LOCUS       V021161                 7885 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021161
VERSION     V021161
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"
     CDS             683..2533
                     /label="Brd9(NM_001378947)"
                     /note="Brd9(NM_001378947)"
                     /gene="Brd9"
     CDS             2572..2637
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2661..3249
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3315..3363
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3565..3993
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4007..4336
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4403..5197
                     /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    5373..5494
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5543..5559)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5567..5583
                     /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(5591..5621)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5636..5657
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5945..6533)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6704..7564)
                     /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(7565..7669)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatgggcaa aaagcacaag aagcacaagg cggaatggcg
      721 ctcgtcctac gaagattata cagacacgcc actggagaag cctctgaagc tggtgctcaa
      781 ggtgggagga agtgaagtga cagagctctc aggatctggc cacgactcca gctactacga
      841 cgatcgctca gaccacgaac gggagagaca cagagaaaag aagaaaaaga agaagaaaaa
      901 gtcagagaag gagaagcacc tcgatgagga ggagaggagg aagcggaagg aagagaagaa
      961 acggaaacgg gagaaggaac actgcgactc agagggggag gctgatgctt tcgaccctgg
     1021 aaagaaggtg gaggtggagc cacccccaga ccgaccagtg agagcctgcc gaacacagcc
     1081 agctgagaac gagagcacac ctatccagag gcttctggaa cacttcctcc gccagctaca
     1141 gagaaaagat cctcatggat tttttgcttt tcctgttacg gatgcaattg ctcctgggta
     1201 ttcaatgata ataaaacatc ctatggactt tggcacgatg aaagacaaga ttgtagctaa
     1261 tgaatataaa tcagtcacag aatttaaggc agatttcaaa ttaatgtgtg ataatgcgat
     1321 gacgtacaat agaccagaca ccgtgtacta caaattagcc aagaagatcc tgcacgcggg
     1381 ctttaagatg atgagcaaag agcggctgtt agctttgaag cgcagcatgt cgtttatgca
     1441 ggacatggat ttctctcagc aggcagctct cttgggcagt gaagacccag cagctgagga
     1501 acctgttccc gaggttgtcc cagtgcaagt agaaactacc aagaaatcca aaaagccgag
     1561 tagagaagtt atcagctgca tgtttgagcc tgaagggaat gcctgcagcc tgacagacag
     1621 cacggcagag gagcatgtgc tagccctggt agagcacgca gctgatgagg ctcgggacag
     1681 gattaaccgg tttctcccgg gtggcaagat ggggtacctg aagaagcttg gagatggaag
     1741 tctgctctac agcgtggtga acgcacctga gcctgatgct gatgaggagg agacacaccc
     1801 tgtggacctg agttcactgt ctagcaagtt gctcccaggt tttacaacat tgggtttcaa
     1861 agatgaaaga agaaataaag tcacattcct ctccagtgcc agcactgcac tttcaatgca
     1921 gaacaactct gtgtttgggg acctgaagtc agatgagatg gagcttctgt attccgccta
     1981 tggagatgag actggtgtgc agtgtgcact gagcctgcag gaattcgtga aggatgctgg
     2041 aagctatagc aagaagatgg tagatgacct cctggaccaa atcacaggtg gtgatcactc
     2101 aaggatgatc ttccagctga agcagaggag gagcatcccc atgagacctg cagatgagat
     2161 gaaggttggg gatccactgg gagagagtgg tggccctgtt ctggacttca tgtcaatgaa
     2221 acagtatcct gatgtctccc tggatgtgtc catgctcagc tctctcggga aagtaaagaa
     2281 ggagctggac catgaagata gccacttgaa cttggatgag acagccaggc tcctgcagga
     2341 cttacacgaa gcacaagcag agcgaggagg ctctcggcca tcctccaacc ttagctctct
     2401 gtccactgcc tctgagaggg agcatcctcc tccaggaagt ccttctcgcc ttagtgttgg
     2461 ggagcagccg gatgtcgccc acgaccctta tgaattcctt cagtctccag aacctgcagc
     2521 tcctgccaag aacaattctg cagatatcca gcacagtggc ggccgctcga ggactacaaa
     2581 gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag
     2641 tgagggcccg atatctcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     2701 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     2761 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     2821 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     2881 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     2941 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     3001 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga
     3061 cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     3121 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     3181 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     3241 cctccccgcc tggaaacggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     3301 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg
     3361 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     3421 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     3481 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca gatctgcgca
     3541 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3601 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3661 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3721 gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3781 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3841 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3901 tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa
     3961 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     4021 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4081 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4141 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4201 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4261 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4321 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     4381 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4441 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4501 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4561 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     4621 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4681 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4741 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4801 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4861 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4921 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4981 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     5041 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     5101 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5161 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa
     5221 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5281 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5341 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5401 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5461 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5521 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5581 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5641 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5701 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5761 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5821 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5881 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5941 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6001 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6061 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6121 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     6181 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6241 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6301 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6361 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     6421 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6481 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6541 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6601 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6661 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6721 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6781 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6841 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6901 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6961 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7021 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7081 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7141 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7201 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7261 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7321 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7381 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7441 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7501 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7561 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7621 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7681 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtcgactct
     7741 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     7801 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     7861 caattgcatg aagaatctgc ttagg
//
