LOCUS       sequence-210330-        7060 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  GsX chimera with G-alpha-t C-terminus (Pertussis toxin insensitive 
            mutant).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7060)
  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 7060)
  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 Gst (Ser)
FEATURES             Location/Qualifiers
     source          1..7060
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     enhancer        60..363
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    508..619
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     608..625
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(761..1360)
                     /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(844..864)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1341..1360
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     protein_bind    complement(1365..1398)
                     /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)."
     polyA_signal    1610..1658
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        1731..1835
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1836..1904,1905..2696)
                     /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     complement(2054..2073)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2867..3455
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3356..3375
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3609..3626
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3743..3764
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3779..3784,3785..3802,3803..3809)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3817..3833
                     /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)."
     primer_bind     3822..3844
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3841..3857
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3841..3857
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3876..3896
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3878..3896
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    3915..4036
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3926..3945)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3980..3999
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(4049..4082)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     4428..4447
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    4446..4501
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     4547..4566
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     regulatory      5844..5853
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(5896..5915)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          5923..6939
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(5977..5999)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
ORIGIN
        1 gggggtgggg agggggggga gatggggaga gtgaagcaga acgtggggct cacctcgacc
       61 atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc
      121 atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta
      181 cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc
      241 cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac
      301 gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta
      361 acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa
      421 ctagtgctat tgtcttccca atcctccccc ttgctgtcct gccccacccc accccccaga
      481 atagaatgac acctactcag acaatgcgat gcaatttcct cattttatta ggaaaggaca
      541 gtgggagtgg caccttccag ggtcaaggaa ggcacggggg aggggcaaac aacagatggc
      601 tggcaactag aaggcacagt cgaggctgat cagcgagctc tagagtcggt gggcctcggg
      661 ggcgggtgcg gggtcggcgg ggccgccccg ggtggcttcg gtcggagcca tggggtcgtg
      721 cgctcctttc ggtcgggcgc tgcgggtcgt ggggcgggcg tcaggcaccg ggcttgcggg
      781 tcatgcacca ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga
      841 gccgctcgta gaaggggagg ttgcggggcg cggaggtctc caggaaggcg ggcaccccgg
      901 cgcgctcggc cgcctccact ccggggagca cgacggcgct gcccagaccc ttgccctggt
      961 ggtcgggcga gacgccgacg gtggccagga accacgcggg ctccttgggc cggtgcggcg
     1021 ccaggaggcc ttccatctgt tgctgcgcgg ccagccggga accgctcaac tcggccatgc
     1081 gcgggccgat ctcggcgaac accgcccccg cttcgacgct ctccggcgtg gtccagaccg
     1141 ccaccgcggc gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga
     1201 ggaagagttc ttgcagctcg gtgacccgct cgatgtggcg gtccgggtcg acggtgtggc
     1261 gcgtggcggg gtagtcggcg aacgcggcgg cgagggtgcg tacggcccgg gggacgtcgt
     1321 cgcgggtggc gaggcgcacc gtgggcttgt actcggtcat ggtgataact tcgtatagga
     1381 tactttatac gaagttattt attctcgaat gggggacccc gtccctaacc cacggggcca
     1441 gtggctatgg cagggcctgc cgccccgacg ttggctgcga gccctgggcc ttcacccgaa
     1501 cttggggggt ggggtgggga aaaggaagaa acgcgggcgt attggcccca atggggtctc
     1561 ggtggggtat cgacagagtg ccagccctgg gaccgaaccc cgcgtttatg aacaaacgac
     1621 ccaacacccg tgcgttttat tctgtctttt tattgccgtc atagcgcggg ttccttccgg
     1681 tattgtctcc ttccgtgttt attaagcttg gcacttttcg gggaaatgtg cgcggaaccc
