LOCUS       sequence-306388-        7039 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nanoluc complementation assay. Expression of G-alpha-13 protein 
            fused with the LgBiT(LgB)of the nanoluciferase inserted between 
            residues 106 and 107 of G-alpha-13. Addition of the HA epitope at N 
            term of G-alpha-13..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7039)
  AUTHORS   Laschet C, Dupuis N, Hanson J
  TITLE     A dynamic and screening-compatible nanoluciferase-based 
            complementation assay enables profiling of individual GPCR-G protein
            interactions.
  JOURNAL   J Biol Chem. 2019 Mar 15;294(11):4079-4090. doi: 
            10.1074/jbc.RA118.006231. Epub 2018 Dec 28.
  PUBMED    30593506
REFERENCE   2  (bases 1 to 7039)
  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: 
            "10.1074/jbc.RA118.006231"; journalName: "J Biol Chem"; date: 
            "2019-03-15- 15"; volume: "294"; issue: "11"; pages: "4079-4090"
COMMENT     Sequence Label: pcDNA3.1.G-alpha-13-LgB106
FEATURES             Location/Qualifiers
     source          1..7039
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             907..933
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2630..2647)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2636..2860
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2906..3334
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2993..3012)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3203..3224
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3343..3363)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3348..3677
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3528..3663
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3590..3609
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3744..4538
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3798..3817)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4408..4427
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4712..4833
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4749..4768)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4803..4822
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4882..4898)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4882..4898)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4895..4917)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4906..4922
                     /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(4930..4936,4937..4954,4955..4960))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4975..4996
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5113..5130)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5284..5872)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5364..5383)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6043..6834,6835..6903))
                     /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     6666..6685
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6904..7008)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 accatgtacc catacgatgt tccagattac gctgcggact tcctgccgtc gcggtccgtg
      961 ctgtccgtgt gcttccccgg ctgcctgctg acgagtggcg aggccgagca gcaacgcaag
     1021 tccaaggaga tcgacaaatg cctgtctcgg gaaaagacct atgtgaagcg gctggtgaag
     1081 atcctgctgc tgggcgcggg cgagagcggc aagtccacct tcctgaagca gatgcggatc
     1141 atccacgggc aggacttcga ccagcgcgcg cgcgaggagt tccgccccac catctacagc
     1201 aacgtgatca aaggtatgag ggtgctggtt gatgctcgag agaagcttga tatcagcgga
     1261 ggcggaggaa gcgtcttcac actcgaagat ttcgttgggg actgggaaca gacagccgcc
     1321 tacaacctgg accaagtcct tgaacaggga ggtgtgtcca gtttgctgca gaatctcgcc
     1381 gtgtccgtaa ctccgatcca aaggattgtc cggagcggtg aaaatgccct gaagatcgac
     1441 atccatgtca tcatcccgta tgaaggtctg agcgccgacc aaatggccca gatcgaagag
     1501 gtgtttaagg tggtgtaccc tgtggatgat catcacttta aggtgatcct gccctatggc
     1561 acactggtaa tcgacggggt tacgccgaac atgctgaact atttcggacg gccgtatgaa
     1621 ggcatcgccg tgttcgacgg caaaaagatc actgtaacag ggaccctgtg gaacggcaac
     1681 aaaattatcg acgagcgcct gatcaccccc gacggctcca tgctgttccg agtaaccatc
     1741 aacagtagcg gaggcggagg aagcccgcgg catattccct ggggagacaa ctcaaaccaa
     1801 caacatggag ataagatgat gtcgtttgat acccgggccc ccatggcagc ccaaggaatg
     1861 gtggaaacaa gggttttctt acaatatctt cctgctataa gagcattatg ggcagacagc
     1921 ggcatacaga atgcctatga ccggcgtcga gaatttcaac tgggtgaatc tgtaaaatat
     1981 ttcctggata acttggataa acttggagaa ccagattata ttccatcaca acaagatatt
     2041 ctgcttgcca gaagacccac caaaggcatc catgaatacg actttgaaat aaaaaatgtt
     2101 cctttcaaaa tggttgatgt aggtggtcag agatcagaaa ggaaacgttg gtttgaatgt
     2161 ttcgacagtg tgacatcaat acttttcctt gtttcctcaa gtgaatttga ccaggtgctt
     2221 atggaagatc gactgaccaa tcgccttaca gagtctctga acatttttga aacaatcgtc
     2281 aataaccggg ttttcagcaa tgtctccata attctgttct taaacaagac agacttgctt
     2341 gaggagaagg tgcaaattgt gagcatcaaa gactatttcc tagaatttga aggggatccc
     2401 cactgcttaa gagacgtcca aaaattcctg gtggaatgtt tccggaacaa acgccgggac
     2461 cagcaacaga agcccttata ccaccacttc accactgcta tcaacacgga gaacatccgc
     2521 cttgttttcc gtgacgtgaa ggatactatt ctgcatgaca acctcaagca gcttatgcta
     2581 cagtgagaat tcctcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg
     2641 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     2701 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     2761 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     2821 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     2881 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2941 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3001 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3061 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3121 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3181 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3241 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3301 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     3361 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3421 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3481 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3541 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3601 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3661 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     3721 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     3781 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     3841 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     3901 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     3961 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4021 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4081 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4141 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4201 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4261 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4321 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4381 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4441 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4501 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     4561 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4621 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4681 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     4741 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4801 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     4861 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4921 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4981 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5041 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5101 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5161 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5221 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5281 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5341 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5401 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5461 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5521 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5581 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5641 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5701 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5761 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5821 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5881 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5941 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6001 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6061 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6121 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6181 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6241 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6301 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6361 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6421 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6481 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6541 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6601 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6661 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6721 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6781 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6841 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6901 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6961 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7021 aaaagtgcca cctgacgtc
//
