LOCUS       V018431                 8792 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018431
VERSION     V018431
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"
     CDS             671..3463
                     /label="TRPC6(NM_004621)"
                     /note="TRPC6(NM_004621)"
                     /gene="TRPC6"
     CDS             3479..3544
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3568..4156
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4222..4270
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4472..4900
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4914..5243
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5310..6104
                     /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    6280..6401
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6450..6466)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6474..6490
                     /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(6498..6528)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6543..6564
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6852..7440)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7611..8471)
                     /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(8472..8576)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgccacc atgagccaga gcccggcgtt cgggccccgg aggggcagtt ctccccgggg
      721 cgctgccgga gccgctgcgc ggcgcaacga gagccaggac tatctgctca tggactcgga
      781 gctgggagaa gacggctgcc cgcaagcccc gctgccttgc tacggctact acccctgctt
      841 ccggggatct gacaacagac tggctcaccg gcggcagaca gttctccgtg agaaggggag
      901 aaggttagct aatcgaggac cagcatacat gtttagtgat cgctccacaa gcctatctat
      961 agaggaggaa cgctttttgg atgcagctga atatggtaac atcccagtgg tgcggaagat
     1021 gttagaagaa tgccactcac tcaacgttaa ctgtgtggat tacatgggcc agaatgccct
     1081 acagttggca gtggccaatg agcatctgga aattacagaa cttcttctca agaaagaaaa
     1141 cctctctcga gttggggatg ctttgcttct agctattagt aaaggttatg ttcggattgt
     1201 ggaagcaatt ctcagtcatc cggcttttgc tgaaggcaag aggttagcaa ccagccctag
     1261 ccagtctgaa ctccagcaag atgattttta tgcctatgat gaagatggga cacggttctc
     1321 ccatgatgtg actccaatca ttctggctgc ccactgccag gaatatgaaa ttgtgcatac
     1381 cctcctgcgg aagggtgcta ggattgaacg gcctcatgat tatttctgca agtgcaatga
     1441 ctgcaaccag aaacagaagc atgactcgtt tagccactcc agatctagga ttaatgccta
     1501 taaaggcctg gcaagtccgg cttacctgtc attgtctagt gaagatccag tcatgacggc
     1561 tttagaactt agcaatgaac tggcagttct ggccaatatt gagaaagagt tcaagaatga
     1621 ctacaaaaaa ctgtcaatgc agtgcaaaga ctttgttgtt ggactccttg atctgtgcag
     1681 aaacactgaa gaagtcgagg ccattctgaa tggggatgtt gaaacgctcc agagtggtga
     1741 tcacggtcgc ccaaatctca gccgtttaaa acttgccatt aaatatgaag taaaaaaatt
     1801 tgtagctcat ccaaactgcc aacagcaact tctctccatt tggtatgaga atctttctgg
     1861 tttacgacag cagacaatgg cggtcaagtt ccttgtggtc cttgctgttg ccattggact
     1921 gcccttcctg gctctcattt actggtttgc tccatgcagc aagatgggga agataatgcg
     1981 tggaccattc atgaagtttg tagcacacgc agcctccttc accatttttc tgggactgct
     2041 agtcatgaat gcagctgaca gatttgaagg cacaaaactc cttcctaatg aaaccagcac
     2101 agataatgca aaacagctgt tcaggatgaa aacatcctgc ttctcatgga tggagatgct
     2161 cattatatcc tgggtaatag gcatgatatg ggctgaatgt aaagaaatct ggactcaggg
     2221 ccccaaggaa tatttgtttg agttgtggaa catgcttgat tttggtatgt tagcaatttt
     2281 cgcagcatca ttcattgcga gattcatggc attttggcat gcttccaaag cccagagcat
     2341 cattgacgca aatgatactt tgaaggactt gacgaaagta acattgggag acaatgtgaa
     2401 atactacaat ttggccagga taaagtggga cccctctgat cctcaaataa tatctgaagg
     2461 tctttatgca attgctgtag ttttaagttt ctctaggata gcttatattt taccagcaaa
     2521 tgaaagcttt ggacctctgc agatatcact tggaagaaca gtcaaagaca tcttcaagtt
     2581 catggtcata ttcattatgg tgtttgtggc ctttatgatt ggaatgttca atctctactc
     2641 ctactacatt ggtgcaaaac aaaatgaagc cttcacaaca gttgaagaga gttttaagac
     2701 actgttctgg gctatatttg gactttctga agtgaaatca gtggtcatca actataacca
     2761 caaattcatt gaaaacattg gttacgttct ttatggagtc tataatgtta cgatggtcat
     2821 tgttttgcta aatatgttaa ttgccatgat caacagttca ttccaggaaa ttgaggatga
     2881 cgctgatgtg gagtggaaat ttgcaagggc caaactctgg ttttcctact ttgaggaggg
     2941 cagaacactt cctgtaccct tcaatctggt gccgagtcca aagtccctgt tttatctctt
     3001 actgaagctt aaaaaatgga tttctgagct gttccagggc cataaaaaag gtttccagga
     3061 agatgcagag atgaacaaga taaatgaaga aaagaaactt ggaattttag gaagtcatga
     3121 agacctttca aaattatcac ttgacaaaaa acaggttggg cacaataaac aaccaagtat
     3181 aaggagctca gaagatttcc atctaaatag tttcaataat cctccaagac aatatcagaa
     3241 aataatgaaa aggctcatta aaagatatgt actgcaggcc cagatagata aggagagtga
     3301 tgaagtgaac gaaggggaac tgaaggaaat taagcaggac atctcaagtc tccgctatga
     3361 actccttgaa gaaaaatctc agaatacaga agacctagca gaacttatta gagaacttgg
     3421 agagaaatta tccatggaac caaatcaaga ggaaaccaat agactgcggc cgctcgagga
     3481 ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga
     3541 caagtagtga gggcccgata tctcgacaat caacctctgg attacaaaat ttgtgaaaga
     3601 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     3661 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     3721 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     3781 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     3841 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     3901 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     3961 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     4021 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     4081 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     4141 tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag gagacaatac
     4201 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac gggtgttggg
     4261 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     4321 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     4381 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcagat
     4441 ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     4501 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4561 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4621 aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4681 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4741 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4801 aaccctatct cggtctattc ttttgattta taagggattt tggggatttc ggcctattgg
     4861 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     4921 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4981 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5041 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     5101 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     5161 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     5221 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     5281 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     5341 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     5401 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     5461 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     5521 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     5581 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     5641 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     5701 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5761 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     5821 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5881 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5941 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     6001 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     6061 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     6121 cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     6181 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     6241 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     6301 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     6361 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     6421 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     6481 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     6541 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     6601 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     6661 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     6721 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6781 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6841 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6901 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6961 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     7021 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     7081 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     7141 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     7201 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     7261 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     7321 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7381 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7441 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7501 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     7561 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     7621 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     7681 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7741 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7801 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7861 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7921 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7981 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     8041 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     8101 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     8161 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     8221 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     8281 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     8341 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     8401 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     8461 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     8521 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     8581 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     8641 cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     8701 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     8761 tgaccgacaa ttgcatgaag aatctgctta gg
//
