LOCUS       sequence-210403-        7109 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  GsX chimera with G-alpha-t C-terminus (Pertussis toxin sensitive).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7109)
  AUTHORS   Ballister ER, Rodgers J, Martial F, Lucas RJ
  TITLE     A live cell assay of GPCR coupling allows identification of 
            optogenetic tools for controlling Go and Gi signaling.
  JOURNAL   BMC Biol. 2018 Jan 16;16(1):10. doi: 10.1186/s12915-017-0475-2.
  PUBMED    29338718
REFERENCE   2  (bases 1 to 7109)
  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: "BMC Biol.";
            date: "2018-01-16"; volume: "16(1):10. doi"; pages: " 
            10.1186/s12915-017-0475-2"
COMMENT     Sequence Label: pEM705 Gst (Cys)
FEATURES             Location/Qualifiers
     source          1..7109
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          113..1129
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1053..1075
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1137..1156
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1199..1208
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2486..2505)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2551..2606
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2605..2624)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     protein_bind    2970..3003
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    3016..3137
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3053..3072)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3107..3126
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3156..3176)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3156..3174)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3195..3211)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3195..3211)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3208..3230)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3219..3235
                     /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(3243..3249,3250..3267,3268..3273))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3288..3309
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3426..3443)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3597..4185)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3677..3696)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4356..5147,5148..5216))
                     /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     4979..4998
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5217..5321)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    5394..5442
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    5654..5687
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     CDS             5692..6291
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5692..5711)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6188..6208
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(6427..6444)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6433..6544
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        6689..6992
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat
       61 ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc
      121 tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct
      181 ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg
      241 taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg aaagccttga
      301 ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt gcgtgtgtgt
      361 gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc gctgcgggcg
      421 cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg ggggcggtgc
      481 cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg tgtgcgtggg
      541 ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc acccccctcc
      601 ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc gtggcgcggg
      661 gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg cggggccgcc
      721 tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg gcggctgtcg
      781 aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg cgcagggact
      841 tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca ccccctctag
      901 cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg agggccttcg
      961 tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc cgcgggggga
     1021 cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg
     1081 ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc tcctgggcaa
     1141 cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttgctcgacc accggcgcgc
     1201 caccatgggc tgcctcggga acagtaagac cgaggaccag cgcaacgagg agaaggcgca
     1261 gcgtgaggcc aacaaaaaga tcgagaagca gctgcagaag gacaagcagg tctaccgggc
     1321 cacgcaccgc ctgctgctgc tgggtgctgg agaatctggt aaaagcacca ttgtgaagca
     1381 gatgaggatc ctgcatgtta atgggtttaa tggagagggc ggcgaagagg acccgcaggc
     1441 tgcaaggagc aacagcgatg gtgagaaggc aaccaaagtg caggacatca aaaacaacct
     1501 gaaagaggcg attgaaacca ttgtggccgc catgagcaac ctggtgcccc ccgtggagct
     1561 ggccaacccc gagaaccagt tcagagtgga ctacattctg agtgtgatga acgtgcctga
     1621 ctttgacttc cctcccgaat tctatgagca tgccaaggct ctgtgggagg atgaaggagt
     1681 gcgtgcctgc tacgaacgct ccaacgagta ccagctgatt gactgtgccc agtacttcct
     1741 ggacaagatc gacgtgatca agcaggctga ctatgtgccg agcgatcagg acctgcttcg
     1801 ctgccgtgtc ctgacttctg gaatctttga gaccaagttc caggtggaca aagtcaactt
     1861 ccacatgttt gacgtgggtg gccagcgcga tgaacgccgc aagtggatcc agtgcttcaa
     1921 cgatgtgact gccatcatct tcgtggtggc cagcagcagc tacaacatgg tcatccggga
     1981 ggacaaccag accaaccgcc tgcaggaggc tctgaacctc ttcaagagca tctggaacaa
     2041 cagatggctg cgcaccatct ctgtgatcct gttcctcaac aagcaagatc tgctcgctga
     2101 gaaagtcctt gctgggaaat cgaagattga ggactacttt ccagaatttg ctcgctacac
     2161 tactcctgag gatgctactc ccgagcccgg agaggaccca cgcgtgaccc gggccaagta
     2221 cttcattcga gatgagtttc tgaggatcag cactgccagt ggagatgggc gtcactactg
     2281 ctaccctcat ttcacctgcg ctgtggacac tgagaacatc cgccgtgtgt tcaacgactg
     2341 ccgtgacatc atcatcaagg agaacctcaa agactgtggc ctcttctgac tcgaggcggc
     2401 cgcaattcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg
     2461 ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca
     2521 tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag
     2581 tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa
     2641 aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca
     2701 tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt
     2761 ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt
     2821 tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc
     2881 tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga tccctcgacc
     2941 tgcaccgtcg agctagcata tggatccata taacttcgta taatgtatgc tatacgaagt
     3001 tattacgata ccgtcaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3061 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3121 ctcatcaatg tatcttaggt acccagcttt tgttcccttt agtgagggtt aattgcgcgc
     3181 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     3241 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     3301 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     3361 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3421 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3481 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3541 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3601 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3661 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3721 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3781 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3841 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3901 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3961 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4021 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4081 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4141 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4201 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4261 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4321 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4381 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4441 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4501 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4561 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4621 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     4681 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4741 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4801 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4861 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4921 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4981 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5041 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5101 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5161 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5221 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5281 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5341 ccaagcttaa taaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     5401 aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     5461 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     5521 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     5581 acgtcggggc ggcaggccct gccatagcca ctggccccgt gggttaggga cggggtcccc
     5641 cattcgagaa taaataactt cgtataaagt atcctatacg aagttatcac catgaccgag
     5701 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt acgcaccctc
     5761 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgacccgga ccgccacatc
     5821 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     5881 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     5941 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     6001 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     6061 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     6121 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     6181 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     6241 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg acgcccgccc
     6301 cacgacccgc agcgcccgac cgaaaggagc gcacgacccc atggctccga ccgaagccac
     6361 ccggggcggc cccgccgacc ccgcacccgc ccccgaggcc caccgactct agagctcgct
     6421 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     6481 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     6541 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     6601 agggggagga ttgggaagac aatagcacta gttattaata gtaatcaatt acggggtcat
     6661 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     6721 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     6781 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     6841 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     6901 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     6961 acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg ttctgcttca
     7021 ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat
     7081 tttgtgcagc gatgggggcg ggggggggg
//
