LOCUS       V044219                 6188 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044219
VERSION     V044219
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..2011
                     /label="TAS2R7(NM_023919)"
                     /note="TAS2R7(NM_023919)"
                     /gene="TAS2R7"
     CDS             2018..2050
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /transl_table=1
                     /translation="QPELAPEDPED"
                     /label="HSV tag"
     polyA_signal    2156..2380
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     protein_bind    2664..2711
                     /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             2719..3741
                     /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    3871..3992
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4041..4057)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4065..4081
                     /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(4089..4119)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4134..4155
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4443..5031)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5202..6062)
                     /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(6063..6167)
                     /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 gcagataaag tgcagactac
     1081 tttattgttc ttagcagttg gagagttttc agtggggatc ttagggaatg cattcattgg
     1141 attggtaaac tgcatggact gggtcaagaa gaggaaaatt gcctccattg atttaatcct
     1201 cacaagtctg gccatatcca gaatttgtct attgtgcgta atactattag attgttttat
     1261 attggtgcta tatccagatg tctatgccac tggtaaagaa atgagaatca ttgacttctt
     1321 ctggacacta accaatcatt taagtatctg gtttgcaacc tgcctcagca tttactattt
     1381 cttcaagata ggtaatttct ttcacccact tttcctctgg atgaagtgga gaattgacag
     1441 ggtgatttcc tggattctac tggggtgcgt ggttctctct gtgtttatta gccttccagc
     1501 cactgagaat ttgaacgctg atttcaggtt ttgtgtgaag gcaaagagga aaacaaactt
     1561 aacttggagt tgcagagtaa ataaaactca acatgcttct accaagttat ttctcaacct
     1621 ggcaacgctg ctcccctttt gtgtgtgcct aatgtccttt ttcctcttga tcctctccct
     1681 gcggagacat atcaggcgaa tgcagctcag tgccacaggg tgcagagacc ccagcacaga
     1741 agcccatgtg agagccctga aagctgtcat ttccttcctt ctcctcttta ttgcctacta
     1801 tttgtccttt ctcattgcca cctccagcta ctttatgcca gagacggaat tagctgtgat
     1861 ttttggtgag tccatagctc taatctaccc ctcaagtcat tcatttatcc taatactggg
     1921 gaacaataaa ttaagacatg catctctaaa ggtgatttgg aaagtaatgt ctattctaaa
     1981 aggaagaaaa ttccaacaac ataaacaaat caccggtcag cccgaactgg ctcccgagga
     2041 tccagaggac taggatccac tagtccagtg tggtggaatt ctgcagatat ccagcacagt
     2101 ggcggccgct cgatcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg
     2161 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     2221 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     2281 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     2341 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     2401 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2461 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     2521 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2581 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2641 tagggtgatg gttcacgtac ctagaagttc ctattccgaa gttcctattc tctagaaagt
     2701 ataggaactt ccttggccaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg
     2761 atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt
     2821 gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat
     2881 ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg
     2941 gaagtgcttg acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca
     3001 cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc
     3061 gcggaggcca tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca
     3121 ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct
     3181 gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg
     3241 caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg
     3301 cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt
     3361 gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg
     3421 aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag
     3481 cttgcaggat cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat
     3541 cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca
     3601 atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc
     3661 gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact
     3721 cgtccgaggg caaaggaata gcacgtacta cgagatttcg attccaccgc cgccttctat
     3781 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     3841 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     3901 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3961 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     4021 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     4081 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     4141 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4201 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4261 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4321 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4381 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4441 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4501 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4561 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4621 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4681 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4741 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4801 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4861 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4921 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4981 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5041 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5101 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5161 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5221 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5281 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5341 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5401 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5461 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5521 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5581 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5641 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5701 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5761 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5821 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5881 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5941 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6001 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6061 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6121 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6181 aaagtgcc
//
