LOCUS       sequence-474575-        9846 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express BCL9-NanoLuc(R) Fusion Protein in Mammalian Cells under a 
            CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9846)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9846)
  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: 
            "Unpublished"
COMMENT     Sequence Label: NanoBRET Assay Vector, BCL9
FEATURES             Location/Qualifiers
     source          1..9846
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    17..98
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     42..61
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4600..5112)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     promoter        complement(5151..5169)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5152..5169)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        5289..5592
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     5747..5767
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        5800..5847
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5879..6538
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5945..5964)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(6710..7298)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6790..6809)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    7424..7472
                     /note="synthetic polyadenylation signal"
     CDS             complement(7496..8290)
                     /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(7607..7626)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     8217..8236
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(8668..9846)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          8677..9615
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(8700..8720)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 aagacgcgtg acgtcggcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
       61 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
      121 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
      181 aatgtggtat ggctgattat gatccgcggc cgcaatacgt cgacgttatc agctgacttc
      241 gtacgagagc ctaggtgttt ctagaggatc cccgggtacc gagcccgaat tcgtttaaac
      301 aaacatcatg tttcctgggt tgccaggtcc ttggctaaat ccacccatgc ccacgtcagc
      361 agccatgccc cggggcctca tctgtggggg gatcatgatg ttctgttggg gtcccatcat
      421 gccctgcatg gacatcatca tgcctggaga gcccacaggc ccagggtggg tgtagagccc
      481 ggcaggcccc cgatccttgc caggaatcat gcccacggca gcagctggat tgctcatcag
      541 ggtgggctgg ccgggcattg tagattgtgc tggtgacatc atccgatggt gaggtcccat
      601 catgccttgt tggagaaagg ctggtggtct catggggttg tggcctggca tggatggagc
      661 tgtaccaaga gggatgtccg gagttcccac tggccctgga cctcccatgc caggtaatgc
      721 tagtcccatt ctgggggctt gttcgcccat catcccctgc atgtgtgaaa acccaggtcc
      781 aggaccctgg ggctgtttac ggcctggaac ttcccctcga gggaaatatt gcagcgtctg
      841 gctgggcttc tcagatggaa taatgcggga tagatcaaac tcaggtattc cggtggctcc
      901 aggtcggatg acctcctgca gatctgggtc tgtaaacacc gaaggcatgc tgttccccag
      961 gacagtgaag gagtcaggac ccccggggcc tccaggcttg caaagtgctg catctgctga
     1021 actttggggc aggttgctgg ggcggccaag ggggccttct cctgggaaac ccatccctcc
     1081 tgggaagctg ccctgccccc cactggggcc attgtgaggg aatggaacct gctgtggggg
     1141 agactgtact ggagggaagc cctgggggaa gcccatccgt ccttgaggta ccatcggtgg
     1201 ctcctgggac ccatgcccca tgataggatt gtgtgacatc aagccaggcc ccattgggac
     1261 cccatgagga ggtatgttgg gtcccacggc atttggagag ggcatctgat tggagtgaga
     1321 aagtggctgg gtcattccca ttgggctgag ggttggcatc ggaaccacgg ggtttggacc
     1381 tgaaattcga ggattctgtg tattaatgcc cattcctggc atattattca ttgatggcaa
     1441 gttgggagaa cgagctggag gggagtcgtc atctgagctg gccacagtct tgatagcatc
     1501 atggtataac ggggtggaac tgggcattgc aaacttggac attcgagaca tcataataga
     1561 gagtgggttc tgggaaaggg ttggctctgg aggcatggta taaggtgtac tagagggaag
     1621 acttccaggg atattcacag aggcaggctg gctggctgta ggaggtgggg ggccacctga
     1681 ctctacattt cccagcatgg ctggggaggc catggtgagg ggtgctttat ggtttggagg
     1741 gatcccagga ctctgaaggg gaggttttgg agaagaggtc catccaggtg acggggcagg
     1801 aagtgatgga gacttgaggt ggacaggtga agcagcagca gaccccaaaa caggggggga
     1861 cttaatggaa gcagcagcag ctgggcccgc cagcatgcct gccagctgcg atggagtctg
     1921 gggggacttg aggttccccg agggcgagcc cagcattggt gactggactt ggtgcatcgt
     1981 gggagacttc agagggttaa tgcctgggga atgcacctgg ctgcctgcca cagatatatc
     2041 caagggcttc cgccccaggc cgcgctgaac tggaggagct gtgttgaggc tggtggggtt
     2101 actggaaggg ttgagaggta gtggtggcat atgactgagc cggctgttag tcctctggtc
     2161 gggcccaatt ggttctctga gattccgcaa gccactgttg ctgcctggac cctgagacat
