LOCUS       sequence-246993-        5710 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Eukaryotic expression of the chimeric G protein Gqi9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5710)
  AUTHORS   Masharina A, Reymond L, Maurel D, Umezawa K, Johnsson K
  TITLE     A fluorescent sensor for GABA and synthetic GABA(B) receptor 
            ligands.
  JOURNAL   J Am Chem Soc. 2012 Nov 21;134(46):19026-34. doi: 10.1021/ja306320s.
            Epub 2012 Nov 5.
  PUBMED    23095089
REFERENCE   2  (bases 1 to 5710)
  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"; doi: "10.1021/ja306320s";
            journalName: "J Am Chem Soc"; date: "2012-11-21- 21"; volume: "134";
            issue: "46"; pages: "19026-34"
COMMENT     Sequence Label: pRK5-Gqi9
FEATURES             Location/Qualifiers
     source          1..5710
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     828..845
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     859..878
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2010..2019
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2023..2157
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2060..2079)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2114..2133
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2226..2583
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2226..2417
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2249..2269)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2434..2569
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2603..2620)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2603..2619)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2612..2634)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2832..3287
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2919..2938)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3129..3150
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3304..3323)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3423..3445
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3483..3501)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3569..3673
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3674..3742,3743..4534)
                     /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(3892..3911)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4705..5293
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5194..5213
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5447..5464
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5581..5602
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5617..5622,5623..5640,5641..5647)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5655..5671
                     /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     5660..5682
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5679..5695
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5679..5695
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      901 acctcggttc tatcgattga attccccggg gatccatggg gtgcaccctg agcgaggagg
      961 ccaaggaagc ccggaggatc aacgacgaga tcgagcggca gctgcgcagg gacaagcgcg
     1021 acgcccgccg ggagctcaag ctgctgctgc tggggacagg ggagagtggc aagtcgacct
     1081 tcatcaagca gatgaggatc atccacgggt cggactactc tgacgaagac aagcgcggct
     1141 tcaccaagct ggtgtatcag aacatcttca cggccatgca ggccatgatc agagcgatgg
     1201 acacactcaa gatcccatac aagtatgaac acaataaggc tcatgcacaa ttggttcgag
     1261 aggttgatgt ggagaaggtg tctgcttttg acgtccccga ctacgcggca ataaagagct
     1321 tgtggaatga tcctggaatc caggagtgct acgacagacg acgggaatat cagttatctg
     1381 actctaccaa atactatctg aatgacttgg accgtgtagc cgacccttcc tatctgccta
     1441 cacaacaaga cgtgcttaga gttcgagtcc ccactacagg gatcatcgaa tacccctttg
     1501 acttacaaag tgtcattttc agaatggtcg atgtaggggg ccaaaggtca gagagaagaa
     1561 aatggataca ctgctttgaa aatgtcacct ccatcatgtt tctagtagcg cttagcgaat
     1621 atgatcaagt tcttgtggag tcagacaatg agaaccgcat ggaggagagc aaagcactct
     1681 ttagaacaat tatcacctac ccctggttcc agaactcctc tgtgattctg ttcttaaaca
     1741 agaaagatct tctagaggag aaaatcatgt attcccacct agtcgactac ttcccagaat
     1801 atgatggacc ccagagagat gcccaggcag ctcgagaatt catcctgaaa atgttcgtgg
     1861 acctgaaccc cgacagtgac aaaatcatct actcccactt cacgtgcgcc acagataccg
     1921 agaacatccg cttcgtcttt gcagccgtca aggacaccat cctgcagctg aacctgaagt
     1981 actgtggcct ttgctaatgc ataagcttgg ccgccatggc ccaacttgtt tattgcagct
     2041 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     2101 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcggg
     2161 aattaattcg gcgcagcacc atggcctgaa ataacctctg aaagaggaac ttggttaggt
     2221 accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     2281 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     2341 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     2401 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     2461 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     2521 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     2581 aaaaagctgt taacagcttg gcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     2641 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     2701 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
     2761 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     2821 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2881 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     2941 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3001 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     3061 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3121 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc
     3181 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     3241 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     3301 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     3361 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     3421 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     3481 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     3541 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3601 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3661 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3721 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3781 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     3841 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     3901 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     3961 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     4021 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4081 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4141 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4201 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4261 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4321 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4381 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4441 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4501 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4561 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4621 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4681 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4741 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4801 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4861 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4921 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4981 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5041 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5101 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5161 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5221 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5281 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5341 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     5401 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     5461 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     5521 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     5581 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     5641 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgacatgat
     5701 tacgaattaa
//
