LOCUS       sequence-501951-        7425 bp    DNA     circular SYN 04-MAR-2026
DEFINITION  Express NanoLuc(R)-TCF4 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 7425)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7425)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 4, 2026 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            "Unpublished"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, TCF4
FEATURES             Location/Qualifiers
     source          1..7425
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    798..879
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     823..842
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2960..3472)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     promoter        complement(3511..3529)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3512..3529)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        3649..3952
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4107..4127
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        4160..4207
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4239..4898
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4305..4324)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(5070..5658)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5150..5169)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5784..5832
                     /note="synthetic polyadenylation signal"
     CDS             complement(5856..6650)
                     /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(5967..5986)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6577..6596
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(7028..7425,1..781))
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          join(7037..7425,1..550)
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(7060..7080)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 ctggtgccga ccgggccagc cttgccgccc agggcggggc gatacacggc ggcgcgaggc
       61 caggcaccag agcaggccgg ccagcttgag actacccccg tccgattctc ggtggccgcg
      121 ctcgcaggcc ccgcctcgcc gaacatgtgc gctgggacgc acgggccccg tcgccgcccg
      181 cggccccaaa aaccgaaata ccagtgtgca gatcttggcc cgcatttaca agactatctt
      241 gccagaaaaa aagcgtcgca gcaggtcatc aaaaatttta aatggctaga gacttatcga
      301 aagcagcgag acaggcgcga aggtgccacc agattcgcac gcggcggccc cagcgcccag
      361 gccaggcctc aactcaagca cgaggcgaag gggctcctta agcgcaaggc ctcgaactct
      421 cccacccact tccaacccga agctcgggat caagaatcac gtactgcagc caggtggaag
      481 taattcaagg cacgcaaggg ccataacccg taaagaggcc aggcccgcgg gaaccacaca
      541 cggcacttac ctgtgttctg gcggcaaacc cgttgcgaaa aagaacgttc acggcgacta
      601 ctgcacttat atacggttct cccccaccct cgggaaaaag gcggagccag tacacgacat
      661 cactttccca gtttaccccg cgccaccttc tctaggcacc ggttcaattg ccgacccctc
      721 cccccaactt ctcggggact gtgggcgatg tgcgctctgc ccactgacgg gcaccggagc
      781 caagacgcgt gacgtcggca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
      841 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
      901 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
      961 aaatgtggta tggctgatta tgatccgcgg ccgcaatacg tcgacgttat cagctgactt
     1021 cgtacgagag cctaggtgtt tctagaggat ccccgggtac cgagcccgaa ttcgtttaaa
     1081 cttctaaaga cttggtgacg agcgacagcg gctggggctg ggtcccggcc agggagctgt
     1141 gggaatgtaa ggaagaagtc gacggctgtg caattgatga ggagggggcg gcaggctgca
     1201 aagcggctgg gggcaggtcc agggccccgt tgggacagag ggcggaggcc ttgtgggtgg
     1261 cctcagcgag caggagggcg ggtggcggag gcatcatgga caggtgggcc agggggtcgg
     1321 gcttcaggga cagcgacaga ggctgggtct gctcagtctg tgacttggcg tctcggggag
     1381 gggagcctag caggttcggg gaggggggag gcgaatctag taagcttcca tctgaagagg
     1441 gtgggctgag gcagctgcct tcaccttgta tgtagcgaac gcactttttt tttctcctgc
     1501 aagggccgca ccagttattc tgttgatcaa ggccaaagcg cgctcggcat ttcttaggag
     1561 cgctcaggtc tgtaatcgga ggaagtgaaa ggcaaggatt taggaaacat tcgctgtgtt
     1621 cattggtctc tcccggctgc ttgtcccttt tcctcttctt cttctttcca tagttatccc
     1681 gcgcggacca gccggggtac agttgcatat gaagctgtcg ctccttccgg gccagctcgt
     1741 agtatttcgc ttgctcttct ctggacagtg catgccacct ccgcccaagg atctggttga
     1801 tggccgcgct ttctttcaac gtgcactcag ctacgacctt tgctctcatt tccttcatat
     1861 acaacatgaa tgcattaaga ggtttcttta tgtggggctt cttcttttct tcttcctttt
     1921 tggagtcctg atgctttgaa ctatggagtg agccgacatc actctgggac gattcctgtt
     1981 tgactgttgg tgtgactatg gccggatgcg gaatgcccgt cgtgtgtagc gtatgatgtg
     2041 gtgggaccat atggggaggg aacctggaca tggaagcatt gacggtcaga gctgtggggt
     2101 aggggtgtct gaatcctcct gtcgtgattg ggtacactgg ttgaccttgc tgtggtacta
     2161 accatcctag cggatggggg atttgtccta cggtgccagg cgatagtggg taatacgggg
     2221 atatatctgg agggtgcgga ggccgtggga ttcctgtttt ggggtctacg tcggctggta
     2281 agtgtggagg tgggtttccc ggcgtgaagt gttcattgct gtacgtgata agaggcgtga
     2341 gggggtggac atggtgaggg tgctgcacca ctggcacttt gttagagaca atgtgtgccg
     2401 gtgatgggga ccgggcatcc ttgagggctt gtctactctg gaggctccct gcctggacat
     2461 caagcagtgg ccatttcatc tggagatact gaactgcaat ctggtgttca atcgttttgt
     2521 acgcagagta atgtgtgctg ccggactgaa aatggagggt tcgggcggtg ggcgagagcg
     2581 atccgttggg gaggtagggg ctcgtcaggt cggggatcat gatgaagggg tagccgggat
     2641 acggtggccc cttaaagagc cctccatctt gcctcttggc cgcttcttcc aaactttccc
     2701 gggatttgtc tcggaaactt tcggagcgag gcggaggccg tctttccgcc tcggaatcgg
