LOCUS       sequence-222169-        6672 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding the nuclear localization of the chimeric 
            SNAP-NanoLuc fusion S-Luc in mammalian cells for BRET.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6672)
  AUTHORS   Hiblot J, Yu Q, Sabbadini MDB, Reymond L, Xue L, Schena A, Sallin O,
            Hill N, Griss R, Johnsson K
  TITLE     Luciferases with Tunable Emission Wavelengths.
  JOURNAL   Angew Chem Int Ed Engl. 2017 Nov 13;56(46):14556-14560. doi: 
            10.1002/anie.201708277. Epub 2017 Oct 9.
  PUBMED    28941028
REFERENCE   2  (bases 1 to 6672)
  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: 
            "10.1002/anie.201708277"; journalName: "Angew Chem Int Ed Engl"; 
            date: "2017-11-13- 13"; volume: "56"; issue: "46"; pages: 
            "14556-14560"
COMMENT     Sequence Label: pCDNA3.1-S-Luc_nucl
FEATURES             Location/Qualifiers
     source          1..6672
                     /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             914..955
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             956..976
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    1007..1031
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1139..1651
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             2129..2149
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2153..2173
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2177..2197
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(2220..2244)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2263..2280)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2269..2493
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2539..2967
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2626..2645)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2836..2857
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2976..2996)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2981..3310
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3161..3296
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3223..3242
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3377..4171
                     /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(3431..3450)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4041..4060
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4345..4466
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4382..4401)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4436..4455
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4515..4531)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4515..4531)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4528..4550)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4539..4555
                     /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(4563..4569,4570..4587,4588..4593))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4608..4629
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4746..4763)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4917..5505)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4997..5016)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5676..6467,6468..6536))
                     /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     6299..6318
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6537..6641)
                     /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 gctggctagt
      901 taagcttacc atgggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacggaaaa
      961 cctgtatttt cagggcccgg atccgtcgac gaattctgca gatatcacaa gtttgtacaa
     1021 aaaagcaggc ttcaccacca tgatggataa agactgcgaa atgaaacgca ctaccctgga
     1081 cagcccgctg ggtaaactgg aactgagcgg ttgcgagcag ggcctgcacg aaatcattat
     1141 ggtcttcaca ctcgaagatt tcgttgggga ctggcgacag acagccggct acaacctgga
     1201 ccaagtcctt gaacagggag gtgtgtccag tttgtttcag aatctcgggg tgtccgtaac
     1261 tccgatccaa aggattgtcc tgagcggtga aaatgggctg aagatcgaca tccatgtcat
     1321 catcccgtat gaaggtctga gcggcgacca aatgggccag atcgaaaaaa tttttaaggt
     1381 ggtgtaccct gtggatgatc atcactttaa ggtgatcctg cactatggca cactggtaat
     1441 cgacggggtt acgccgaaca tgatcgacta tttcggacgg ccgtatgaag gcatcgccgt
     1501 gttcgacggc aaaaagatca ctgtaacagg gaccctgtgg aacggcaaca aaattatcga
     1561 cgagcgcctg atcaaccccg acggctccct gctgttccga gtaaccatca acggagtgac
     1621 cggctggcgg ctgtgcgaac gcattctggc gttcctgggc aaaggtacga gcgctgcgga
     1681 tgcggtcgaa gtaccggctc cggctgcggt tctgggtggc ccggagccgc tgatgcaggc
     1741 taccgcatgg ctgaacgcat attttcacca gccagaggcg attgaagagt tccctgtacc
     1801 ggccctgcat cacccggtat ttcagcagga atcctttacc cgtcaggtac tgtggaaact
     1861 gctgaaagta gtgaaattcg gcgaggtgat ctcctattcc catctggcgg cgctggctgg
     1921 caacccagcg gcaactgcgg cggtgaaaac cgctctgtct ggtaatccgg tgccaattct
     1981 gatcccgtgc caccgtgtgg ttcagggcga cctggacgta ggtggctacg aaggcggcct
     2041 ggctgtgaaa gaatggctgc tggcgcacga aggtcaccgt ctgggtaaac caggcctggg
     2101 caagtccgga ctcagatctc gagctgatcc aaaaaagaag agaaaggtag atccaaaaaa
     2161 gaagagaaag gtagatccaa aaaagaagag aaaggtagga tccaccggat ctagatagga
     2221 cccagctttc ttgtacaaag tggtgataaa cccgctgatc agcctcgact gtgccttcta
     2281 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     2341 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     2401 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     2461 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     2521 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     2581 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     2641 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     2701 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     2761 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     2821 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     2881 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     2941 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     3001 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3061 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3121 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3181 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3241 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3301 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     3361 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     3421 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     3481 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     3541 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     3601 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     3661 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     3721 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     3781 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     3841 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     3901 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     3961 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     4021 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     4081 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     4141 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     4201 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     4261 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     4321 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     4381 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     4441 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     4501 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     4561 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     4621 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     4681 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     4741 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4801 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4861 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4921 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtta gaggtggcga
     4981 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5041 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5101 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5161 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5221 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5281 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5341 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5401 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5461 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5521 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5581 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5641 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5701 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     5761 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5821 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5881 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5941 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6001 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6061 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6121 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6181 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6241 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6301 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6361 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6421 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6481 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6541 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6601 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6661 ccacctgacg tc
//
