LOCUS       sequence-385866-        8203 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes a G alpha subunit (GNAS2) with RLuc8, a G gamma subunit 
            (GNG9) with GFP2 and a G beta subunit (GNB3) as optimal components 
            of a BRET2 biosensor for studying heterotrimeric G proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8203)
  AUTHORS   Olsen RHJ, DiBerto JF, English JG, Glaudin AM, Krumm BE, Slocum ST, 
            Che T, Gavin AC, McCorvy JD, Roth BL, Strachan RT
  TITLE     TRUPATH, an open-source biosensor platform for interrogating the 
            GPCR transducerome.
  JOURNAL   Nat Chem Biol. 2020 May 4. pii: 10.1038/s41589-020-0535-8. doi: 
            10.1038/s41589-020-0535-8.
  PUBMED    32367019
REFERENCE   2  (bases 1 to 8203)
  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: "Nat Chem 
            Biol. 2020 May 4. pii: 10.1038/s41589-020-0535-8. doi: 
            10.1038/s41589-020-0535-8."
COMMENT     Sequence Label: TRUPATH Triple (bidirectional) GasS
FEATURES             Location/Qualifiers
     source          1..8203
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(68..523)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(205..226)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     417..436
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     631..648
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    659..740
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(3031..3234)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3060..3080)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3235..3538
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             4705..4758
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     polyA_signal    5845..5926
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(6215..6803)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6295..6314)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6974..7765,7766..7834))
                     /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     7597..7616
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7835..7939)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8007..8025
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8063..8085)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     join(8185..8203,1)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
       61 catcaggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
      121 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
      181 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
      241 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
      301 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
      361 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
      421 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
      481 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtccattcg ccattcaggc
      541 tgcgcaactg ttgggaaggg cgatctggat aaccgtatta ccgcctttga gtgagctgat
      601 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agctcggggc
      661 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
      721 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
      781 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt
      841 atgatcctct ggagatatcg tcgacaagct tatcgatgcg gccgctcaca gcagctcata
      901 ctgtctcagg tgcatccgct ggatgatgtc gcgacagtca ttgaacactc tccggatgtt
      961 ctctgtgtcc acggcacagg taaagtgggg atagcagtag tgccttccgt cgccgcttgc
     1021 ggtagagatt ctcagaaact catcccggat gaagtacttg gctcttgtca cccgtgggtc
     1081 ctctcctggc tcaggggttg catcctcagg tgtggtatag cgggcaaact cggggaagta
     1141 gtcctcgatc ttgctcttgc cggccagcac cttctcggcc agcagatcct gcttattcag
     1201 gaacaggatc acgctgatgg tgcgcagcca cctattgttc cagatagact taaacaggtt
     1261 cagggcctcc tgcaggcgat ttgtctggtt gtcctccctg atgaccatat tgtaggagct
     1321 agaggccacc acaaagatga tggcggtcac atcgttgaaa cactgaatcc acttcctcct
     1381 ctcgtccctc tgtcctccca catcaaacat gtggaagttc accttgtcca cctgaaactt
