LOCUS       sequence-210333-        7110 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  GsX chimera with G-alpha-i C-terminus (Pertussis toxin sensitive).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7110)
  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 7110)
  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 Gsi (Cys)
FEATURES             Location/Qualifiers
     source          1..7110
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          88..1104
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1028..1050
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1112..1131
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1174..1183
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2487..2506)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2552..2607
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2606..2625)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     protein_bind    2971..3004
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    3017..3138
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3054..3073)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3108..3127
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3157..3177)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3157..3175)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3196..3212)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3196..3212)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3209..3231)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3220..3236
                     /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(3244..3250,3251..3268,3269..3274))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3289..3310
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3427..3444)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3598..4186)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3678..3697)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4357..5148,5149..5217))
                     /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     4980..4999
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5218..5322)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    5395..5443
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    5655..5688
                     /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             5693..6292
                     /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(5693..5712)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6189..6209
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(6428..6445)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6434..6545
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        6690..6993
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 aatcagagcg gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc
       61 tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc
      121 cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag
      181 gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg
      241 cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc
      301 ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc
      361 tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc
      421 agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag
      481 gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg
      541 tcggtcgggc tgcaaccccc cctgcacccc cctccccgag ttgctgagca cggcccggct
      601 tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg
      661 cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg ccggggaggg ctcgggggag
      721 gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg cggcgagccg cagccattgc
      781 cttttatggt aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag
      841 ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc
      901 gccggcagga aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt
      961 ctccctctcc agcctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc
     1021 agggcggggt tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt
     1081 catgccttct tctttttcct acagctcctg ggcaacgtgc tggttattgt gctgtctcat
     1141 cattttggca aagaattgct cgaccaccgg cgcgccacca tgggctgcct cgggaacagt
     1201 aagaccgagg accagcgcaa cgaggagaag gcgcagcgtg aggccaacaa aaagatcgag
     1261 aagcagctgc agaaggacaa gcaggtctac cgggccacgc accgcctgct gctgctgggt
     1321 gctggagaat ctggtaaaag caccattgtg aagcagatga ggatcctgca tgttaatggg
     1381 tttaatggag agggcggcga agaggacccg caggctgcaa ggagcaacag cgatggtgag
     1441 aaggcaacca aagtgcagga catcaaaaac aacctgaaag aggcgattga aaccattgtg
     1501 gccgccatga gcaacctggt gccccccgtg gagctggcca accccgagaa ccagttcaga
     1561 gtggactaca ttctgagtgt gatgaacgtg cctgactttg acttccctcc cgaattctat
     1621 gagcatgcca aggctctgtg ggaggatgaa ggagtgcgtg cctgctacga acgctccaac
     1681 gagtaccagc tgattgactg tgcccagtac ttcctggaca agatcgacgt gatcaagcag
     1741 gctgactatg tgccgagcga tcaggacctg cttcgctgcc gtgtcctgac ttctggaatc
     1801 tttgagacca agttccaggt ggacaaagtc aacttccaca tgtttgacgt gggtggccag
     1861 cgcgatgaac gccgcaagtg gatccagtgc ttcaacgatg tgactgccat catcttcgtg
     1921 gtggccagca gcagctacaa catggtcatc cgggaggaca accagaccaa ccgcctgcag
     1981 gaggctctga acctcttcaa gagcatctgg aacaacagat ggctgcgcac catctctgtg
     2041 atcctgttcc tcaacaagca agatctgctc gctgagaaag tccttgctgg gaaatcgaag
     2101 attgaggact actttccaga atttgctcgc tacactactc ctgaggatgc tactcccgag
     2161 cccggagagg acccacgcgt gacccgggcc aagtacttca ttcgagatga gtttctgagg
     2221 atcagcactg ccagtggaga tgggcgtcac tactgctacc ctcatttcac ctgcgctgtg
     2281 gacactgaga acatccgccg tgtgttcaac gactgccgtg atgtcatcat aaaaaataat
     2341 ctaaaagatt gtggtctctt ttgaaccggt ctgcagccta ggcttaagcg ctcgaggcgg
     2401 ccgcaattca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat
     2461 gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc
     2521 atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata
     2581 gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta
     2641 aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc
     2701 atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat
     2761 tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg
     2821 ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc
     2881 ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac
     2941 ctgcaccgtc gagctagcat atggatccat ataacttcgt ataatgtatg ctatacgaag
     3001 ttattacgat accgtcaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     3061 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     3121 actcatcaat gtatcttagg tacccagctt ttgttccctt tagtgagggt taattgcgcg
     3181 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     3241 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     3301 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3361 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3421 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3481 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3541 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3601 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3661 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3721 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3781 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3841 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3901 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3961 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4021 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4081 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4141 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4201 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4261 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4321 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4381 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4441 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4501 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4561 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4621 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4681 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4741 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4801 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4861 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4921 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4981 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5041 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5101 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5161 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5221 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5281 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     5341 gccaagctta ataaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     5401 taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg
     5461 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     5521 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     5581 aacgtcgggg cggcaggccc tgccatagcc actggccccg tgggttaggg acggggtccc
     5641 ccattcgaga ataaataact tcgtataaag tatcctatac gaagttatca ccatgaccga
     5701 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct
     5761 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat
     5821 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     5881 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     5941 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     6001 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     6061 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     6121 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     6181 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     6241 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacgcccgcc
     6301 ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catggctccg accgaagcca
     6361 cccggggcgg ccccgccgac cccgcacccg cccccgaggc ccaccgactc tagagctcgc
     6421 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     6481 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     6541 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     6601 aagggggagg attgggaaga caatagcact agttattaat agtaatcaat tacggggtca
     6661 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     6721 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     6781 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     6841 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     6901 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     6961 tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc
     7021 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta
     7081 ttttgtgcag cgatgggggc gggggggggg
//
