LOCUS       V044535                 6233 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044535
VERSION     V044535
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        245..624
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        625..828
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        873..891
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     regulatory      911..920
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             1058..2056
                     /label="TAS2R38(NM_176817)"
                     /note="TAS2R38(NM_176817)"
                     /gene="TAS2R38"
     CDS             2063..2095
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /transl_table=1
                     /translation="QPELAPEDPED"
                     /label="HSV tag"
     polyA_signal    2201..2425
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     protein_bind    2709..2756
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2μ plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core
                     sequence TCTAGAAA (Turan and Bode, 2011)."
                     /label="FRT"
     CDS             2764..3786
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /note="confers resistance to hygromycin"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /transl_table=1
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     *"
                     /label="HygR"
     polyA_signal    3916..4037
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4086..4102)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4110..4126
                     /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(4134..4164)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4179..4200
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4488..5076)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5247..6107)
                     /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(6108..6212)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc
       61 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
      121 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
      181 aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg
      241 cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
      301 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      361 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      421 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      481 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      541 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      601 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
      661 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      721 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      781 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact
      841 agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa
      901 gctggctagc gccaccatgg ccgctgttac ctatccttca tccgtgccta cgaccttgga
      961 ccctgggaat gcatcctcag cctggcccct ggacacgtcc ctggggaatg catctgctgg
     1021 cactagcctg gcaggactgg ctgtcagtgg cctcgagatg ttgactctaa ctcgcatccg
     1081 cactgtgtcc tatgaagtca ggagtacatt tctgttcatt tcagtcctgg agtttgcagt
     1141 ggggtttctg accaatgcct tcgttttctt ggtgaatttt tgggatgtag tgaagaggca
     1201 ggcactgagc aacagtgatt gtgtgctgct gtgtctcagc atcagccggc ttttcctgca
     1261 tggactgctg ttcctgagtg ctatccagct tacccacttc cagaagttga gtgaaccact
     1321 gaaccacagc taccaagcca tcatcatgct atggatgatt gcaaaccaag ccaacctctg
     1381 gcttgctgcc tgcctcagcc tgctttactg ctccaagctc atccgtttct ctcacacctt
     1441 cctgatctgc ttggcaagct gggtctccag gaagatctcc cagatgctcc tgggtattat
     1501 tctttgctcc tgcatctgca ctgtcctctg tgtttggtgc ttttttagca gacctcactt
     1561 cacagtcaca actgtgctat tcatgaataa caatacaagg ctcaactggc agattaaaga
     1621 tctcaattta ttttattcct ttctcttctg ctatctgtgg tctgtgcctc ctttcctatt
     1681 gtttctggtt tcttctggga tgctgactgt ctccctggga aggcacatga ggacaatgaa
     1741 ggtctatacc agaaactctc gtgaccccag cctggaggcc cacattaaag ccctcaagtc
     1801 tcttgtctcc tttttctgct tctttgtgat atcatcctgt gctgccttca tctctgtgcc
     1861 cctactgatt ctgtggcgcg acaaaatagg ggtgatggtt tgtgttggga taatggcagc
     1921 ttgtccctct gggcatgcag ccatcctgat ctcaggcaat gccaagttga ggagagctgt
     1981 gatgaccatt ctgctctggg ctcagagcag cctgaaggta agagccgacc acaaggcaga
     2041 ttcccggaca ctgtgcaccg gtcagcccga actggctccc gaggatccag aggactagga
     2101 tccactagtc cagtgtggtg gaattctgca gatatccagc acagtggcgg ccgctcgatc
     2161 gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
     2221 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     2281 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     2341 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     2401 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     2461 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2521 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2581 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2641 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2701 cgtacctaga agttcctatt ccgaagttcc tattctctag aaagtatagg aacttccttg
     2761 gccaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac
     2821 agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat
     2881 gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat
     2941 cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt
     3001 ggggaattca gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg
     3061 caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggccatggat
     3121 gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga
     3181 atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat
     3241 cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag
     3301 ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc
     3361 tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg
     3421 atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct
     3481 tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccg
     3541 cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac
     3601 ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga
     3661 gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc
     3721 tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag
     3781 gaatagcacg tactacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     3841 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     3901 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3961 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     4021 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     4081 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4141 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4201 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4261 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4321 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4381 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4441 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4501 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4561 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4621 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4681 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4741 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4801 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4861 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4921 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4981 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5041 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5101 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5161 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5221 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5281 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5341 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5401 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5461 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5521 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5581 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5641 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5701 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5761 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5821 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5881 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5941 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6001 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6061 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6121 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6181 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gcc
//