     1441 tgtctcgaag atgccagagg tcagcactct gcaccgcagc agatcctggt cggaaggcac
     1501 atagtcggcc tgcttgatca catcgatctt gtccaggaag tactgggcac aatcgatcag
     1561 ctgatactca ttgctcctct cgtagcatgc cctcacgccc tcatcctccc acagggcctt
     1621 ggcgtgctca taaaactcgg gtggaaaatc gaagtcgggc acggagcctc ctcctccgga
     1681 ctgctcgttc ttcagcaccc tctccacgaa ggacttgatg tacttgccca tctcatcagg
     1741 ggcgtcctcc tgcagaaagt gcaggccctt caccttcaca aactcggtat tggggaactt
     1801 cttggcgccc tccacgatgg cgttgctaaa gaagcctggg tcagactcga tgaacagctt
     1861 aggcagatcg tcagaggctc tcagataggc attgtagttc cgcacgatct gcaccacatc
     1921 tggcttgccg cccttcacca gggggatctc tcttggccag gacagggtag gtctccgcac
     1981 ctcgcccttc tccttaaaag gctccaggta tgctgcgaac tcctctggct ccagcttcct
     2041 catgatcttg gatggcagca ctgtctccac aaagaaattg ttctccagca ccatcttctc
     2101 gccctcctcg ctcttgatca gggcgatgtc ctcctcgatg tcgggccact cgtcccagga
     2161 ctcgatcacg tccaccacgc tctccatgtg cacgatggcc ttgattctat cctggtgctc
     2221 ataggcgtag tggaatgcca gtgctgctcc ccagtcgtgt cccacaaaga tgatcttctt
     2281 tggcagattc agcagctcga accaggctgt cagatacttg tagtggtcca gcagccgata
     2341 gctgccgttg ccggacttgc cgctcttgcc catgccgatc agatcaggga tgatacatct
     2401 ggccacaggc tcgatgtggg gcaccacgtg cctccacagg taggagctgg tggcattgcc
     2461 gtgcaggaag atcacggcgt tctcggcgtg cttctcagag tcatagtaat tgataaagga
     2521 atccagcacg ttcatctgct tgcaccttgc ccaccactgt ggtcctgtga tcatgcgctt
     2581 cctctgctct gggtcataca ccttagatgc catggagcct cctcctccag aattcatcac
     2641 ggacaggatg taatccaccc tgaactgatt ctcggggttt gccagctcca cagggggcac
     2701 caggttgctc attgctgcca cgattgtctc gatggcctcc ttcagattgt tcttgatgtc
     2761 ctgcaccttg gtggccttct cagaatcgcc attaaagccg ttcacgtgca ggattctcat
     2821 ctgcttcacg attgtgctct tgccagactc tcctgctccc agcagcagca gcctgtgggt
     2881 ggccctgtac acctgcttgt ccttctgcag ctgcttctcg atcttcttgt tggcctcgcg
     2941 ctgggccttc tcctcattcc tctgatcctc tgtcttggag ttgcccaggc agcccatgga
     3001 tccctagcgg atctgacggt tcactaaacc agctctgctt atatagacct cccaccgtac
     3061 acgcctaccg cccatttgcg tcaatggggc ggagttgtta cgacattttg gaaagtcccg
     3121 ttgattttgg tgccaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc
     3181 ccgtgagtca aaccgctatc cacgcccatt gatgtactgc caaaaccgca tcaccatggt
     3241 aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta
     3301 ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg
     3361 catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg
     3421 acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa
     3481 tgggcggggg tcgttgggcg gtcagccagg cgagccattt accgtaagtt atgtaacgcg
     3541 gaactccata tatgggctat gaactaatga ccccgtaatt gattactatt aataactact
     3601 cgagtatgtc gaggtggcgt gtacggtggg aggcctatat aagcagagct cgtttagtga
     3661 accgtcagat cgcctggaga attcatggga gagatggagc agctgaggca ggaggcagag
     3721 cagctgaaga agcagatcgc cgacgccaga aaggcctgcg cagatgtgac cctggcagag
     3781 ctggtgagcg gactggaggt ggtgggaagg gtgcagatga ggaccaggag aacactgagg
     3841 ggccacctgg ccaagatcta cgccatgcac tgggccacag actctaagct gctggtgtcc
     3901 gcctctcagg acggcaagct gatcgtgtgg gatagctata ccacaaacaa ggtgcacgca
     3961 atccccctgc gcagctcctg ggtcatgacc tgtgcctacg ccccttccgg aaatttcgtg
     4021 gcatgcggag gactggataa catgtgctct atctataatc tgaagagccg ggagggcaac
     4081 gtgaaggtgt ctagagagct gagcgcccac accggctacc tgtcttgctg tcggttcctg
     4141 gacgataaca atatcgtgac atctagcggc gacaccacat gcgcactgtg ggacatcgag
     4201 accggacagc agaagacagt gtttgtgggc cacacaggcg actgtatgtc cctggccgtg
     4261 tctccagatt tcaatctgtt tatctccgga gcatgcgacg catctgccaa gctgtgggat
     4321 gtgcgggagg gcacctgtag acagaccttc acaggccacg agagcgacat caacgccatc
     4381 tgcttctttc ccaatggcga ggccatctgc acaggcagcg acgatgcctc ctgtaggctg
     4441 ttcgacctgc gcgccgatca ggagctgatc tgcttttctc acgagagcat catctgtggc
     4501 atcacctccg tggcctttag cctgtccggc aggctgctgt tcgcaggata tgacgacttc
     4561 aactgcaacg tgtgggactc catgaagtct gagagggtgg gcatcctgag cggccacgat