     1741 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     1801 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     1861 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     1921 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     1981 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     2041 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     2101 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     2161 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     2221 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     2281 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     2341 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     2401 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     2461 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     2521 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     2581 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     2641 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     2701 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     2761 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     2821 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     2881 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     2941 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3001 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3061 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3121 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3181 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3241 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3301 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3361 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3421 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3481 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     3541 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     3601 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     3661 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     3721 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     3781 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     3841 caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac
     3901 aaaagctggg tacctaagat acattgatga gtttggacaa accacaacta gaatgcagtg
     3961 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     4021 ctgcaataaa caagttgacg gtatcgtaat aacttcgtat agcatacatt atacgaagtt
     4081 atatggatcc atatgctagc tcgacggtgc aggtcgaggg atctccataa gagaagaggg
     4141 acagctatga ctgggagtag tcaggagagg aggaaaaatc tggctagtaa aacatgtaag
     4201 gaaaatttta gggatgttaa agaaaaaaat aacacaaaac aaaatataaa aaaaatctaa
     4261 cctcaagtca aggcttttct atggaataag gaatggacag cagggggctg tttcatatac
     4321 tgatgacctc tttatagcca acctttgttc atggcagcca gcatatgggc atatgttgcc
     4381 aaactctaaa ccaaatactc attctgatgt tttaaatgat ttgccctccc atatgtcctt
     4441 ccgagtgaga gacacaaaaa attccaacac actattgcaa tgaaaataaa tttcctttat
     4501 tagccagaag tcagatgctc aaggggcttc atgatgtccc cataattttt ggcagaggga
     4561 aaaagatctc agtggtattt gtgagccagg gcattggcca caccagccac caccttctga
     4621 taggcagcct gcacctgagg agtgaattgc ggccgcctcg agtcagaaga ggccagagtc
     4681 tttgaggttc tccttgatga tgatgtcacg gcagtcgttg aacacacggc ggatgttctc
     4741 agtgtccaca gcgcaggtga aatgagggta gcagtagtga cgcccatctc cactggcagt
     4801 gctgatcctc agaaactcat ctcgaatgaa gtacttggcc cgggtcacgc gtgggtcctc
     4861 tccgggctcg ggagtagcat cctcaggagt agtgtagcga gcaaattctg gaaagtagtc
     4921 ctcaatcttc gatttcccag caaggacttt ctcagcgagc agatcttgct tgttgaggaa
     4981 caggatcaca gagatggtgc gcagccatct gttgttccag atgctcttga agaggttcag
     5041 agcctcctgc aggcggttgg tctggttgtc ctcccggatg accatgttgt agctgctgct
     5101 ggccaccacg aagatgatgg cagtcacatc gttgaagcac tggatccact tgcggcgttc
     5161 atcgcgctgg ccacccacgt caaacatgtg gaagttgact ttgtccacct ggaacttggt
     5221 ctcaaagatt ccagaagtca ggacacggca gcgaagcagg tcctgatcgc tcggcacata
     5281 gtcagcctgc ttgatcacgt cgatcttgtc caggaagtac tgggcacagt caatcagctg
     5341 gtactcgttg gagcgttcgt agcaggcacg cactccttca tcctcccaca gagccttggc
     5401 atgctcatag aattcgggag ggaagtcaaa gtcaggcacg ttcatcacac tcagaatgta
     5461 gtccactctg aactggttct cggggttggc cagctccacg gggggcacca ggttgctcat
     5521 ggcggccaca atggtttcaa tcgcctcttt caggttgttt ttgatgtcct gcactttggt
     5581 tgccttctca ccatcgctgt tgctccttgc agcctgcggg tcctcttcgc cgccctctcc
     5641 attaaaccca ttaacatgca ggatcctcat ctgcttcaca atggtgcttt taccagattc
     5701 tccagcaccc agcagcagca ggcggtgcgt ggcccggtag acctgcttgt ccttctgcag
     5761 ctgcttctcg atctttttgt tggcctcacg ctgcgccttc tcctcgttgc gctggtcctc
     5821 ggtcttactg ttcccgaggc agcccatggt ggcgcgccgg tggtcgagca attctttgcc
     5881 aaaatgatga gacagcacaa taaccagcac gttgcccagg agctgtagga aaaagaagaa
     5941 ggcatgaaca tggttagcag aggctctaga gccgccggtc acacgccaga agccgaaccc
     6001 cgccctgccc cgtccccccc gaaggcagcc gtccccccgc ggacagcccc gaggctggag
     6061 agggagaagg ggacggcggc gcggcgacgc acgaaggccc tccccgccca tttccttcct
     6121 gccggcgccg caccgcttcg ccccgcgccc gctagagggg gtgcggcggc gcctcccaga
     6181 tttcggctcc gcacagattt gggacaaagg aagtccctgc gccctctcgc acgattacca
     6241 taaaaggcaa tggctgcggc tcgccgcgcc tcgacagccg ccggcgctcc gggggccgcc
     6301 gcgcccctcc cccgagccct ccccggcccg aggcggcccc gccccgcccg gcacccccac
     6361 ctgccgccac cccccgcccg gcacggcgag ccccgcgcca cgccccgtac ggagccccgc
     6421 acccgaagcc gggccgtgct cagcaactcg gggagggggg tgcagggggg gttgcagccc
     6481 gaccgacgcg cccacacccc ctgctcaccc ccccacgcac acaccccgca cgcagccttt
     6541 gttcccctcg cagccccccc cgcaccgcgg ggcaccgccc ccggccgcgc tcccctcgcg
     6601 cacactgcgg agcgcacaaa gccccgcgcc gcgcccgcag cgctcacagc cgccgggcag
     6661 cgcggagccg cacgcggcgc tccccacgca cacacacacg cacgcacccc ccgagccgct
     6721 ccccccgcac aaagggccct cccggagccc ctcaaggctt tcacgcagcc acagaaaaga
     6781 aacaagccgt cattaaacca agcgctaatt acagcccgga ggagaagggc cgtcccgccc
     6841 gctcacctgt gggagtaacg cggtcagtca gagccggggc gggcggcgcg aggcggcggc
     6901 ggagcggggc acggggcgaa ggcagcgcgc agcgactccc gcccgccgcg cgcttcgctt
     6961 tttatagggc cgccgccgcc gccgcctcgc cataaaagga aactttcgga gcgcgccgct
     7021 ctgattggct gccgccgcac ctctccgcct cgccccgccc
//
