LOCUS       V019864                 8855 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019864
VERSION     V019864
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..3535
                     /label="Brd8(NM_030147)"
                     /note="Brd8(NM_030147)"
                     /gene="Brd8"
     CDS             3542..3607
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3631..4219
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4285..4333
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4535..4963
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4977..5306
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5373..6167
                     /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    6343..6464
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6513..6529)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6537..6553
                     /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(6561..6591)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6606..6627
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6915..7503)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7674..8534)
                     /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(8535..8639)
                     /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 ccatggcgac cgggacaggc aagcacaagc tgctgagtac
      721 tggcccaaca gagccatggt ccatccgaga gaagttgtgt ttagcatctt ctgtcatgag
      781 gagtggggat cagaactggg tatcagttag cagagcaatc aagccctttg cagaacctgg
      841 ccggcctcca gactggttct ctcaaaaaca ttgtgcttct cagtattctg agctcttaga
      901 gactactgag actccaaaac ggaaacgggg tgaaaaagga gaagtggtag aaactgttga
      961 agatgttatt gttcgcaaac tgactgctga gagagttgaa gaactgaaga aagtcataaa
     1021 ggagacacag gagagataca ggcgtctgaa aagagatgca gaactaatcc aagccgggca
     1081 catggacagc agactggatg agctctgcaa tgacattgca atgaaaaaga aattggaaga
     1141 ggaagaggct gaagtaaaaa ggaaggccac cgatgctgcg taccaggcgc gacaagcagt
     1201 gaaaacgcct cctcgaaggt taccgactgt gatggtccgc tctcctgtag actctgcctc
     1261 cccaggaggt gattacccac ttggagactt gactccaacc actatggaag aggctacctc
     1321 tggggtaacc cctgggactt tgccgagtac cccagtcacc tcgtttcctg ggattcctga
     1381 cacccttcct ccaggctctg cacccttaga agcccccatg accccaataa cagatgattc
     1441 accccagaaa aagatgcttg gacagaaagc aactccaccc ccctcccctc tgctgtcaga
     1501 gctcttgaag aagggcagcc tcctgcctac tagccccaga ctggtgaatg agagtgaaat
     1561 gcctgtaccc cctggccatc tgaacagcac aggggttctc ttggaggtag gaggggttct
     1621 tcccatgata catggtgggg agatacagcc aacaaccagt gctgtggcgg cctccccggc
     1681 tgcctcaggt gctcccactc tttcccggct tttagaagct gggcccacac agttcaccac
     1741 tcctcttcct tccttcacta ctgttgccag tgagccgcca gttaagcttg tgccaccccc
     1801 tgtagagtct gtgtcccagg ctaccattgt catgatgccc gcgctgccag caccatcctc
     1861 tgctgcggct gtctccactt ctgaaagtgg agctccagtg agccagcctg agccctgtgt
     1921 acccctggaa gctgtggggg atccacatac tgtgactgtt tccatggata gcaatgaaat
     1981 ctccatgatc attaattcca tcaaagaaga gtgtttccgc tcaggggtag cagaggctcc
     2041 tggtggatca aaggctccaa gcatagatgg gaaggaagac ttggatctag cggagaagat
     2101 ggatattgct gtgtcttaca caggtgaaga gttggacttt gaaacagttg gagacatcat
     2161 tgccatcatt gaggacaagg tggatgatca ccctgaagtg ctggatgtgg cagcggtaga
     2221 agcggcgtta tcgttctgtg aagagaatga tgaccctcag tccctgcctg gcccctggga
     2281 gcaccctatc cagcaggagc gcgacaagcc agtacctctg ccagcaccag agatgacagt
     2341 caaacaagag aggctagact tcgaggaatc agaaaacaaa gggctccatg acctggtgga
     2401 catcagggat tccggtgttg agattaaggt ggaacccaca gagccagagc caggcatgtc
     2461 tggggctgag atagtagctg gagttggtcc agttccaagt atggagccac cagaactcag
     2521 gagtcaagac tcagatgaag aacctagaag ttctgcagct ggagacattg gtgaggcaga
     2581 tggttccagt gggaaaggcg atgagaggcc actttcagct gtgaagacag aggcatcccc
     2641 tgagagcatg ttgtctccat cacatggctc aaatcttatt gaagatcctt tagaggcaga
     2701 gactcaacac aagtttgaaa tgtcagactc attgaaagaa gaatcaggga ctatttttgg
     2761 aagccagata aaggatgccc caggtgacga tgaggaagaa gatggagtca gtgaagcagc
     2821 tagcctagag gagcctaagg aagaggatca aggagaaggc tatttgtctg agatggataa
     2881 tgagccccct gtgagtgaga gtgatgatgg ctttagtata cataacgcca cactgcagtc
     2941 acacactctg gcagactcca tcccaagcag ccctgcctcc tcccagtttt ccgtgtgcag
     3001 tgaagatcaa gaagcaattc aggctcagaa aatatggaag aaagccatca tgcttgtatg
     3061 gagggctgca gcaaatcata ggtatgccaa tgtgttcctg caacctgtta cagatgacat
     3121 agctcctggt taccatagca ttgtacagag gcctatggat ttgtcaacta taaagaaaaa
     3181 cattgaaaat ggactgatcc gaagcacagc tgagtttcag cgtgacatca tgctgatgtt
     3241 tcagaatgct gttatgtaca atagctcaga ccatgatgtc tatcacatgg cagtagagat
     3301 gcagagagat gtcttggaac agatccagca attcttggcc acacagttga ttatgcaaac
     3361 atctgagtct ggaatcagtg ctaaaagtct ccgggggaga gactctaccc gaaaacaaga
     3421 tgcttcagag aaggacagtg tccccatggg ctctcctgcc ttccttctct ctctctttga
     3481 tgggggaacc aggggacgcc gctgtgccat tgaagcagat atgaagatga agaagctcga
     3541 ggactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga
     3601 tgacaagtag tgagggcccg atatctcgac aatcaacctc tggattacaa aatttgtgaa
     3661 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     3721 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     3781 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     3841 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     3901 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     3961 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     4021 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     4081 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     4141 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     4201 ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg aaggagacaa
     4261 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt
     4321 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     4381 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     4441 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca
     4501 gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     4561 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4621 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4681 ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4741 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4801 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4861 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat
     4921 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     4981 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5041 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     5101 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     5161 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5221 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     5281 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     5341 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     5401 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     5461 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     5521 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     5581 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5641 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5701 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5761 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5821 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5881 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5941 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     6001 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     6061 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     6121 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     6181 gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     6241 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     6301 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     6361 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     6421 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     6481 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     6541 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     6601 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6661 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     6721 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     6781 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     6841 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6901 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6961 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     7021 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     7081 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     7141 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     7201 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     7261 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     7321 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     7381 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7441 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7501 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7561 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7621 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7681 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7741 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7801 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7861 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7921 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7981 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     8041 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     8101 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     8161 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     8221 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     8281 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     8341 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     8401 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8461 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8521 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8581 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8641 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
     8701 ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     8761 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
     8821 gcttgaccga caattgcatg aagaatctgc ttagg
//