     4621 aatcgcgtgt cctgtctggg agtgaccgca gacggaatgg cagtggccac aggcagctgg
     4681 gattcctttc tgaagatctg gaacgagggc agaggaagtc ttctaacatg cggtgacgtg
     4741 gaggagaatc ccggccctac cggtatggtg tccaagggag aggagctgtt caccggagtg
     4801 gtgcctatcc tggtggagct ggacggcgat gtgaatggcc acaagtttag cgtgtccgga
     4861 gagggagagg gcgacgcaac ctacggcaag ctgacactga agttcatctg caccacaggc
     4921 aagctgcccg tgccttggcc aaccctggtg accacactgt cctacggcgt gcagtgtttt
     4981 tctaggtatc ctgaccacat gaagcagcac gatttcttta agtctgccat gccagagggc
     5041 tacgtgcagg agcggacaat cttctttaag gacgatggca actataagac cagagccgag
     5101 gtgaagttcg agggcgacac actggtgaac aggatcgagc tgaagggcat cgactttaag
     5161 gaggatggca atatcctggg ccacaagctg gagtacaact ataatcccca caacgtgtat
     5221 atcatggccg ataagcagaa gaacggcatc aaggtcaatt tcaagatccg ccacaatatc
     5281 gaggacggca gcgtgcagct ggccgatcac taccagcaga acaccccaat cggcgacgga
     5341 cccgtgctgc tgcctgataa tcactatctg tttacacagt ctgccctgag caaggacccc
     5401 aacgagaagc gggatcacat ggtgctgctg gagttcgtga ccgcagcagg catcacactg
     5461 ggcatggatg agctgtacaa gggcagcgcc ggcaccatgg cacaggacct gtccgagaag
     5521 gatctgctga agatggaggt ggagcagctg aagaaggagg tgaagaatac aagaatcccc
     5581 atcagcaagg ccggcaagga gatcaaggag tatgtggagg cccaggccgg caacgaccct
     5641 ttcctgaagg gcatcccaga ggataagaat ccctttaagg agaagggcgg ctgcatcatc
     5701 agctgagggc ccagatctcc tgaggctgca gtctagagga tcataatcag ccataccaca
     5761 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     5821 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     5881 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgccgac gtcaggtggc
     5941 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     6001 atgtatccgc tcatgagaca ataaccctga taaatataaa tgtacatatt atgatataga
     6061 tacaacgtat gcaatggcca atagccaata ttgatttatg ctatataacc aatgaataat
     6121 atggctaatg gccaatattg aagatctgtc gagccatgtg agcaaaaggc cagcaaaagg
     6181 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     6241 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6301 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6361 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6421 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6481 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6541 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6601 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6661 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6721 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6781 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6841 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6901 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6961 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     7021 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     7081 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     7141 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     7201 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     7261 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     7321 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7381 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7441 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7501 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7561 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     7621 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7681 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7741 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7801 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7861 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7921 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     7981 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     8041 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     8101 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     8161 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgt
//
