LOCUS       sequence-432278-        8216 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8216)
  AUTHORS   Xu XZ, Li HS, Guggino WB, Montell C
  TITLE     Coassembly of TRP and TRPL produces a distinct store-operated 
            conductance.
  JOURNAL   Cell. 1997 Jun 27. 89(7):1155-64.
  PUBMED    9215637
REFERENCE   2  (bases 1 to 8216)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 1997 
            Jun 27. 89(7):1155-64."
COMMENT     Sequence Label: TRPC1-FLAG
FEATURES             Location/Qualifiers
     source          1..8216
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      921..930
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             930..953
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3807..3824)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3813..4037
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4083..4511
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4170..4189)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4380..4401
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4520..4540)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4525..4854
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4705..4840
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4767..4786
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4921..5715
                     /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(4975..4994)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5585..5604
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5889..6010
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5926..5945)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5980..5999
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6059..6075)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6059..6075)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6072..6094)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6083..6099
                     /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(join(6107..6113,6114..6131,6132..6137))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6152..6173
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6290..6307)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6461..7049)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6541..6560)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7220..8011,8012..8080))
                     /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     7843..7862
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8081..8185)
                     /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 gtttaaacgg gccctctaga gccaccatgg actacaagga cgacgatgac aagcctgcag
      961 ggatggcggc cctgtacccg agcacggacc tctcgggcgc ctcctcctcc tccctgcctt
     1021 cctctccatc ctcttcctcg ccgaacgagg tgatggcgct gaaggatgtg cgggaggtga
     1081 aggaggagaa tacgctgaat gagaagcttt tcttgctggc gtgcgacaag ggtgactatt
     1141 atatggttaa aaagattttg gaggaaaaca gttcaggtga cttgaacata aattgcgtag
     1201 atgtgcttgg gagaaatgct gttaccataa ctattgaaaa cgaaaacttg gatatactgc
     1261 agcttctttt ggactacggt tgtcagaaac taatggaacg aattcagaat cctgagtatt
     1321 caacaactat ggatgttgca cctgtcattt tagctgctca tcgtaacaac tatgaaattc
     1381 ttacaatgct cttaaaacag gatgtatctc tacccaagcc ccatgcagtt ggctgtgaat
     1441 gcacattgtg ttctgcaaaa aacaaaaagg atagcctccg gcattccagg tttcgtcttg
     1501 atatatatcg atgtttggcc agtccagctc taataatgtt aacagaggag gatccaattc
     1561 tgagagcatt tgaacttagt gctgatttaa aagaactaag tcttgtggag gtggaattca
     1621 ggaatgatta tgaggaacta gcccggcaat gtaaaatgtt tgctaaggat ttacttgcac
     1681 aagcccggaa ttctcgtgaa ttggaagtta ttctaaacca tacgtctagt gacgagcctc
     1741 ttgacaaacg gggattatta gaagaaagaa tgaatttaag tcgtctaaaa cttgctatca
     1801 aatataacca gaaagagttt gtctcccagt ctaactgcca gcagttcctg aacactgttt
     1861 ggtttggaca gatgtcgggt taccgacgca agcccacctg taagaagata atgactgttt
     1921 tgacagtagg catcttttgg ccagttttgt cactttgtta tttgatagct cccaaatctc
     1981 agtttggcag aatcattcac acacctttta tgaaatttat cattcatgga gcatcatatt
     2041 tcacatttct gctgttgctt aatctatact ctcttgttta caatgaggat aagaaaaaca
     2101 caatggggcc agcccttgaa agaatagact atcttcttat tctgtggatt attgggatga
     2161 tttggtcaga cattaaaaga ctctggtatg aagggttgga agacttttta gaagaatctc
     2221 gtaatcaact cagttttgtc atgaattctc tttatttggc aacctttgcc ctcaaagtgg
     2281 ttgctcacaa caagtttcat gattttgctg atcggaagga ttgggatgca ttccatccta
     2341 cactggtggc agaagggctt tttgcatttg caaatgttct aagttatctt cgtctctttt
     2401 ttatgtatac aaccagctct atcttgggtc cattacagat ttcaatggga cagatgttac
     2461 aagattttgg aaaatttctt gggatgtttc ttcttgtttt gttttctttc acaattggac
     2521 tgacacaact gtatgataaa ggatatactt caaaggagca gaaggactgt gtaggcatct
     2581 tctgtgaaca gcaaagcaat gataccttcc attcgttcat tggcacctgc tttgctttgt
     2641 tctggtatat tttctcctta gcgcatgtgg caatctttgt cacaagattt agctatggag
     2701 aagaactgca gtcctttgtg ggagctgtca ttgttggtac atacaatgtc gtggttgtga
     2761 ttgtgcttac caaactgctg gtggcaatgc ttcataaaag ctttcagttg atagcaaatc
     2821 atgaagacaa agaatggaag tttgctcgag caaaattatg gcttagctac tttgatgaca
     2881 aatgtacgtt acctccacct ttcaacatca ttccctcacc aaagactatc tgctatatga
     2941 ttagtagcct cagtaagtgg atttgctctc atacatcaaa aggcaaggtc aaacggcaaa
     3001 acagtttaaa ggaatggaga aatttgaaac agaagagaga tgaaaactat caaaaagtga
     3061 tgtgctgcct agtgcatcgt tacttgactt ccatgagaca gaagatgcaa agtacagatc
     3121 aggcaactgt ggaaaatcta aacgaactgc gccaagatct gtcaaaattc cgaaatgaaa
     3181 taagggattt acttggcttt cggacttcta aatatgctat gttttatcca agaaattaac
     3241 cattttctaa atcatggagc gaataatttt caataacaga tccaaaagac tatattgcat
     3301 aacttgcaat gaaattaatg agatatatat tgaaataaag aattatgtaa aagccattct
     3361 ttaaaatatt tatagcataa atatatgtta tgtaaagtgt gtatatagaa ttagtttttt
     3421 aaaccttctg ttagtggctt tttgcagaag caaaacagat taagtagata gattttgtta
     3481 gcatgctgct tggttttctt acttagtgct ttaaaatgtt tttttttatg tttaagaggg
     3541 gcagttataa atggacacat tgcccagaat gttttgtaaa atgaagacca gcaaatgtag
     3601 gctgatctcc ttcacaggat acacttgaaa tatagaagtt atgttttaaa tatctctgtt
     3661 ttaggagttc acatatagtt cagcatttat tgtttaggag tataatttta ttttatctaa
     3721 aataatagtc tattttttct tttgtatttt gttataatct taaaattcat cgatggtacc
     3781 aagcttaagt ttaaaccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     3841 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     3901 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     3961 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     4021 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     4081 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4141 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4201 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4261 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4321 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4381 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4441 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4501 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4561 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4621 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4681 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4741 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     4801 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     4861 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     4921 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4981 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     5041 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     5101 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     5161 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     5221 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     5281 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     5341 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     5401 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     5461 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     5521 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     5581 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5641 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5701 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     5761 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5821 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5881 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5941 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6001 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     6061 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     6121 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     6181 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     6241 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     6301 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6361 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6421 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6481 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6541 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6601 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6661 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6721 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6781 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6841 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6901 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6961 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     7021 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7081 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7141 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7201 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7261 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7321 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7381 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7441 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7501 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7561 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7621 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7681 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7741 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7801 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7861 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7921 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7981 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8041 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     8101 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8161 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
