt
     3721 accagcctta acagccctca cagtgggcta cacacaatta atggagaagg gatggaagaa
     3781 tctcagagcc ccatgaaaac agatctgctt ctggttaacc acaaacctag tccacagatc
     3841 ataccatcaa tgtctgtgtc catatacccc agctcagcag aagttctgaa ggcatgcagg
     3901 aatctaggta aaaatggctt atctaacagt agcattttgt tggataaatg tccacctcca
     3961 agaccaccat cttcaccata ccctcccttg ccaaaggaca agttgaatcc acctacacct
     4021 agtatttact tggaaaataa acgtgatgct ttctttcctc cattacatca attttgtaca
     4081 aatccgaaca accctgttac agtaatacgt ggccttgctg gagctcttaa gttagacctg
     4141 ggacttttct ctactaaaac tttggtggaa gctaacaatg aacatatggt agaagtgagg
     4201 acacagttgt tgcagccagc agatgaaaac tgggatccca ctggaacaaa gaaaatctgg
     4261 cattgtgaaa gtaatagatc tcatactaca attgctaaat atgcacagta ccaggcctcc
     4321 tcattccagg aatcattgag agaagaaaat gaaaaaagaa gtcatcataa agaccactca
     4381 gatagtgaat ctacatcgtc agataattct gggaggagga ggaaaggacc ctttaaaacc
     4441 ataaagtttg ggaccaatat tgacctatct gatgacaaaa agtggaagtt gcagctacat
     4501 gagctgacta aacttcctgc ttttgtgcgt gtcgtatcag caggaaatct tctaagccat
     4561 gttggtcata ccatattggg catgaacaca gttcaactat acatgaaagt tccagggagc
     4621 agaacaccag gtcatcagga aaataacaac ttctgttcag ttaacataaa tattggccca
     4681 ggtgactgtg aatggtttgt tgttcctgaa ggttactggg gtgttctgaa tgacttctgt
     4741 gaaaaaaata atttgaattt cctaatgggt tcttggtggc ccaatcttga agatctttat
     4801 gaagcaaatg ttccagtgta taggtttatt cagcgacctg gagatttggt ctggataaat
     4861 gcaggcactg ttcattgggt tcaggctatt ggctggtgca acaacattgc ttggaatgtt
     4921 ggtccactta cagcctgcca gtataaattg gcagtggaac ggtacgaatg gaacaaattg
     4981 caaagtgtga agtcaatagt acccatggtt catctttcct ggaatatggc acgaaatatc
     5041 aaggtctcag atccaaagct ttttgaaatg attaagtatt gtcttctaag aactctgaag
     5101 caatgtcaga cattgaggga agctctcatt gctgcaggaa aagagattat atggcatggg
     5161 cggacaaaag aagaaccagc tcattactgt agcatttgtg aagtggaggt ttttgatctg
     5221 ctttttgtca ctaatgagag taattcacga aagacctaca tagtacattg ccaagattgt
     5281 gcacgaaaaa caagcggaaa cttggaaaac tttgtggtgc tagaacagta caaaatggag
     5341 gacctgatgc aagtctatga ccaatttaca ttagctcctc cattaccatc cgcctcatct
     5401 tgactcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     5461 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     5521 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     5581 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     5641 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     5701 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     5761 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     5821 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     5881 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     5941 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     6001 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     6061 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     6121 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     6181 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     6241 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     6301 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     6361 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     6421 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     6481 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     6541 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     6601 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     6661 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     6721 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     6781 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     6841 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     6901 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     6961 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     7021 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     7081 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     7141 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     7201 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     7261 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     7321 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     7381 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     7441 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     7501 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     7561 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     7621 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     7681 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     7741 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     7801 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     7861 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     7921 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     7981 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     8041 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     8101 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     8161 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     8221 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     8281 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     8341 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     8401 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     8461 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     8521 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     8581 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 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 tgacgtc
//
