LOCUS       Exported                8897 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    htrpC6-YFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8897)
  AUTHORS   Kwon Y, Hofmann T, Montell C
  TITLE     Integration of phosphoinositide- and calmodulin-mediated regulation 
            of TRPC6.
  JOURNAL   Mol Cell. 2007 Feb 23. 25(4):491-503.
  PUBMED    17317623
REFERENCE   2  (bases 1 to 8897)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8897
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3720..4436
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3762..3783)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4023..4042)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4370..4391
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(4452..4470)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4453..4470)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4490..4507)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4496..4720
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4766..5194
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4853..4872)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5063..5084
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5203..5223)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5208..5537
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5388..5523
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5450..5469
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5604..6398
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5658..5677)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6268..6287
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6572..6693
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6609..6628)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6663..6682
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6742..6758)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6742..6758)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6755..6777)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6766..6782
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(6790..6820)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6835..6856
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6973..6990)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7144..7732)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7224..7243)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7903..8763)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8526..8545
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8764..8868)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagcttggta
      901 ccgagctcgg atccaccgcc atgagccaga gcccggcgtt cgggccccgg aggggcagtt
      961 ctccccgggg cgctgccgga gccgctgcgc ggcgcaacga gagccaggac tatctgctca
     1021 tggactcgga gctgggagaa gacggctgcc cgcaagcccc gctgccttgc tacggctact
     1081 acccctgctt ccggggatct gacaacagac tggctcaccg gcggcagaca gttctccgtg
     1141 agaaggggag aaggttagct aatcgaggac cagcatacat gtttagtgat cgctccacaa
     1201 gcctatctat agaggaggaa cgctttttgg atgcagctga atatggtaac atcccagtgg
     1261 tgcggaagat gttagaagaa tgccactcac tcaacgttaa ctgtgtggat tacatgggcc
     1321 agaatgccct acagttggca gtggccaatg agcatctgga aattacagaa cttcttctca
     1381 agaaagaaaa cctctctcga gttggggatg ctttgcttct agctattagt aaaggttatg
     1441 ttcggattgt ggaagcaatt ctcagtcatc cggcttttgc tgaaggcaag aggttagcaa
     1501 ccagccctag ccagtctgaa ctccagcaag atgattttta tgcctatgat gaagatggga
     1561 cacggttctc ccatgatgtg actccaatca ttctggctgc ccactgccag gaatatgaaa
     1621 ttgtgcatac cctcctgcgg aagggtgcta ggattgaacg gcctcatgat tatttctgca
     1681 agtgcaatga ctgcaaccag aaacagaagc atgactcgtt tagccactcc agatctagga
     1741 ttaatgccta taaaggcctg gcaagtccgg cttacctgtc attgtctagt gaagatccag
     1801 tcatgacggc tttagaactt agcaatgaac tggcagttct ggccaatatt gagaaagagt
     1861 tcaagaatga ctacaaaaaa ctgtcaatgc agtgcaaaga ctttgttgtc ggactccttg
     1921 atctgtgcag aaacactgaa gaagtcgagg ccattctgaa tggggatgtt gaaacgctcc
     1981 agagtggtga tcacggtcgc ccaaatctca gccgtttaaa acttgccatt aaatatgaag
     2041 taaaaaaatt tgtagctcat ccaaactgcc aacagcaact tctctccatt tggtatgaga
     2101 atctttctgg tttacgacag cagacaatgg cggtcaagtt ccttgtggtc cttgctgttg
     2161 ccattggact gcccttcctg gctctcattt actggtttgc tccatgcagc aagatgggga
     2221 agataatgcg tggaccattc atgaagtttg tagcacacgc agcctccttc accatttttc
     2281 tgggactgct agtcatgaat gcagctgaca gatttgaagg cacaaaactc cttcctaatg
     2341 aaaccagcac agataatgca aaacagctgt tcaggatgaa aacatcctgc ttctcatgga
     2401 tggagatgct cattatatcc tgggtaatag gcatgatatg ggctgaatgt aaagaaatct
     2461 ggactcaggg ccccaaggaa tatttgtttg agttgtggaa catgcttgat tttggtatgt
     2521 tagcaatttt cgcagcatca ttcattgcga gattcatggc attttggcat gcttccaaag
     2581 cccagagcat cattgacgca aacgatactt tgaaggactt gacgaaagta acattgggag
     2641 acaatgtgaa atactacaat ttggccagga taaagtggga cccctctgat cctcaaataa
     2701 tatctgaagg tctttatgca attgctgtag ttttaagttt ctctaggata gcttatattt
     2761 taccagcaaa tgaaagcttt ggacctctgc agatatcact tggaagaaca gtcaaagaca
     2821 tcttcaagtt catggtcata ttcattatgg tgtttgtggc ctttatgatt ggaatgttca
     2881 atctctactc ctactacatt ggtgcaaaac aaaatgaagc cttcacaaca gttgaagaga
     2941 gttttaagac actgttctgg gctatatttg gactttctga agtgaaatca gtggtcatca
     3001 actataacca caaattcatt gaaaacattg gttacgttct ttatggagtc tataatgtta
     3061 cgatggtcat tgttttgcta aatatgttaa ttgccatgat caacagttca ttccaggaaa
     3121 ttgaggatga cgctgatgtg gagtggaaat ttgcaagggc caaactctgg ttttcctact
     3181 ttgaggaggg cagaacactt cctgtaccct tcaatctggt gccgagtcca aagtccctgt
     3241 tttatctctt actgaagctt aaaaaatgga tttctgagct gttccagggc cataaaaaag
     3301 gtttccagga agatgcagag atgaacaaga taaatgaaga aaagaaactt ggaattttag
     3361 gaagtcatga agacctttca aaattatcac ttgacaaaaa acaggttggg cacaataaac
     3421 aaccaagtat aaggagctca gaagatttcc atctaaatag tttcaataat cctccaagac
     3481 aatatcagaa aataatgaaa aggctcatta aaagatatgt actgcaggcc cagatagata
     3541 aggagagtga tgaagtgaac gaaggggaac tgaaggaaat taagcaggac atctcaagtc
     3601 tccgctatga actccttgaa gaaaaatctc agaatacaga agacctagca gaacttatta
     3661 gagaacttgg agagaaatta tccatggaac caaatcaaga ggaaaccaat agagaattcg
     3721 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     3781 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     3841 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     3901 tgaccacctt cggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc
     3961 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     4021 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     4081 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     4141 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     4201 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     4261 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     4321 tgagctacca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     4381 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaatcta
     4441 gagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg
     4501 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     4561 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     4621 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     4681 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     4741 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4801 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4861 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4921 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4981 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     5041 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     5101 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     5161 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     5221 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     5281 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     5341 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     5401 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     5461 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     5521 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     5581 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     5641 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     5701 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     5761 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     5821 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     5881 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     5941 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     6001 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     6061 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     6121 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     6181 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     6241 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     6301 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     6361 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     6421 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     6481 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     6541 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     6601 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     6661 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     6721 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6781 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6841 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6901 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6961 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     7021 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     7081 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     7141 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     7201 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     7261 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     7321 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     7381 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     7441 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7501 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7561 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     7621 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7681 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7741 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7801 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7861 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7921 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7981 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     8041 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     8101 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     8161 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     8221 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     8281 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     8341 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     8401 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     8461 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     8521 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     8581 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     8641 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8701 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8761 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8821 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8881 aaaagtgcca cctgacg
//
