LOCUS TRUPATH Triple (bidirectional) GasS 8203 bp DNA circular SYN 25-AUG-2024
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 Aug 25, 2024 from SnapGene Server 7.0.3
https://www.biofeng.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."
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(6974..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
//
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