LOCUS       V030773                 6856 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030773
VERSION     V030773
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"
     CDS             925..2808
                     /label="ASH2L(NM_004674)"
                     /note="ASH2L(NM_004674)"
                     /gene="ASH2L"
     CDS             2821..2886
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2921..3145
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3191..3619
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3633..3962
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3974..4372
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /note="confers resistance to blasticidin"
                     /product="blasticidin S deaminase"
                     /transl_table=1
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG*"
                     /label="BSD"
     polyA_signal    4532..4653
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4702..4718)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4726..4742
                     /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(4750..4780)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4795..4816
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5104..5689)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5860..6720)
                     /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(6721..6825)
                     /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 aagctttcta gagaattcgc caccatggcg gcggcaggag caggacctgg ccaggaagcg
      961 ggtgccgggc ctggcccagg agcggtcgca aatgcaacag gggcagaaga gggggagatg
     1021 aagccggtgg cagcgggagc agccgctcct cctggagagg ggatctctgc tgctccgaca
     1081 gttgagccca gttccgggga ggctgaaggc ggggaggcaa acttggtcga tgtaagcggt
     1141 ggcttggaga cagaatcatc taatggaaaa gatacactag aaggtgctgg ggatacatca
     1201 gaggtgatgg atactcaggc gggctccgtg gatgaagaga atggccgaca gttgggtgag
     1261 gtagagctgc aatgtgggat ttgtacaaaa tggttcacgg ctgacacatt tggcatagat
     1321 acctcatcct gtctaccttt catgaccaac tacagttttc attgcaacgt ctgccatcac
     1381 agtgggaata cctatttcct ccggaagcaa gcaaacttga aggaaatgtg ccttagtgct
     1441 ttggccaacc tgacatggca gtcccgaaca caggatgaac atccgaagac aatgttctcc
     1501 aaagataagg atattatacc atttattgat aaatactggg agtgcatgac aaccagacag
     1561 agacctggga aaatgacttg gccaaataac attgttaaaa caatgagtaa agaaagagat
     1621 gtattcttgg taaaggaaca cccagatcca ggcagtaaag atccagaaga agattacccc
     1681 aaatttggac ttttggatca ggaccttagt aacattggtc ctgcttatga caaccaaaaa
     1741 cagagcagtg ctgtgtctac tagtgggaat ttaaatgggg gaattgcagc aggaagcagc
     1801 ggaaaaggac gaggagccaa gcgcaaacag caggatggag ggaccacagg gaccaccaag
     1861 aaggcccgga gtgacccttt gttttctgct cagcgccttc cccctcatgg ctacccattg
     1921 gaacacccgt ttaacaaaga tggctatcgg tatattctag ctgagcctga tccgcacgcc
     1981 cctgaccccg agaagctgga acttgactgc tgggcaggaa aacctattcc tggagacctc
     2041 tacagagcct gcttgtatga acgggttttg ttagccctac atgatcgagc tccccagtta
     2101 aagatctcag atgaccggct gactgtggtt ggagagaagg gctactctat ggtgagggcc
     2161 tctcatggag tacggaaagg tgcctggtat tttgaaatca ctgtggatga gatgccacca
     2221 gataccgctg ccagactggg ttggtcccag cccctaggaa accttcaagc tcctttaggt
     2281 tatgataaat ttagctattc ttggcggagc aaaaagggaa ccaagttcca ccagtccatt
     2341 ggcaaacact actcttctgg ctatggacag ggagacgtcc tgggatttta tattaatctt
     2401 cctgaagaca cagagacagc caagtcattg ccagacacat acaaagataa ggctttgata
     2461 aaattcaaga gttatttgta ttttgaggaa aaagactttg tggataaagc agagaagagc
     2521 ctgaagcaga ctccccatag tgagataata ttttataaaa atggtgtcaa tcaaggtgtg
     2581 gcttacaaag atatttttga gggggtttac ttcccagcca tctcactgta caagagctgc
     2641 acggtttcca ttaactttgg accatgcttc aagtatcctc cgaaggatct cacttaccgc
     2701 cctatgagtg acatgggctg gggcgccgtg gtagagcaca ccctggctga cgtcttgtat
     2761 cacgtggaga cagaagtgga tgggaggcgc agtcccccat gggaacccct cgagaccggt
     2821 gactacaaag accatgacgg tgattataaa gatcatgaca tcgattacaa ggatgacgat
     2881 gacaagtgac cggtaagttt aaaccgctga tcagcctcga ctgtgccttc tagttgccag
     2941 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     3001 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     3061 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     3121 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     3181 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3241 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     3301 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     3361 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     3421 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3481 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     3541 tttgatttat aagggatttt ggccatttcg gcctattggt taaaaaatga gctgatttaa
     3601 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3661 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3721 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3781 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3841 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3901 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3961 aaaagctccc gggatggcca agcctttgtc tcaagaagaa tccaccctca ttgaaagagc
     4021 aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct
     4081 ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg
     4141 tgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat
     4201 cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt
     4261 gcttctcgat ctgcatcctg ggatcaaagc catagtgaag gacagtgatg gacagccgac
     4321 ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggct aattcgaaat
     4381 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4441 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4501 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     4561 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4621 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     4681 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4741 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4801 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4861 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4921 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4981 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5041 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5101 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5161 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5221 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5281 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5341 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5401 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5461 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5521 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5581 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5641 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5701 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5761 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5821 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5881 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5941 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6001 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6061 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6121 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6181 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6241 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6301 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6361 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6421 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6481 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6541 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6601 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6661 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6721 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6781 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6841 agtgccacct gacgtc
//
