LOCUS       sequence-222526-        6936 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding the cytosolic localization of the chimeric 
            HaloTag7-NanoLucCP fusion H-Luc in mammalian cells for BRET.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6936)
  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 6936)
  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-H-Luc_cyto
FEATURES             Location/Qualifiers
     source          1..6936
                     /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"
     protein_bind    complement(2484..2508)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2527..2544)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2533..2757
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2803..3231
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2890..2909)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3100..3121
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3240..3260)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3245..3574
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3425..3560
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3487..3506
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3641..4435
                     /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(3695..3714)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4305..4324
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4609..4730
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4646..4665)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4700..4719
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4779..4795)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4779..4795)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4792..4814)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4803..4819
                     /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(4827..4833,4834..4851,4852..4857))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4872..4893
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5010..5027)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5181..5769)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5261..5280)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5940..6731,6732..6800))
                     /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     6563..6582
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6801..6905)
                     /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 tgggatccga aatcggtact ggctttccat tcgaccccca
     1081 ttatgtggaa gtcctgggcg agcgcatgca ctacgtcgat gttggtccgc gcgatggcac
     1141 ccctgtgctg ttcctgcacg gtaacccgac ctcctcctac gtgtggcgca acatcatccc
     1201 gcatgttgca ccgacccatc gctgcattgc tccagacctg atcggtatgg gcaaatccga
     1261 caaaccagac ctgggttatt tcttcgacga ccacgtccgc ttcatggatg ccttcatcga
     1321 agccctgggt ctggaagagg tcgtcctggt cattcacgac tggggctccg ctctgggttt
     1381 ccactgggcc aagcgcaatc cagagcgcgt caaaggtatt gcatttatgg agttcatccg
     1441 ccctatcccg acctgggacg aatggccaga atttgcccgc gagaccttcc aggccttccg
     1501 caccagcggc gaccaaatgg gccagatcga aaaaattttt aaggtggtgt accctgtgga
     1561 tgatcatcac tttaaggtga tcctgcacta tggcacactg gtaatcgacg gggttacgcc
     1621 gaacatgatc gactatttcg gacggccgta tgaaggcatc gccgtgttcg acggcaaaaa
     1681 gatcactgta acagggaccc tgtggaacgg caacaaaatt atcgacgagc gcctgatcaa
     1741 ccccgacggc tccctgctgt tccgagtaac catcaacgga gtgaccggct ggcggctgtg
     1801 cgaacgcatt ctggcgggtg gcaccggtgg ctctggcggt accggtggct ctatggtctt
     1861 cacactcgaa gatttcgttg gggactggcg acagacagcc ggctacaacc tggaccaagt
     1921 ccttgaacag ggaggtgtgt ccagtttgtt tcagaatctc ggggtgtccg taactccgat
     1981 ccaaaggatt gtcctgagcg gtgaaaatgg gctgaagatc gacatccatg tcatcatccc
     2041 gtatgaaggt ctggacgtcg gccgcaagct gatcatcgat cagaacgttt ttatcgaggg
     2101 tacgctgccg atgggtgtcg tccgcccgct gactgaagtc gagatggacc attaccgcga
     2161 gccgttcctg aatcctgttg accgcgagcc actgtggcgc ttcccaaacg agctgccaat
     2221 cgccggtgag ccagcgaaca tcgtcgcgct ggtcgaagaa tacatggact ggctgcacca
     2281 gtcccctgtc ccgaagctgc tgttctgggg caccccaggc gttctgatcc caccggccga
     2341 agccgctcgc ctggccaaaa gcctgcctaa ctgcaaggct gtggacatcg gcccgggtct
     2401 gaatctgctg caagaagaca acccggacct gatcggcagc gagatcgcgc gctggctgtc
     2461 tactctggag atttccggtt aggacccagc tttcttgtac aaagtggtga taaacccgct
     2521 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     2581 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     2641 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     2701 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     2761 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     2821 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     2881 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2941 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3001 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3061 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3121 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     3181 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     3241 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3301 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3361 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3421 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3481 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     3541 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     3601 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3661 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3721 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3781 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     3841 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3901 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3961 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     4021 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4081 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     4141 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     4201 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4261 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4321 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4381 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4441 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4501 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4561 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     4621 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4681 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     4741 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     4801 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4861 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4921 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4981 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5041 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5101 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5161 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5221 cgacgctcaa gttagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5281 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5341 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5401 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5461 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5521 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5581 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5641 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5701 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5761 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5821 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5881 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5941 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6001 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6061 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6121 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6181 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6241 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6301 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6361 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6421 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6481 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6541 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6601 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6661 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6721 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6781 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6841 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6901 ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