     2221 gggaaggaat ggtctggggc ccatgccata ctcctgctgg gggtgctcac caaatggcag
     2281 tggcaccatc ttctgctgag caggcagcat gtctgagcca cctgggcgta atttcagcat
     2341 ctcctcaggg cccgccccag cctctctcat cttctgaggt atcatctgag agttggatcc
     2401 catgttgaca tttagattga catctccctt catcccacta ggaaccatcc caaactcaag
     2461 ttcccgacca gggcccatct gtgggggaat tccttttgga aagtcacccc tagaaggact
     2521 cagagggccc tcaactggta ttcgagggaa aatggggtta ttcccaggct ccatgtggcg
     2581 ctgggagcct ggaatcatcc tgttcatctc catgctgggc ctgatgcctt ccatggccat
     2641 ccctgggggg agacccagct gtttctctgc cagctgctgc tgaaacatct cttcagacaa
     2701 tccttggggg tttgggaagc gttcccctcg gccaggaccg ctgaaaatgc cctggccagg
     2761 aggaaaattt cgaccatctg ggatttttgg cacatcatct ggccaactga ctccagaaag
     2821 acctggtcta gatgcagggt tggggacatt cggcccttcc atttcagagt ttatcatgcc
     2881 tgcaaatcca gggaggcgca tctggctccc tggcatgttg gggtggggag ccatgcccct
     2941 cgggggcaga gaatgtggca tgttgatacc atcagaaaat ggctctgtac ccccaggtgc
     3001 ccagccttca ctaggggtca tctggtatgg aggggggggt cctcggacca ctccccgagg
     3061 cccgtgctga tggaccatca tgtcctggag ggaacactgc tggacaacca cttgttcctg
     3121 cttcctcctc ttctcttcat aaaactcctg ctgcagtttc agccacgcta tctgctcggg
     3181 agtcatatgg tcaaggtggt cgggtcctat ggtcccagac tgggagttca tagaaggtgg
     3241 aaccatttca tctggagaaa agggtacatc tctatgtcct tgagggccaa atggagctcc
     3301 cacgtctgtc cggggcccag gtcccttacc taggctttgg gattgggcca tcatggcctg
     3361 tattggccct tctggttttt tctgaggccc atctaatacc ccaggattct gctgcggtcc
     3421 cccactttgt gctcctgtga attctttctc atcaggaaaa agcatgcgct ggatatctct
     3481 gagagtttgt aaggagcgct cccggtgctc cagctgctcc tgagataggc catcgggatt
     3541 ctctcccagt gtggggggct cgccactgga cactggtgga ggcggagggg ctttgggatc
     3601 tgctgaagag ctattgctcc cctgggagac aggggtcact gccctattgt tgggagttga
     3661 gttgggccca gtaccatctg ggggcagtgg agtggagctg gcaggacttc ctacagaagg
     3721 aatcagtttg ttttctaccc caggactctc ccggtccagg ggacgtgggg gtgcggcagg
     3781 cttgggtgct ggtgccggaa tgggtggagt tggctgcagt ctggtattct gggaagaatt
     3841 ctggtcctgg ttggccggag ctgggggctg ttgtgggaga ggtttcggat cattccgaag
     3901 ggcagatatc tgtgtgttca gaggcgctgt gcttctctct gtcttgttgt tagaaatgtt
     3961 ctggatgtgg aaagagacga tagtttcaac ctggcccttc aaaacagctt ctgcagcttt
     4021 attggccatc tcagtagaaa acacgtacac cactttggct ggagtcttct gagcaggtgt
     4081 gggctctgtg gcagtagttt ggccatggga gggggtagaa gacctggggg ctgtagcatt
     4141 tgatggggtc atcgagtgtg gtgtgtgctg ggaatcctgg gactttatgt ggtcagcaga
     4201 attacattct ttaatgtcag agtcatcgtt tggagtccca ggatctctct gatcaaagga
     4261 gtcggcggaa atacttcgct cccttttccc cttgcccttg gcaccatttc cagccccatt
     4321 cttcagcccc atgctcccac ctgggccagg gagtgcttta ggggtatggc ccccactctt
     4381 ggagtcacag ggggatggct gggattggct ggctgagccc ccctgtttac cctgattgga
     4441 gaatttggaa tccagctggg ggtttccaga tggggacatc actgtagggg gacggaccat
     4501 cacctcctgc tttgacttag ggctactctg tgtgtttcct gatggagagc tcctcacttt
     4561 agggttactg gaatgcatgg cgatcgcgcc gctcgagccc gccagaatgc gttcgcacag
     4621 ccgccagccg gtcactccgt tgatggttac tcggaacagc agggagccgt cggggttgat
     4681 caggcgctcg tcgataattt tgttgccgtt ccacagggtc cctgttacag tgatcttttt
     4741 gccgtcgaac acggcgatgc cttcatacgg ccgtccgaaa tagtcgatca tgttcggcgt
     4801 aaccccgtcg attaccagtg tgccatagtg caggatcacc ttaaagtgat gatcatccac
     4861 agggtacacc accttaaaaa ttttttcgat ctggcccatt tggtcgccgc tcagaccttc
     4921 atacgggatg atgacatgga tgtcgatctt cagcccattt tcaccgctca ggacaatcct
     4981 ttggatcgga gttacggaca ccccgagatt ctgaaacaaa ctggacacac ctccctgttc
     5041 aaggacttgg tccaggttgt agccggctgt ctgtcgccag tccccaacga aatcttcgag
     5101 tgtgaagacc atggtgagcg ctagctgcaa aaagaacaag tagcttgtat tctatagtgt
     5161 cacctaaatg ggcgatctga cggttcacta aaccagctct gcttatatag gcctccctgg
     5221 cgcgccacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     5281 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     5341 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     5401 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     5461 catatgccaa gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     5521 gcccagtaca tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc
     5581 gctattacca tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac
     5641 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     5701 aatcaacggg actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt
     5761 aggcgtgtac ggtgggaggt caaatcgaat tctaatactg ttgacaatta atcatcggca
     5821 tagtatatcg gcatagtata atacgacaag gtgaggaact aaacccagga ggcagatcat
     5881 ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca
