LOCUS       V026447                 8592 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026447
VERSION     V026447
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..3272
                     /label="TAS1R2(NM_152232)"
                     /note="TAS1R2(NM_152232)"
                     /gene="TAS1R2"
     CDS             3279..3344
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3368..3956
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4022..4070
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4272..4700
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4714..5043
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5110..5904
                     /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    6080..6201
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6250..6266)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6274..6290
                     /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(6298..6328)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6343..6364
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6652..7240)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7411..8271)
                     /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(8272..8376)
                     /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 ccaccatggg gcccagggca aagaccatct
      781 cctccctgtt cttcctccta tgggtcctgg ctgagccggc tgagaactcg gacttctacc
      841 tgcctgggga ttacctcctg ggtggcctct tctccctcca tgccaacatg aagggcattg
      901 ttcaccttaa cttcctgcag gtgcccatgt gcaaggagta tgaagtgaag gtgataggct
      961 acaacctcat gcaggccatg cgctttgcgg tggaggagat caacaatgac agcagcctgc
     1021 tgcctggtgt gctgctgggc tatgagatcg tggatgtgtg ctacatctcc aacaatgtcc
     1081 agccggtgct ctacttcctg gcacacgagg acaacctcct tcccatccaa gaggactaca
     1141 gtaactacat ttcccgtgtg gtggctgtca ttggccctga caactccgag tctgtcatga
     1201 ctgtggccaa cttcctctcc ctatttctcc ttccacagat cacctacagc gccatcagcg
     1261 atgagctgcg agacaaggtg cgcttcccgg ctttgctgcg taccacaccc agcgccgacc
     1321 accacatcga ggccatggtg cagctgatgc tgcacttccg ctggaactgg atcattgtgc
     1381 tggtgagcag cgacacctat ggccgcgaca atggccagct gcttggcgag cgcgtggccc
     1441 ggcgcgacat ctgcatcgcc ttccaggaga cgctgcccac actgcagccc aaccagaaca
     1501 tgacgtcaga ggagcgccag cgcctggtga ccattgtgga caagctgcag cagagcacag
     1561 cgcgcgtcgt ggtcgtgttc tcgcccgacc tgaccctgta ccacttcttc aatgaggtgc
     1621 tgcgccagaa cttcactggc gccgtgtgga tcgcctccga gtcctgggcc atcgacccgg
     1681 tcctgcacaa cctcacggag ctgcgccact tgggcacctt cctgggcatc accatccaga
     1741 gcgtgcccat cccgggcttc agtgagttcc gcgagtgggg cccacaggct gggccgccac
     1801 ccctcagcag gaccagccag agctatacct gcaaccagga gtgcgacaac tgcctgaacg
     1861 ccaccttgtc cttcaacacc attctcaggc tctctgggga gcgtgtcgtc tacagcgtgt
     1921 actctgcggt ctatgctgtg gcccatgccc tgcacagcct cctcggctgt gacaaaagca
     1981 cctgcaccaa gagggtggtc tacccctggc agctgcttga ggagatctgg aaggtcaact
     2041 tcactctcct ggaccaccaa atcttcttcg acccgcaagg ggacgtggct ctgcacttgg
     2101 agattgtcca gtggcaatgg gaccggagcc agaatccctt ccagagcgtc gcctcctact
     2161 accccctgca gcgacagctg aagaacatcc aagacatctc ctggcacacc atcaacaaca
     2221 cgatccctat gtccatgtgt tccaagaggt gccagtcagg gcaaaagaag aagcctgtgg
     2281 gcatccacgt ctgctgcttc gagtgcatcg actgccttcc cggcaccttc ctcaaccaca
     2341 ctgaagatga atatgaatgc caggcctgcc cgaataacga gtggtcctac cagagtgaga
     2401 cctcctgctt caagcggcag ctggtcttcc tggaatggca tgaggcaccc accatcgctg
     2461 tggccctgct ggccgccctg ggcttcctca gcaccctggc catcctggtg atattctgga
     2521 ggcacttcca gacacccata gttcgctcgg ctgggggccc catgtgcttc ctgatgctga
     2581 cactgctgct ggtggcatac atggtggtcc cggtgtacgt ggggccgccc aaggtctcca
     2641 cctgcctctg ccgccaggcc ctctttcccc tctgcttcac aatctgcatc tcctgtatcg
     2701 ccgtgcgttc tttccagatc gtctgcgcct tcaagatggc cagccgcttc ccacgcgcct
     2761 acagctactg ggtccgctac caggggccct acgtctctat ggcatttatc acggtactca
     2821 aaatggtcat tgtggtaatt ggcatgctgg ccacgggcct cagtcccacc acccgtactg
     2881 accccgatga ccccaagatc acaattgtct cctgtaaccc caactaccgc aacagcctgc
     2941 tgttcaacac cagcctggac ctgctgctct cagtggtggg tttcagcttc gcctacatgg
     3001 gcaaagagct gcccaccaac tacaacgagg ccaagttcat caccctcagc atgaccttct
     3061 atttcacctc atccgtctcc ctctgcacct tcatgtctgc ctacagcggg gtgctggtca
     3121 ccatcgtgga cctcttggtc actgtgctca acctcctggc catcagcctg ggctacttcg
     3181 gccccaagtg ctacatgatc ctcttctacc cggagcgcaa cacgcccgcc tacttcaaca
     3241 gcatgatcca gggctacacc atgaggaggg acggatccga ctacaaagac catgacggtg
     3301 attataaaga tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgata
     3361 tctcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     3421 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     3481 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     3541 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     3601 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     3661 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     3721 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     3781 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     3841 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     3901 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     3961 aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     4021 acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg
     4081 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     4141 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     4201 cgcagccaac gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag
     4261 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4321 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4381 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
     4441 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4501 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4561 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4621 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
     4681 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4741 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4801 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4861 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4921 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4981 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     5041 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     5101 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     5161 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     5221 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     5281 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     5341 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     5401 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     5461 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     5521 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     5581 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     5641 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     5701 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5761 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5821 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5881 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc
     5941 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     6001 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     6061 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     6121 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     6181 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     6241 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6301 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     6361 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6421 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6481 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6541 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6601 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6661 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6721 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6781 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6841 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6901 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6961 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7021 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7081 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7141 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7201 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7261 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7321 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7381 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7441 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7501 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7561 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7621 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7681 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7741 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7801 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7861 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7921 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7981 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     8041 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     8101 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8161 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8221 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8281 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8341 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8401 tgacgtcgac ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc
     8461 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     8521 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     8581 aatctgctta gg
//
