LOCUS       sequence-210315-        7110 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  GsX chimera with G-alpha-z C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7110)
  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 7110)
  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 Gsz
FEATURES             Location/Qualifiers
     source          1..7110
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          112..1128
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1052..1074
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1136..1155
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1198..1207
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2485..2504)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2550..2605
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2604..2623)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     protein_bind    2969..3002
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    3015..3136
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3052..3071)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3106..3125
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3155..3175)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3155..3173)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3194..3210)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3194..3210)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3207..3229)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3218..3234
                     /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(3242..3248,3249..3266,3267..3272))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3287..3308
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3425..3442)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3596..4184)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3676..3695)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4355..5146,5147..5215))
                     /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     4978..4997
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5216..5320)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    5393..5441
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    5653..5686
                     /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             5691..6290
                     /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(5691..5710)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6187..6207
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(6426..6443)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6432..6543
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        6688..6991
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg
       61 gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct
      121 gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc
      181 tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt
      241 aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga aagccttgag
      301 gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg
      361 tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc
      421 ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc
      481 ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg
      541 gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca cccccctccc
      601 cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg
      661 ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct
      721 cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga
      781 ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc gcagggactt
      841 cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc
      901 gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt
      961 gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac
     1021 ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc
     1081 tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac
     1141 gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tgctcgacca ccggcgcgcc
     1201 accatgggct gcctcgggaa cagtaagacc gaggaccagc gcaacgagga gaaggcgcag
     1261 cgtgaggcca acaaaaagat cgagaagcag ctgcagaagg acaagcaggt ctaccgggcc
     1321 acgcaccgcc tgctgctgct gggtgctgga gaatctggta aaagcaccat tgtgaagcag
     1381 atgaggatcc tgcatgttaa tgggtttaat ggagagggcg gcgaagagga cccgcaggct
     1441 gcaaggagca acagcgatgg tgagaaggca accaaagtgc aggacatcaa aaacaacctg
     1501 aaagaggcga ttgaaaccat tgtggccgcc atgagcaacc tggtgccccc cgtggagctg
     1561 gccaaccccg agaaccagtt cagagtggac tacattctga gtgtgatgaa cgtgcctgac
     1621 tttgacttcc ctcccgaatt ctatgagcat gccaaggctc tgtgggagga tgaaggagtg
     1681 cgtgcctgct acgaacgctc caacgagtac cagctgattg actgtgccca gtacttcctg
     1741 gacaagatcg acgtgatcaa gcaggctgac tatgtgccga gcgatcagga cctgcttcgc
     1801 tgccgtgtcc tgacttctgg aatctttgag accaagttcc aggtggacaa agtcaacttc
     1861 cacatgtttg acgtgggtgg ccagcgcgat gaacgccgca agtggatcca gtgcttcaac
     1921 gatgtgactg ccatcatctt cgtggtggcc agcagcagct acaacatggt catccgggag
     1981 gacaaccaga ccaaccgcct gcaggaggct ctgaacctct tcaagagcat ctggaacaac
     2041 agatggctgc gcaccatctc tgtgatcctg ttcctcaaca agcaagatct gctcgctgag
     2101 aaagtccttg ctgggaaatc gaagattgag gactactttc cagaatttgc tcgctacact
     2161 actcctgagg atgctactcc cgagcccgga gaggacccac gcgtgacccg ggccaagtac
     2221 ttcattcgag atgagtttct gaggatcagc actgccagtg gagatgggcg tcactactgc
     2281 taccctcatt tcacctgcgc tgtggacact gagaacatcc gccgtgtgtt caacgactgc
     2341 cgtgacgtca tcatacagaa caatctcaag tacattggcc tttgctgact cgaggcggcc
     2401 gcaattcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc
     2461 cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat
     2521 gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt
     2581 gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa
     2641 acatcagaat gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat
     2701 gaacaaaggt tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc
     2761 cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt
     2821 atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct
     2881 cctctcctga ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacct
     2941 gcaccgtcga gctagcatat ggatccatat aacttcgtat aatgtatgct atacgaagtt
     3001 attacgatac cgtcaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3061 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3121 tcatcaatgt atcttaggta cccagctttt gttcccttta gtgagggtta attgcgcgct
     3181 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3241 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3301 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3361 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3421 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3481 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3541 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3601 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3661 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3721 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3781 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3841 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3901 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3961 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4021 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4081 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4141 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4201 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4261 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4321 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4381 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4441 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4501 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4561 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4621 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4681 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4741 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4801 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4861 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4921 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4981 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5041 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5101 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5161 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5221 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5281 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5341 caagcttaat aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     5401 aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     5461 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     5521 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     5581 cgtcggggcg gcaggccctg ccatagccac tggccccgtg ggttagggac ggggtccccc
     5641 attcgagaat aaataacttc gtataaagta tcctatacga agttatcacc atgaccgagt
     5701 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta cgcaccctcg
     5761 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgacccggac cgccacatcg
     5821 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     5881 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     5941 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     6001 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     6061 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     6121 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     6181 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     6241 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga cgcccgcccc
     6301 acgacccgca gcgcccgacc gaaaggagcg cacgacccca tggctccgac cgaagccacc
     6361 cggggcggcc ccgccgaccc cgcacccgcc cccgaggccc accgactcta gagctcgctg
     6421 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     6481 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     6541 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     6601 gggggaggat tgggaagaca atagcactag ttattaatag taatcaatta cggggtcatt
     6661 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     6721 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     6781 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     6841 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     6901 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     6961 catctacgta ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac
     7021 tctccccatc tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt
     7081 ttgtgcagcg atgggggcgg gggggggggg
//
