LOCUS       sequence-210400-        7102 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  GsX chimera with G-alpha-15 C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7102)
  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 7102)
  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 Gs15
FEATURES             Location/Qualifiers
     source          1..7102
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          111..1127
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1051..1073
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1135..1154
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1197..1206
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2484..2503)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2549..2604
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2603..2622)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     protein_bind    2968..3001
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    3014..3135
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3051..3070)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3105..3124
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3154..3174)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3154..3172)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3193..3209)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3193..3209)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3206..3228)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3217..3233
                     /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(3241..3247,3248..3265,3266..3271))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3286..3307
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3424..3441)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3595..4183)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3675..3694)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4354..5145,5146..5214))
                     /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     4977..4996
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5215..5319)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    5392..5440
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    5652..5685
                     /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             5690..6289
                     /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(5690..5709)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6186..6206
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(6425..6442)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6431..6542
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        6687..6990
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg
       61 cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg
      121 cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct
      181 gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta
      241 attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa agccttgagg
      301 ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt
      361 gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg
      421 gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc
      481 cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg
      541 ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc
      601 gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc
      661 tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg gggccgcctc
      721 gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag
      781 gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc
      841 ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg
      901 ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg
      961 cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg
     1021 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
     1081 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg
     1141 tgctggttat tgtgctgtct catcattttg gcaaagaatt gctcgaccac cggcgcgcca
     1201 ccatgggctg cctcgggaac agtaagaccg aggaccagcg caacgaggag aaggcgcagc
     1261 gtgaggccaa caaaaagatc gagaagcagc tgcagaagga caagcaggtc taccgggcca
     1321 cgcaccgcct gctgctgctg ggtgctggag aatctggtaa aagcaccatt gtgaagcaga
     1381 tgaggatcct gcatgttaat gggtttaatg gagagggcgg cgaagaggac ccgcaggctg
     1441 caaggagcaa cagcgatggt gagaaggcaa ccaaagtgca ggacatcaaa aacaacctga
     1501 aagaggcgat tgaaaccatt gtggccgcca tgagcaacct ggtgcccccc gtggagctgg
     1561 ccaaccccga gaaccagttc agagtggact acattctgag tgtgatgaac gtgcctgact
     1621 ttgacttccc tcccgaattc tatgagcatg ccaaggctct gtgggaggat gaaggagtgc
     1681 gtgcctgcta cgaacgctcc aacgagtacc agctgattga ctgtgcccag tacttcctgg
     1741 acaagatcga cgtgatcaag caggctgact atgtgccgag cgatcaggac ctgcttcgct
     1801 gccgtgtcct gacttctgga atctttgaga ccaagttcca ggtggacaaa gtcaacttcc
     1861 acatgtttga cgtgggtggc cagcgcgatg aacgccgcaa gtggatccag tgcttcaacg
     1921 atgtgactgc catcatcttc gtggtggcca gcagcagcta caacatggtc atccgggagg
     1981 acaaccagac caaccgcctg caggaggctc tgaacctctt caagagcatc tggaacaaca
     2041 gatggctgcg caccatctct gtgatcctgt tcctcaacaa gcaagatctg ctcgctgaga
     2101 aagtccttgc tgggaaatcg aagattgagg actactttcc agaatttgct cgctacacta
     2161 ctcctgagga tgctactccc gagcccggag aggacccacg cgtgacccgg gccaagtact
     2221 tcattcgaga tgagtttctg aggatcagca ctgccagtgg agatgggcgt cactactgct
     2281 accctcattt cacctgcgct gtggacactg agaacatccg ccgtgtgttc aacgactgcc
     2341 gtgactcggt gctcgcccgc tacctggacg agatcaacct gctgtgactc gaggcggccg
     2401 caattcactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc
     2461 ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg ggacatcatg
     2521 aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg
     2581 tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa
     2641 catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct ggctgccatg
     2701 aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc
     2761 ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta
     2821 tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc
     2881 ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc cctcgacctg
     2941 caccgtcgag ctagcatatg gatccatata acttcgtata atgtatgcta tacgaagtta
     3001 ttacgatacc gtcaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     3061 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     3121 catcaatgta tcttaggtac ccagcttttg ttccctttag tgagggttaa ttgcgcgctt
     3181 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     3241 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     3301 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     3361 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     3421 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3481 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3541 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3601 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3661 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3721 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3781 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3841 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3901 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3961 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4021 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4081 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4141 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4201 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4261 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4321 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4381 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4441 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4501 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     4561 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4621 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     4681 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4741 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4801 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4861 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4921 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4981 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5041 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5101 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5161 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5221 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     5281 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     5341 aagcttaata aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     5401 aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     5461 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     5521 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     5581 gtcggggcgg caggccctgc catagccact ggccccgtgg gttagggacg gggtccccca
     5641 ttcgagaata aataacttcg tataaagtat cctatacgaa gttatcacca tgaccgagta
     5701 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc
     5761 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc gccacatcga
     5821 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     5881 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     5941 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     6001 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     6061 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     6121 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
     6181 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     6241 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca
     6301 cgacccgcag cgcccgaccg aaaggagcgc acgaccccat ggctccgacc gaagccaccc
     6361 ggggcggccc cgccgacccc gcacccgccc ccgaggccca ccgactctag agctcgctga
     6421 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     6481 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     6541 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     6601 ggggaggatt gggaagacaa tagcactagt tattaatagt aatcaattac ggggtcatta
     6661 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     6721 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     6781 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     6841 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     6901 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     6961 atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact
     7021 ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt
     7081 tgtgcagcga tgggggcggg gg
//
