LOCUS       sequence-210347-        7096 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  GsX chimera with G-alpha-12 C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7096)
  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 7096)
  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 Gs12
FEATURES             Location/Qualifiers
     source          1..7096
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          102..1118
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1042..1064
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1126..1145
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1188..1197
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2475..2494)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2540..2595
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2594..2613)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     protein_bind    2959..2992
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    3005..3126
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3042..3061)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3096..3115
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3145..3165)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3145..3163)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3184..3200)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3184..3200)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3197..3219)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3208..3224
                     /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(3232..3238,3239..3256,3257..3262))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3277..3298
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3415..3432)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3586..4174)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3666..3685)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4345..5136,5137..5205))
                     /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     4968..4987
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5206..5310)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    5383..5431
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    5643..5676
                     /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             5681..6280
                     /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(5681..5700)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6177..6197
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(6416..6433)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6422..6533
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        6678..6981
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc
       61 ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc
      121 ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg
      181 cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagcgct
      241 tggtttaatg acggcttgtt tcttttctgt ggctgcgtga aagccttgag gggctccggg
      301 agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga
      361 gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc ggcgcggggc
      421 tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc ccgcggtgcg
      481 gggggggctg cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg gggtgagcag
      541 ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca cccccctccc cgagttgctg
      601 agcacggccc ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg ctcgccgtgc
      661 cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct cgggccgggg
      721 agggctcggg ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga ggcgcggcga
      781 gccgcagcca ttgcctttta tggtaatcgt gcgagagggc gcagggactt cctttgtccc
      841 aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc gggcgcgggg
      901 cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc
      961 gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac ggctgccttc
     1021 gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagagcct
     1081 ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta
     1141 ttgtgctgtc tcatcatttt ggcaaagaat tgctcgacca ccggcgcgcc accatgggct
     1201 gcctcgggaa cagtaagacc gaggaccagc gcaacgagga gaaggcgcag cgtgaggcca
     1261 acaaaaagat cgagaagcag ctgcagaagg acaagcaggt ctaccgggcc acgcaccgcc
     1321 tgctgctgct gggtgctgga gaatctggta aaagcaccat tgtgaagcag atgaggatcc
     1381 tgcatgttaa tgggtttaat ggagagggcg gcgaagagga cccgcaggct gcaaggagca
     1441 acagcgatgg tgagaaggca accaaagtgc aggacatcaa aaacaacctg aaagaggcga
     1501 ttgaaaccat tgtggccgcc atgagcaacc tggtgccccc cgtggagctg gccaaccccg
     1561 agaaccagtt cagagtggac tacattctga gtgtgatgaa cgtgcctgac tttgacttcc
     1621 ctcccgaatt ctatgagcat gccaaggctc tgtgggagga tgaaggagtg cgtgcctgct
     1681 acgaacgctc caacgagtac cagctgattg actgtgccca gtacttcctg gacaagatcg
     1741 acgtgatcaa gcaggctgac tatgtgccga gcgatcagga cctgcttcgc tgccgtgtcc
     1801 tgacttctgg aatctttgag accaagttcc aggtggacaa agtcaacttc cacatgtttg
     1861 acgtgggtgg ccagcgcgat gaacgccgca agtggatcca gtgcttcaac gatgtgactg
     1921 ccatcatctt cgtggtggcc agcagcagct acaacatggt catccgggag gacaaccaga
     1981 ccaaccgcct gcaggaggct ctgaacctct tcaagagcat ctggaacaac agatggctgc
     2041 gcaccatctc tgtgatcctg ttcctcaaca agcaagatct gctcgctgag aaagtccttg
     2101 ctgggaaatc gaagattgag gactactttc cagaatttgc tcgctacact actcctgagg
     2161 atgctactcc cgagcccgga gaggacccac gcgtgacccg ggccaagtac ttcattcgag
     2221 atgagtttct gaggatcagc actgccagtg gagatgggcg tcactactgc taccctcatt
     2281 tcacctgcgc tgtggacact gagaacatcc gccgtgtgtt caacgactgc cgtgacacca
     2341 tcctgcagga gaacctgaag gacatcatgc tgcagtgact cgaggcggcc gcaattcact
     2401 cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac
     2461 aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt
     2521 gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
     2581 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat
     2641 gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt
     2701 tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat
     2761 agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct
     2821 ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga
     2881 ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacct gcaccgtcga
     2941 gctagcatat ggatccatat aacttcgtat aatgtatgct atacgaagtt attacgatac
     3001 cgtcaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3061 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3121 atcttaggta cccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
     3181 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     3241 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     3301 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     3361 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     3421 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3481 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3541 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3601 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3661 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3721 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3781 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3841 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3901 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3961 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4021 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4081 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4141 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4201 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4261 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4321 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4381 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4441 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4501 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4561 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4621 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4681 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4741 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4801 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4861 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4921 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4981 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5041 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5101 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5161 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5221 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     5281 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc caagcttaat
     5341 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     5401 ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     5461 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     5521 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     5581 gcaggccctg ccatagccac tggccccgtg ggttagggac ggggtccccc attcgagaat
     5641 aaataacttc gtataaagta tcctatacga agttatcacc atgaccgagt acaagcccac
     5701 ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt
     5761 cgccgactac cccgccacgc gccacaccgt cgacccggac cgccacatcg agcgggtcac
     5821 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     5881 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     5941 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     6001 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     6061 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     6121 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     6181 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     6241 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga cgcccgcccc acgacccgca
     6301 gcgcccgacc gaaaggagcg cacgacccca tggctccgac cgaagccacc cggggcggcc
     6361 ccgccgaccc cgcacccgcc cccgaggccc accgactcta gagctcgctg atcagcctcg
     6421 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     6481 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     6541 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     6601 tgggaagaca atagcactag ttattaatag taatcaatta cggggtcatt agttcatagc
     6661 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     6721 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     6781 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     6841 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     6901 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     6961 ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc
     7021 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg
     7081 atgggggcgg gggggg
//
