LOCUS       V032889                 6446 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032889
VERSION     V032889
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             938..961
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             968..1933
                     /label="MRGPRX1(NM_147199)"
                     /note="MRGPRX1(NM_147199)"
                     /gene="MRGPRX1"
     polyA_signal    2013..2237
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2283..2711
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2725..3054
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3121..3915
                     /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    4089..4210
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4259..4275)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4283..4299
                     /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(4307..4337)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4352..4373
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4661..5246)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5417..6277)
                     /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(6278..6382)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttgccacca tgaagaccat
      901 catcgccctc agctacatct tttgtctggt cttcgcggat tacaaggatg acgacgataa
      961 gggatccatg gatccaacca tctcaacctt ggacacagaa ctgacaccaa tcaacggaac
     1021 tgaggagact ctttgctaca agcagacctt gagcctcacg gtgctgacgt gcatcgtttc
     1081 ccttgtcggg ctgacaggaa acgcagttgt gctctggctc ctgggctgcc gcatgcgcag
     1141 gaacgccttc tccatctaca tcctcaactt ggccgcagca gacttcctct tcctcagcgg
     1201 ccgccttata tattccctgt taagcttcat cagtatcccc cataccatct ctaaaatcct
     1261 ctatcctgtg atgatgtttt cctactttgc aggcctgagc tttctgagtg ccgtgagcac
     1321 cgagcgctgc ctgtccgtcc tgtggcccat ctggtaccgc tgccaccgcc ccacacacct
     1381 gtcagcggtg gtgtgtgtcc tgctctgggc cctgtccctg ctgcggagca tcctggagtg
     1441 gatgttatgt ggcttcctgt tcagtggtgc tgattctgct tggtgtcaaa catcagattt
     1501 catcacagtc gcgtggctga tttttttatg tgtggttctc tgtgggtcca gcctggtcct
     1561 gctgatcagg attctctgtg gatcccggaa gataccgctg accaggctgt acgtgaccat
     1621 cctgctcaca gtactggtct tcctcctctg tggcctgccc tttggcattc agtttttcct
     1681 atttttatgg atccacgtgg acagggaagt cttattttgt catgttcatc tagtttctat
     1741 tttcctgtcc gctcttaaca gcagtgccaa ccccatcatt tacttcttcg tgggctcctt
     1801 taggcagcgt caaaataggc agaacctgaa gctggttctc cagagggctc tgcaggacgc
     1861 gtctgaggtg gatgaaggtg gagggcagct tcctgaggaa atcctggagc tgtcgggaag
     1921 cagattggag cagtgagaat tctgcagata tccagcacag tggcggccgc tcgagtctag
     1981 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     2041 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2101 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     2161 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     2221 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     2281 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2341 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2401 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2461 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     2521 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2581 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     2641 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     2701 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     2761 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     2821 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2881 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     2941 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     3001 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     3061 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     3121 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     3181 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     3241 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     3301 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     3361 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     3421 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     3481 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     3541 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     3601 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     3661 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     3721 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     3781 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     3841 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     3901 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     3961 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4021 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4081 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4141 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4201 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     4261 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4321 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4381 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4441 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4501 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4561 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4621 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4681 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4741 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4801 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4861 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4921 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4981 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5041 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5101 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5161 gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga
     5221 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5281 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5341 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5401 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5461 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5521 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5581 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5641 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5701 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5761 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5821 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5881 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5941 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6001 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6061 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6121 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6181 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6241 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6301 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6361 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     6421 cgggagatct cccgatcccc tatggt
//
