LOCUS       sequence-210404-        7057 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  GsX chimera with G-alpha-13 C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7057)
  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 7057)
  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 Gs13
FEATURES             Location/Qualifiers
     source          1..7057
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          60..1076
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1000..1022
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1084..1103
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1146..1155
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2433..2452)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2498..2553
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2552..2571)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     protein_bind    2917..2950
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    2963..3084
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3000..3019)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3054..3073
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3103..3123)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3103..3121)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3142..3158)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3142..3158)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3155..3177)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3166..3182
                     /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(3190..3196,3197..3214,3215..3220))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3235..3256
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3373..3390)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3544..4132)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3624..3643)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4303..5094,5095..5163))
                     /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     4926..4945
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5164..5268)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    5341..5389
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    5601..5634
                     /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             5639..6238
                     /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(5639..5658)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6135..6155
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(6374..6391)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6380..6491
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        6636..6939
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
       61 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
      121 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      181 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
      241 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg
      301 tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct
      361 gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg
      421 gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
      481 gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc
      541 cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg
      601 gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg
      661 ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggcccccgg agcgccggcg
      721 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc
      781 agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc
      841 cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg
      901 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc
      961 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga
     1021 ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc
     1081 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattg ctcgaccacc
     1141 ggcgcgccac catgggctgc ctcgggaaca gtaagaccga ggaccagcgc aacgaggaga
     1201 aggcgcagcg tgaggccaac aaaaagatcg agaagcagct gcagaaggac aagcaggtct
     1261 accgggccac gcaccgcctg ctgctgctgg gtgctggaga atctggtaaa agcaccattg
     1321 tgaagcagat gaggatcctg catgttaatg ggtttaatgg agagggcggc gaagaggacc
     1381 cgcaggctgc aaggagcaac agcgatggtg agaaggcaac caaagtgcag gacatcaaaa
     1441 acaacctgaa agaggcgatt gaaaccattg tggccgccat gagcaacctg gtgccccccg
     1501 tggagctggc caaccccgag aaccagttca gagtggacta cattctgagt gtgatgaacg
     1561 tgcctgactt tgacttccct cccgaattct atgagcatgc caaggctctg tgggaggatg
     1621 aaggagtgcg tgcctgctac gaacgctcca acgagtacca gctgattgac tgtgcccagt
     1681 acttcctgga caagatcgac gtgatcaagc aggctgacta tgtgccgagc gatcaggacc
     1741 tgcttcgctg ccgtgtcctg acttctggaa tctttgagac caagttccag gtggacaaag
     1801 tcaacttcca catgtttgac gtgggtggcc agcgcgatga acgccgcaag tggatccagt
     1861 gcttcaacga tgtgactgcc atcatcttcg tggtggccag cagcagctac aacatggtca
     1921 tccgggagga caaccagacc aaccgcctgc aggaggctct gaacctcttc aagagcatct
     1981 ggaacaacag atggctgcgc accatctctg tgatcctgtt cctcaacaag caagatctgc
     2041 tcgctgagaa agtccttgct gggaaatcga agattgagga ctactttcca gaatttgctc
     2101 gctacactac tcctgaggat gctactcccg agcccggaga ggacccacgc gtgacccggg
     2161 ccaagtactt cattcgagat gagtttctga ggatcagcac tgccagtgga gatgggcgtc
     2221 actactgcta ccctcatttc acctgcgctg tggacactga gaacatccgc cgtgtgttca
     2281 acgactgccg tgacactatt ctgcatgaca acctcaagca gcttatgcta cagtgactcg
     2341 aggcggccgc aattcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg
     2401 gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg
     2461 gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt
     2521 gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat
     2581 catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg
     2641 gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct
     2701 gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt
     2761 tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga
     2821 tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc
     2881 ctcgacctgc accgtcgagc tagcatatgg atccatataa cttcgtataa tgtatgctat
     2941 acgaagttat tacgataccg tcaacttgtt tattgcagct tataatggtt acaaataaag
     3001 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3061 gtccaaactc atcaatgtat cttaggtacc cagcttttgt tccctttagt gagggttaat
     3121 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     3181 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     3241 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     3301 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3361 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3421 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3481 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3541 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3601 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3661 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3721 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3781 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3841 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3901 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3961 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4021 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4081 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4141 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4201 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4261 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4321 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4381 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4441 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4501 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4561 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4621 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4681 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4741 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4801 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4861 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4921 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4981 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5041 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5101 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5161 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5221 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5281 aaaagtgcca agcttaataa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     5341 cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg
     5401 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     5461 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     5521 gcagccaacg tcggggcggc aggccctgcc atagccactg gccccgtggg ttagggacgg
     5581 ggtcccccat tcgagaataa ataacttcgt ataaagtatc ctatacgaag ttatcaccat
     5641 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg
     5701 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg acccggaccg
     5761 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     5821 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     5881 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     5941 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     6001 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     6061 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     6121 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     6181 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg
     6241 cccgccccac gacccgcagc gcccgaccga aaggagcgca cgaccccatg gctccgaccg
     6301 aagccacccg gggcggcccc gccgaccccg cacccgcccc cgaggcccac cgactctaga
     6361 gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     6421 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     6481 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     6541 gacagcaagg gggaggattg ggaagacaat agcactagtt attaatagta atcaattacg
     6601 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     6661 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     6721 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     6781 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     6841 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     6901 tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc
     6961 tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt
     7021 taattatttt gtgcagcgat gggggcgggg ggggggg
//