     2761 aggagctgtt ttgattcgtt tctgattcat tgactagaga cgatttgaca tcagctaaat
     2821 ccctctctgc cgaggagttt tcggagctct tctcctcctg ttcgccctcg tctttgaagg
     2881 aaatcagttc gtcgttggcg cctaggtcat cccctccacc gccgttcagc tgcggcatgg
     2941 cgatcgcgcc gctcgagccc gccagaatgc gttcgcacag ccgccagccg gtcactccgt
     3001 tgatggttac tcggaacagc agggagccgt cggggttgat caggcgctcg tcgataattt
     3061 tgttgccgtt ccacagggtc cctgttacag tgatcttttt gccgtcgaac acggcgatgc
     3121 cttcatacgg ccgtccgaaa tagtcgatca tgttcggcgt aaccccgtcg attaccagtg
     3181 tgccatagtg caggatcacc ttaaagtgat gatcatccac agggtacacc accttaaaaa
     3241 ttttttcgat ctggcccatt tggtcgccgc tcagaccttc atacgggatg atgacatgga
     3301 tgtcgatctt cagcccattt tcaccgctca ggacaatcct ttggatcgga gttacggaca
     3361 ccccgagatt ctgaaacaaa ctggacacac ctccctgttc aaggacttgg tccaggttgt
     3421 agccggctgt ctgtcgccag tccccaacga aatcttcgag tgtgaagacc atggtgagcg
     3481 ctagctgcaa aaagaacaag tagcttgtat tctatagtgt cacctaaatg ggcgatctga
     3541 cggttcacta aaccagctct gcttatatag gcctccctgg cgcgccacta gttattaata
     3601 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     3661 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     3721 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     3781 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtccgccccc
     3841 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttacg
     3901 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     3961 gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     4021 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     4081 atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac ggtgggaggt
     4141 caaatcgaat tctaatactg ttgacaatta atcatcggca tagtatatcg gcatagtata
     4201 atacgacaag gtgaggaact aaacccagga ggcagatcat ggagaaaaaa atcactggat
     4261 ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag
     4321 ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg
     4381 taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga
     4441 atgctcatcc ggaattccgt atggcaatga aagacggtga gctggtgata tgggatagtg
     4501 ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg
     4561 aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg
     4621 gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca
     4681 atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg
     4741 cccccgtttt cactatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg
     4801 cgattcaggt tcatcatgcc gtctgtgatg gcttccatgt cggcagaatg cttaatgaat
     4861 tacaacagta ctgcgatgag tggcagggcg gggcgtaagc gggactctgg ggttcgaaat
     4921 gaccgaccaa gcgacgccca accggtatca gctcactcaa aggcggtaat acggttatcc
     4981 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5041 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5101 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5161 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5221 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5281 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5341 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5401 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5461 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     5521 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5581 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5641 agaaaaaaag gatttcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5701 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5761 atccttttat agtccggaat cgtcacacaa aaaaccaaca cacagatgta atgaaaataa
     5821 agatatttta ttgcggccat cgtgatggct agagttcaga agaactcgtc aagaaggcga
     5881 tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca
     5941 gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag
     6001 cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc
     6061 atgatattcg gcaagcaggc atcgccgtgg gtcacgacga gatcctcgcc gtcgggcatg
     6121 cgcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga
     6181 tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc
     6241 gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca
     6301 gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc
     6361 acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg
     6421 caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcctg cagttcattc
     6481 agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg
     6541 aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc
     6601 tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat ggtggcatcg
     6661 attcccggtc aacatccagt acacctccgg cacaccggct tccccaaggg cgccacgctc
     6721 tcccaccaca acatcctcaa caacggctac tgggtgggcc gcacggtcgg ctacacggag
     6781 caggaccggg tgtgtctgcc ggtgcccttc taccactgct tcggcatggt catgggcaac
     6841 ttgggcgcca cctcccacgc cgcctgcatc gtcatcccgg ccccgtcctt cgagccggcg
     6901 gcgacgctgg aggcggtgca gcgggagagg tgcacgtccc tgtacggcgt ccccacgaag
     6961 ccgaattcca ccacactgga ctagatgcat gctcgacagt gtgatggata tctgcagaat
     7021 tcggctttca cgacacctga aatggaagaa aaaaactttg aaccactgtc tgaggcttga
     7081 gaatgaacca agatccaaac tcaaaaaggg caaattccaa ggagaattac atcaagtgcc
     7141 aagctggcct aacttcagtc tccacccact cagtgtgggg aaactccatc gcataaaacc
     7201 cctcccccca acctaaagac gacgtactcc aaaagctcga gaactaatcg aggtgcctgg
     7261 acggcgcccg gtactccgtg gagtcacatg aagcgacggc tgaggacgga aaggcccttt
     7321 tcctttgtgt gggtgactca cccgcccgct ctcccgagcg ccgcgtcctc cattttgagc
     7381 tgcctgcagc agggccggga agcggccatc tttccgctca cgcaa
//
