LOCUS       sequence-210353-        7100 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  GsX chimera with G-alpha-o C-terminus (Pertussis toxin insensitive 
            mutant).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7100)
  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 7100)
  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 Gso (Ser)
FEATURES             Location/Qualifiers
     source          1..7100
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          123..1139
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1063..1085
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1147..1166
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1209..1218
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2522..2541)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2587..2642
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2641..2660)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     protein_bind    3006..3039
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    3052..3173
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3089..3108)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3143..3162
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3192..3212)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3192..3210)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3231..3247)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3231..3247)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3244..3266)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3255..3271
                     /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(3279..3285,3286..3303,3304..3309))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3324..3345
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3462..3479)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3633..4221)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3713..3732)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4392..5183,5184..5252))
                     /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     5015..5034
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5253..5357)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    5430..5478
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    5690..5723
                     /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             5728..6327
                     /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(5728..5747)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6224..6244
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(6463..6480)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6469..6580
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        6725..7028
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
       61 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      121 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      181 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      241 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      301 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      361 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      421 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
      481 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
      541 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
      601 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
      661 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
      721 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
      781 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
      841 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
      901 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
      961 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1021 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1081 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1141 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttgctcgacc
     1201 accggcgcgc caccatgggc tgcctcggga acagtaagac cgaggaccag cgcaacgagg
     1261 agaaggcgca gcgtgaggcc aacaaaaaga tcgagaagca gctgcagaag gacaagcagg
     1321 tctaccgggc cacgcaccgc ctgctgctgc tgggtgctgg agaatctggt aaaagcacca
     1381 ttgtgaagca gatgaggatc ctgcatgtta atgggtttaa tggagagggc ggcgaagagg
     1441 acccgcaggc tgcaaggagc aacagcgatg gtgagaaggc aaccaaagtg caggacatca
     1501 aaaacaacct gaaagaggcg attgaaacca ttgtggccgc catgagcaac ctggtgcccc
     1561 ccgtggagct ggccaacccc gagaaccagt tcagagtgga ctacattctg agtgtgatga
     1621 acgtgcctga ctttgacttc cctcccgaat tctatgagca tgccaaggct ctgtgggagg
     1681 atgaaggagt gcgtgcctgc tacgaacgct ccaacgagta ccagctgatt gactgtgccc
     1741 agtacttcct ggacaagatc gacgtgatca agcaggctga ctatgtgccg agcgatcagg
     1801 acctgcttcg ctgccgtgtc ctgacttctg gaatctttga gaccaagttc caggtggaca
     1861 aagtcaactt ccacatgttt gacgtgggtg gccagcgcga tgaacgccgc aagtggatcc
     1921 agtgcttcaa cgatgtgact gccatcatct tcgtggtggc cagcagcagc tacaacatgg
     1981 tcatccggga ggacaaccag accaaccgcc tgcaggaggc tctgaacctc ttcaagagca
     2041 tctggaacaa cagatggctg cgcaccatct ctgtgatcct gttcctcaac aagcaagatc
     2101 tgctcgctga gaaagtcctt gctgggaaat cgaagattga ggactacttt ccagaatttg
     2161 ctcgctacac tactcctgag gatgctactc ccgagcccgg agaggaccca cgcgtgaccc
     2221 gggccaagta cttcattcga gatgagtttc tgaggatcag cactgccagt ggagatgggc
     2281 gtcactactg ctaccctcat ttcacctgcg ctgtggacac tgagaacatc cgccgtgtgt
     2341 tcaacgactg ccgtgacatc atcatcgcca aaaacctgcg gggctctgga ctctactgaa
     2401 ccggtctgca gcctaggctt aagcgctcga ggcggccgca attcactcct caggtgcagg
     2461 ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag
     2521 atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt
     2581 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct
     2641 cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt
     2701 agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag
     2761 gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga
     2821 cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa
     2881 aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca
     2941 tagctgtccc tcttctctta tggagatccc tcgacctgca ccgtcgagct agcatatgga
     3001 tccatataac ttcgtataat gtatgctata cgaagttatt acgataccgt caacttgttt
     3061 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3121 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaggtaccc
     3181 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
     3241 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     3301 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     3361 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attgatgaat cggccaacgc
     3421 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     3481 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3541 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3601 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3661 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3721 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3781 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3841 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3901 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3961 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4021 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4081 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4141 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4201 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4261 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4321 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4381 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4441 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4501 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4561 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4621 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4681 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4741 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4801 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4861 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4921 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4981 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5041 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5101 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5161 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5221 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5281 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5341 caaatagggg ttccgcgcac atttccccga aaagtgccaa gcttaataaa cacggaagga
     5401 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     5461 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     5521 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     5581 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     5641 tagccactgg ccccgtgggt tagggacggg gtcccccatt cgagaataaa taacttcgta
     5701 taaagtatcc tatacgaagt tatcaccatg accgagtaca agcccacggt gcgcctcgcc
     5761 acccgcgacg acgtcccccg ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     5821 gccacgcgcc acaccgtcga cccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     5881 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     5941 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     6001 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
     6061 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
     6121 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
     6181 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     6241 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     6301 tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa
     6361 aggagcgcac gaccccatgg ctccgaccga agccacccgg ggcggccccg ccgaccccgc
     6421 acccgccccc gaggcccacc gactctagag ctcgctgatc agcctcgact gtgccttcta
     6481 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     6541 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     6601 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     6661 gcactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     6721 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     6781 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     6841 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     6901 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     6961 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     7021 ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc
     7081 cacccccaat tttgtattta
//
