LOCUS       sequence-206188-        7184 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GsX chimera with G-alpha-i C-terminus (Pertussis toxin insensitive 
            mutant).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7184)
  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 7184)
  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 (Ser)
FEATURES             Location/Qualifiers
     source          1..7184
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    52..100
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    312..345
                     /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             350..949
                     /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(350..369)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     846..866
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(1085..1102)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1091..1202
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        1347..1650
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1652..1929
                     /label=chicken beta-actin promoter
     intron          1930..2946
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2870..2892
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2954..2973
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      3016..3025
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4329..4348)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4394..4449
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4448..4467)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     protein_bind    4812..4845
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    4858..4979
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4895..4914)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4949..4968
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4998..5018)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4998..5016)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5037..5053)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5037..5053)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5050..5072)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5061..5077
                     /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(5085..5091,5092..5109,5110..5115))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5130..5151
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5268..5285)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5439..6027)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5519..5538)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6198..6989,6990..7058))
                     /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     6821..6840
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7059..7163)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aagcttaata aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
       61 aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
      121 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
      181 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
      241 gtcggggcgg caggccctgc catagccact ggccccgtgg gttagggacg gggtccccca
      301 ttcgagaata aataacttcg tataaagtat cctatacgaa gttatcacca tgaccgagta
      361 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc
      421 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc gccacatcga
      481 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
      541 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
      601 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
      661 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
      721 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
      781 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
      841 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
      901 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca
      961 cgacccgcag cgcccgaccg aaaggagcgc acgaccccat ggctccgacc gaagccgacc
     1021 cgggcggccc cgccgacccc gcacccgccc ccgaggccca ccgactctag agctcgctga
     1081 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     1141 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     1201 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     1261 ggggaggatt gggaagacaa tagcactagt tattaatagt aatcaattac ggggtcatta
     1321 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     1381 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     1441 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     1501 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     1561 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1621 atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact
     1681 ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt
     1741 tgtgcagcga tgggggcggg gggggggggg gggcgcgcgc caggcggggc ggggcggggc
     1801 gaggggcggg gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc
     1861 cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg
     1921 cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg
     1981 cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc
     2041 ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg
     2101 ctgcgtgaaa gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg
     2161 ggtgcgtgcg tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc
     2221 tgtgagcgct gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc
     2281 gcggccgggg gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg
     2341 cggggtgtgt gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc
     2401 cccctgcacc cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt
     2461 acggggcgtg gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg
     2521 ggcggggcgg ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggcccccgg
     2581 agcgccggcg gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc
     2641 gagagggcgc agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc
     2701 cgccgcaccc cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg
     2761 ggcggggagg gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg
     2821 ggctgtccgc ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc
     2881 tggcgtgtga ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc
     2941 ctacagctcc tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattg
     3001 ctcgaccacc ggcgcgccac catgggctgc ctcgggaaca gtaagaccga ggaccagcgc
     3061 aacgaggaga aggcgcagcg tgaggccaac aaaaagatcg agaagcagct gcagaaggac
     3121 aagcaggtct accgggccac gcaccgcctg ctgctgctgg gtgctggaga atctggtaaa
     3181 agcaccattg tgaagcagat gaggatcctg catgttaatg ggtttaatgg agagggcggc
     3241 gaagaggacc cgcaggctgc aaggagcaac agcgatggtg agaaggcaac caaagtgcag
     3301 gacatcaaaa acaacctgaa agaggcgatt gaaaccattg tggccgccat gagcaacctg
     3361 gtgccccccg tggagctggc caaccccgag aaccagttca gagtggacta cattctgagt
     3421 gtgatgaacg tgcctgactt tgacttccct cccgaattct atgagcatgc caaggctctg
     3481 tgggaggatg aaggagtgcg tgcctgctac gaacgctcca acgagtacca gctgattgac
     3541 tgtgcccagt acttcctgga caagatcgac gtgatcaagc aggctgacta tgtgccgagc
     3601 gatcaggacc tgcttcgctg ccgtgtcctg acttctggaa tctttgagac caagttccag
     3661 gtggacaaag tcaacttcca catgtttgac gtgggtggcc agcgcgatga acgccgcaag
     3721 tggatccagt gcttcaacga tgtgactgcc atcatcttcg tggtggccag cagcagctac
     3781 aacatggtca tccgggagga caaccagacc aaccgcctgc aggaggctct gaacctcttc
     3841 aagagcatct ggaacaacag atggctgcgc accatctctg tgatcctgtt cctcaacaag
     3901 caagatctgc tcgctgagaa agtccttgct gggaaatcga agattgagga ctactttcca
     3961 gaatttgctc gctacactac tcctgaggat gctactcccg agcccggaga ggacccacgc
     4021 gtgacccggg ccaagtactt cattcgagat gagtttctga ggatcagcac tgccagtgga
     4081 gatgggcgtc actactgcta ccctcatttc acctgcgctg tggacactga gaacatccgc
     4141 cgtgtgttca acgactgccg tgatgtcatc ataaaaaata atctaaaaga ttctggtctc
     4201 ttttgaaccg gtctgcagcc taggcttaag cgctcgaggc ggccgcaatt cactcctcag
     4261 gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac
     4321 cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat
     4381 ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg
     4441 tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat
     4501 ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta
     4561 taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa
     4621 gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca
     4681 tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc
     4741 ccagtcatag ctgtccctct tctcttatga agatccctcg acctgagcgt cgagctagca
     4801 tatggatcca tataacttcg tataatgtat gctatacgaa gttattacga taccgtcaac
     4861 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     4921 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttag
     4981 gtacccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
     5041 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     5101 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5161 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5221 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5281 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5341 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5401 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5461 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5521 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5581 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5641 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5701 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5761 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5821 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     5881 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5941 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6001 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6061 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6121 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6181 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6241 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6301 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6361 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6421 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6481 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6541 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6601 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6661 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6721 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6781 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6841 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6901 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6961 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7021 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7081 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7141 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgcc
//
