LOCUS       V045477                 6103 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045477
VERSION     V045477
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             913..1902
                     /label="MRGPRX2(NM_001303615)"
                     /note="MRGPRX2(NM_001303615)"
                     /gene="MRGPRX2"
     CDS             1915..1932
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    1967..2191
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2237..2665
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2679..3008
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3020..3619
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    3779..3900
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(3949..3965)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    3973..3989
                     /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(3997..4027)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4042..4063
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4351..4936)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5107..5967)
                     /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(5968..6072)
                     /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 gaattcgcca ccatggatcc aaccaccccg gcctggggaa cagaaagtac aacagtgaat
      961 ggaaatgacc aagcccttct tctgctttgt ggcaaggaga ccctgatccc ggtcttcctg
     1021 atccttttca ttgccctggt cgggctggta ggaaacgggt ttgtgctctg gctcctgggc
     1081 ttccgcatgc gcaggaacgc cttctctgtc tacgtcctca gcctggccgg ggccgacttc
     1141 ctcttcctct gcttccagat tataaattgc ctggtgtacc tcagtaactt cttctgttcc
     1201 atctccatca atttccctag cttcttcacc actgtgatga cctgtgccta ccttgcaggc
     1261 ctgagcatgc tgagcaccgt cagcaccgag cgctgcctgt ccgtcctgtg gcccatctgg
     1321 tatcgctgcc gccgccccag acacctgtca gcggtcgtgt gtgtcctgct ctgggccctg
     1381 tccctactgc tgagcatctt ggaagggaag ttctgtggct tcttatttag tgatggtgac
     1441 tctggttggt gtcagacatt tgatttcatc actgcagcgt ggctgatttt tttattcatg
     1501 gttctctgtg ggtccagtct ggccctgctg gtcaggatcc tctgtggctc caggggtctg
     1561 ccactgacca ggctgtacct gaccatcctg ctcacagtgc tggtgttcct cctctgcggc
     1621 ctgccctttg gcattcagtg gttcctaata ttatggatct ggaaggattc tgatgtctta
     1681 ttttgtcata ttcatccagt ttcagttgtc ctgtcatctc ttaacagcag tgccaacccc
     1741 atcatttact tcttcgtggg ctcttttagg aagcagtggc ggctgcagca gccgatcctc
     1801 aagctggctc tccagagggc tctgcaggac attgctgagg tggatcacag tgaaggatgc
     1861 ttccgtcagg gcaccccgga gatgtcgaga agcagtctgg tgggatccct cgagcaccat
     1921 catcaccatc attgaccggt aagtttaaac cgctgatcag cctcgactgt gccttctagt
     1981 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     2041 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     2101 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     2161 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     2221 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2281 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     2341 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     2401 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     2461 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     2521 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     2581 tattcttttg atttataagg gattttggcc atttcggcct attggttaaa aaatgagctg
     2641 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     2701 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2761 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2821 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     2881 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     2941 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3001 tttgcaaaaa gctcccggga tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     3061 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     3121 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     3181 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     3241 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     3301 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     3361 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
     3421 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     3481 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     3541 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     3601 ccgcaagccc ggtgcctgat tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     3661 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     3721 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     3781 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3841 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3901 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     3961 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4021 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4081 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4141 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4201 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4261 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4321 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4381 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4441 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4501 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4561 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4621 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4681 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4741 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4801 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4861 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag
     4921 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4981 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5041 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5101 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5161 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5221 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5281 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5341 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5401 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5461 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5521 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5581 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5641 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5701 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5761 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5821 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5881 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5941 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6001 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6061 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