     5941 ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat
     6001 tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca
     6061 cattcttgcc cgcctgatga atgctcatcc ggaattccgt atggcaatga aagacggtga
     6121 gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac
     6181 gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc
     6241 gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa
     6301 tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc
     6361 caatatggac aacttcttcg cccccgtttt cactatgggc aaatattata cgcaaggcga
     6421 caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtctgtgatg gcttccatgt
     6481 cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaagc
     6541 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca accggtatca gctcactcaa
     6601 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6661 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6721 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6781 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6841 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6901 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6961 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     7021 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     7081 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     7141 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     7201 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     7261 gcaagcagca gattacgcgc agaaaaaaag gatttcaaga agatcctttg atcttttcta
     7321 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     7381 caaaaaggat cttcacctag atccttttat agtccggaat cgtcacacaa aaaaccaaca
     7441 cacagatgta atgaaaataa agatatttta ttgcggccat cgtgatggct agagttcaga
     7501 agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt
     7561 aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag
     7621 ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag
     7681 aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccgtgg gtcacgacga
     7741 gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct ggcgcgagcc
     7801 cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg
     7861 ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat
     7921 gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg
     7981 acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga
     8041 caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg
     8101 cctcgtcctg cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc
     8161 gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc
     8221 agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt
     8281 gttcaatcat ggtggcatcg attcccggtc aacatccagt acacctccgg cacaccggct
     8341 tccccaaggg cgccacgctc tcccaccaca acatcctcaa caacggctac tgggtgggcc
     8401 gcacggtcgg ctacacggag caggaccggg tgtgtctgcc ggtgcccttc taccactgct
     8461 tcggcatggt catgggcaac ttgggcgcca cctcccacgc cgcctgcatc gtcatcccgg
     8521 ccccgtcctt cgagccggcg gcgacgctgg aggcggtgca gcgggagagg tgcacgtccc
     8581 tgtacggcgt ccccacgaag ccgaattcca ccacactgga ctagatgcat gctcgacagt
     8641 gtgatggata tctgcagaat tcggctttca cgacacctga aatggaagaa aaaaactttg
     8701 aaccactgtc tgaggcttga gaatgaacca agatccaaac tcaaaaaggg caaattccaa
     8761 ggagaattac atcaagtgcc aagctggcct aacttcagtc tccacccact cagtgtgggg
     8821 aaactccatc gcataaaacc cctcccccca acctaaagac gacgtactcc aaaagctcga
     8881 gaactaatcg aggtgcctgg acggcgcccg gtactccgtg gagtcacatg aagcgacggc
     8941 tgaggacgga aaggcccttt tcctttgtgt gggtgactca cccgcccgct ctcccgagcg
     9001 ccgcgtcctc cattttgagc tgcctgcagc agggccggga agcggccatc tttccgctca
     9061 cgcaactggt gccgaccggg ccagccttgc cgcccagggc ggggcgatac acggcggcgc
     9121 gaggccaggc accagagcag gccggccagc ttgagactac ccccgtccga ttctcggtgg
     9181 ccgcgctcgc aggccccgcc tcgccgaaca tgtgcgctgg gacgcacggg ccccgtcgcc
     9241 gcccgcggcc ccaaaaaccg aaataccagt gtgcagatct tggcccgcat ttacaagact
     9301 atcttgccag aaaaaaagcg tcgcagcagg tcatcaaaaa ttttaaatgg ctagagactt
     9361 atcgaaagca gcgagacagg cgcgaaggtg ccaccagatt cgcacgcggc ggccccagcg
     9421 cccaggccag gcctcaactc aagcacgagg cgaaggggct ccttaagcgc aaggcctcga
     9481 actctcccac ccacttccaa cccgaagctc gggatcaaga atcacgtact gcagccaggt
     9541 ggaagtaatt caaggcacgc aagggccata acccgtaaag aggccaggcc cgcgggaacc
     9601 acacacggca cttacctgtg ttctggcggc aaacccgttg cgaaaaagaa cgttcacggc
     9661 gactactgca cttatatacg gttctccccc accctcggga aaaaggcgga gccagtacac
     9721 gacatcactt tcccagttta ccccgcgcca ccttctctag gcaccggttc aattgccgac
     9781 ccctcccccc aacttctcgg ggactgtggg cgatgtgcgc tctgcccact gacgggcacc
     9841 ggagcc
//
