LOCUS       sequence-410275-        7056 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Part of REGA-SIGN, a NanoBRET-based G protein biosensor. It encodes 
            GNA13 tagged with NanoLuciferase and is meant to be paired with 
            Ggamma1 labeled with LSS-mKATE2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7056)
  AUTHORS   Boon K, Vanalken N, Meyen E, Schols D, Van Loy T
  TITLE     REGA-SIGN: Development of a Novel Set of NanoBRET-Based G Protein 
            Biosensors.
  JOURNAL   Biosensors (Basel). 2023 Jul 28;13(8):767. doi: 
            10.3390/bios13080767.
  PUBMED    37622853
REFERENCE   2  (bases 1 to 7056)
  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: "Biosensors 
            (Basel)."; date: "2023-07-28"; volume: "13(8):767. doi"; pages: " 
            10.3390/bios13080767"
COMMENT     Sequence Label: pcDNA3.1(+)G-alpha-13-125-NLuc
FEATURES             Location/Qualifiers
     source          1..7056
                     /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             1319..1828
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2647..2664)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2653..2877
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2923..3351
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3010..3029)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3220..3241
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3360..3380)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3365..3694
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3545..3680
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3607..3626
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3761..4555
                     /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(3815..3834)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4425..4444
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4729..4850
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4766..4785)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4820..4839
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4899..4915)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4899..4915)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4912..4934)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4923..4939
                     /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(4947..4953,4954..4971,4972..4977))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4992..5013
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5130..5147)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5301..5889)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5381..5400)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6060..6851,6852..6920))
                     /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     6683..6702
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6921..7025)
                     /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 gtttaaactt aagcttggta ccaccatggc ggacttcctg ccgtcgcggt ccgtgctgtc
      961 cgtgtgcttc cccggctgcc tgctgacgag tggcgaggcc gagcagcaac gcaagtccaa
     1021 ggagatcgac aaatgcctgt ctcgggaaaa gacctatgtg aagcggctgg tgaagatcct
     1081 gctgctgggc gcgggcgaga gcggcaagtc caccttcctg aagcagatgc ggatcatcca
     1141 cgggcaggac ttcgaccagc gcgcgcgcga ggagttccgc cccaccatct acagcaacgt
     1201 gatcaaaggt atgagggtgc tggttgatgc tcgagagaag cttcatattc cctggggaga
     1261 caactcaaac caacaacatg gagataagat gatgtcgttt agcggcggcg gcggcagcgt
     1321 cttcacactc gaagatttcg ttggggactg gcgacagaca gccggctaca acctggacca
     1381 agtccttgaa cagggaggtg tgtccagttt gtttcagaat ctcggggtgt ccgtaactcc
     1441 gatccaaagg attgtcctga gcggtgaaaa tgggctgaag atcgacatcc atgtcatcat
     1501 cccgtatgaa ggtctgagcg gcgaccaaat gggccagatc gaaaaaattt ttaaggtggt
     1561 gtaccctgtg gatgatcatc actttaaggt gatcctgcac tatggcacac tggtaatcga
     1621 cggggttacg ccgaacatga tcgactattt cggacggccg tatgaaggca tcgccgtgtt
     1681 cgacggcaaa aagatcactg taacagggac cctgtggaac ggcaacaaaa ttatcgacga
     1741 gcgcctgatc aaccccgacg gctccctgct gttccgagta accatcaacg gagtgaccgg
     1801 ctggcggctg tgcgaacgca ttctggcgag cggcggcggc ggcagcgata cccgggcccc
     1861 catggcagcc caaggaatgg tggaaacaag ggttttctta caatatcttc ctgctataag
     1921 agcattatgg gcagacagcg gcatacagaa tgcctatgac cggcgtcgag aatttcaact
     1981 gggtgaatct gtaaaatatt tcctggataa cttggataaa cttggagaac cagattatat
     2041 tccatcacaa caagatattc tgcttgccag aagacccacc aaaggcatcc atgaatacga
     2101 ctttgaaata aaaaatgttc ctttcaaaat ggttgatgta ggtggtcaga gatcagaaag
     2161 gaaacgttgg tttgaatgtt tcgacagtgt gacatcaata cttttccttg tttcctcaag
     2221 tgaatttgac caggtgctta tggaagatcg actgaccaat cgccttacag agtctctgaa
     2281 catttttgaa acaatcgtca ataaccgggt tttcagcaat gtctccataa ttctgttctt
     2341 aaacaagaca gacttgcttg aggagaaggt gcaaattgtg agcatcaaag actatttcct
     2401 agaatttgaa ggggatcccc actgcttaag agacgtccaa aaattcctgg tggaatgttt
     2461 ccggaacaaa cgccgggacc agcaacagaa gcccttatac caccacttca ccactgctat
     2521 caacacggag aacatccgcc ttgttttccg tgacgtgaag gatactattc tgcatgacaa
     2581 cctcaagcag cttatgctac agtgatctag agatctctag agggcccgtt taaacccgct
     2641 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     2701 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     2761 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     2821 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     2881 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     2941 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3001 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3061 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3121 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3181 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3241 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     3301 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     3361 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3421 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3481 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3541 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3601 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     3661 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     3721 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3781 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3841 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3901 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     3961 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     4021 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     4081 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     4141 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4201 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     4261 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     4321 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4381 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4441 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4501 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4561 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4621 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4681 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     4741 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4801 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     4861 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     4921 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4981 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     5041 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5101 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5161 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5221 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5281 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5341 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5401 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5461 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5521 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5581 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5641 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5701 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5761 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5821 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5881 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5941 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6001 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6061 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6121 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6181 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6241 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6301 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6361 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6421 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6481 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6541 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6601 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6661 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6721 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6781 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6841 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6901 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6961 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7021 ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
