LOCUS       V036862                 7128 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036862
VERSION     V036862
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             756..1808
                     /label="MSL1(NM_001012241)"
                     /note="MSL1(NM_001012241)"
                     /gene="MSL1"
     CDS             1815..1880
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1904..2492
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2558..2606
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2808..3236
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3250..3579
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3646..4440
                     /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    4616..4737
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4786..4802)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4810..4826
                     /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(4834..4864)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4879..4900
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5188..5776)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5947..6807)
                     /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(6808..6912)
                     /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 tggtaccgag ctcgaattcg ccaccatgga aaggcggatg cagctggtaa
      781 agaaggataa cgagaaagaa aggcacaagc tgtttcaggg ctatgaaact gaagagagag
      841 aggaaacaga gctatctgag aaaattaaac tggagtgcca gccggagctt tccgagacat
      901 cccagactct gcctcccaag cccttctcat gtgggcggag tggaaaggga cataaaagga
      961 aatccccatt tggaagtaca gaaagaaaga ctcctgttaa aaagctggct cctgaatttt
     1021 caaaagtcaa aacaaaaact cctaagcact ctcctattaa agaggaaccc tgtggttcct
     1081 tatctgaaac tgtttgtaaa cgtgaattga ggagccaaga aaccccagaa aagccccggt
     1141 cttcagtgga caccccacca agactctcca ctccccaaaa gggacccagc acccatccca
     1201 aggagaaagc cttctcaagt gagatagaag atttgccgta cctttccacc acagaaatgt
     1261 atttgtgtcg ttggcaccag cctcccccat caccgttacc attacgggaa tcctctccaa
     1321 agaaggagga gactgtagca aggtgtctga tgccatcaag tgttgcagga gaaacttcag
     1381 tcttggctgt tccttcttgg agggaccact cagtagagcc tctaagggac ccaaatcctt
     1441 cagacctttt ggagaacctg gatgacagtg tgttttcgaa gcggcatgca aaactggagc
     1501 tggatgagaa gagaaggaaa agatgggata ttcagaggat cagggaacaa agaattttac
     1561 agcgactgca gctcagaatg tataaaaaga aaggaattca ggaatctgag cctgaggtta
     1621 cctcattttt ccctgagcca gatgatgttg aaagtttgat gattaccccc ttcttgcctg
     1681 ttgtagcatt tggacgacca ttaccaaaat taactccaca gaattttgag ctaccctggt
     1741 tggatgagcg tagccgatgc agattggaga tccagaagaa gcaaacacct caccggacgt
     1801 gtaggaaagg atccgactac aaagaccatg acggtgatta taaagatcat gacatcgact
     1861 acaaggatga cgatgacaag tagtgagggc ccgatatctc gacaatcaac ctctggatta
     1921 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     1981 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     2041 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     2101 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     2161 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     2221 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     2281 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     2341 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     2401 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     2461 gagtcggatc tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca
     2521 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     2581 acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     2641 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     2701 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     2761 ccctgccata gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag
     2821 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2881 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2941 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca
     3001 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3061 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3121 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg
     3181 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3241 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3301 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3361 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3421 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3481 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     3541 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     3601 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3661 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3721 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3781 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     3841 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3901 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3961 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4021 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4081 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4141 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4201 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4261 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4321 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4381 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4441 gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     4501 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4561 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     4621 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4681 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     4741 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     4801 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4861 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4921 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4981 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5041 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5101 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5161 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5221 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5281 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5341 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     5401 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5461 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5521 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5581 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     5641 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5701 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5761 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5821 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5881 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5941 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6001 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6061 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6121 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6181 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6241 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6301 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6361 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6421 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6481 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6541 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6601 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6661 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6721 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6781 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6841 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6901 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     6961 cccgatcccc tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag
     7021 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     7081 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
