LOCUS       V021832                 6652 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021832
VERSION     V021832
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"
     regulatory      901..910
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             910..933
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             943..2172
                     /label="Gprc5b(NM_022420)"
                     /note="Gprc5b(NM_022420)"
                     /gene="Gprc5b"
     polyA_signal    2252..2476
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2522..2950
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2964..3293
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3360..4154
                     /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    4328..4449
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4498..4514)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4522..4538
                     /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(4546..4576)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4591..4612
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4900..5485)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5656..6516)
                     /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(6517..6621)
                     /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 gccaccatgg attacaagga tgacgacgat aagactaagc ttatgttcct ggtgttagag
      961 agaaagatga gaacccatca agtgtttccc ttgcccctgc tcctggtgat tgcctccgtg
     1021 gcttcagaga acgccagcac gtcccggggc tgtggactgg accttcttcc tcagtacgtg
     1081 tccctgtgcg acctggacgc catctggggc atcgtggtgg aggcagtggc cggggcgggg
     1141 gccctgatca cactgcttct gatgctcatt ctcctagtga gactaccctt catcaaggac
     1201 aaggaaagga agcggcctgt gtgcctccat ttcctcttcc tgctggggac cctgggcctc
     1261 tttggcctga cgtttgcctt catcatccag atggacgaga caatctgctc catccgacgc
     1321 ttcctctggg gtgtcctctt cgcgctctgc ttttcctgcc tgctgagcca ggcgtggcgg
     1381 gtgcggaggc tggtgcgcca gggcacaagc ccggccagct ggcagctggt gagcctggca
     1441 ctgtgcctga tgctggtgca ggtcatcatt gccactgagt ggctggtgct gactgtgctg
     1501 cgtgacacga agccagcctg cgcctacgag cccatggatt ttgtgatggc gctcatctac
     1561 gacatggtgc tgctggccat caccctggcc cagtccctct tcacgctgtg tggcaagttc
     1621 aaacggtgga aggtgaacgg agccttcatc ctcgtcacta ccttcctctc tgcgctcatc
     1681 tgggtggtct ggatgaccat gtacctcttc ggcaactcat taattaagca gggagatgcc
     1741 tggagcgacc ccaccttggc cattacactg gcagccagcg gatgggtctt tgtcatcttc
     1801 catgccattc cggagatcca ctacaccctt ctcccacccc tgcaggagaa cccacccaac
     1861 tacttcgaca cttcacagcc caggatgcgg gagacagcat ttgatgagga aatgcacctg
     1921 cctcgggcct acatggagaa caaggccttc tcaatggatg aacataacgc agctctccga
     1981 tcagcagtgg gcttttccaa tggaagcttg gagcaaagat ctagcagcct gggaaagaag
     2041 cccagcagct tgggaaaccg gcccagtgca ccgttcagaa gcaatgtgta tcagccgacc
     2101 gagatggccg tcgtgctcaa tggagggact atcccaactg ctccgccatc tcacacggga
     2161 agacaccatt ggtgagaatt ctgcagatat ccagcacagt ggcggccgct cgagtctaga
     2221 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     2281 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2341 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     2401 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     2461 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     2521 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2581 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     2641 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     2701 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     2761 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2821 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     2881 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     2941 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3001 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3061 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3121 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3181 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     3241 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     3301 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     3361 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     3421 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     3481 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     3541 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     3601 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     3661 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     3721 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     3781 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     3841 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     3901 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     3961 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4021 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4081 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4141 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     4201 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4261 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4321 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4381 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4441 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     4501 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4561 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4621 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4681 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4741 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4801 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4861 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4921 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4981 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5041 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5101 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5161 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5221 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5281 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5341 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5401 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat
     5461 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5521 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5581 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5641 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5701 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5761 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5821 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5881 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5941 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6001 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6061 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6121 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6181 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6241 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6301 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6361 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6421 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6481 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6541 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6601 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
