LOCUS       sequence-470734-        7426 bp    DNA     circular SYN 15-DEC-2025
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 7426)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7426)
  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            "Unpublished"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, TCF4
FEATURES             Location/Qualifiers
     source          1..7426
                     /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(2179..2691)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     promoter        complement(2730..2748)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2731..2748)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        2868..3171
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3326..3346
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3379..3426
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3458..4117
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3524..3543)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(4289..4877)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4369..4388)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5003..5051
                     /note="synthetic polyadenylation signal"
     CDS             complement(5075..5869)
                     /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(5186..5205)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5796..5815
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(6248..7426)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6257..7195
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(6280..6300)
                     /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 ttctaaagac ttggtgacga gcgacagcgg ctggggctgg gtcccggcca gggagctgtg
      361 ggaatgtaag gaagaagtcg acggctgtgc aattgatgag gagggggcgg caggctgcaa
      421 agcggctggg ggcaggtcca gggccccgtt gggacagagg gcggaggcct tgtgggtggc
      481 ctcagcgagc aggagggcgg gtggcggagg catcatggac aggtgggcca gggggtcggg
      541 cttcagggac agcgacagag gctgggtctg ctcagtctgt gacttggcgt ctcggggagg
      601 ggagcctagc aggttcgggg aggggggagg cgaatctagt aagcttccat ctgaagaggg
      661 tgggctgagg cagctgcctt caccttgtat gtagcgaacg cacttttttt ttctcctgca
      721 agggccgcac cagttattct gttgatcaag gccaaagcgc gctcggcatt tcttaggagc
      781 gctcaggtct gtaatcggag gaagtgaaag gcaaggattt aggaaacatt cgctgtgttc
      841 attggtctct cccggctgct tgtccctttt cctcttcttc ttctttccat agttatcccg
      901 cgcggaccag ccggggtaca gttgcatatg aagctgtcgc tccttccggg ccagctcgta
      961 gtatttcgct tgctcttctc tggacagtgc atgccacctc cgcccaagga tctggttgat
     1021 ggccgcgctt tctttcaacg tgcactcagc tacgaccttt gctctcattt ccttcatata
     1081 caacatgaat gcattaagag gtttctttat gtggggcttc ttcttttctt cttccttttt
     1141 ggagtcctga tgctttgaac tatggagtga gccgacatca ctctgggacg attcctgttt
     1201 gactgttggt gtgactatgg ccggatgcgg aatgcccgtc gtgtgtagcg tatgatgtgg
     1261 tgggaccata tggggaggga acctggacat ggaagcattg acggtcagag ctgtggggta
     1321 ggggtgtctg aatcctcctg tcgtgattgg gtacactggt tgaccttgct gtggtactaa
     1381 ccatcctagc ggatggggga tttgtcctac ggtgccaggc gatagtgggt aatacgggga
     1441 tatatctgga gggtgcggag gccgtgggat tcctgttttg gggtctacgt cggctggtaa
     1501 gtgtggaggt gggtttcccg gcgtgaagtg ttcattgctg tacgtgataa gaggcgtgag
     1561 ggggtggaca tggtgagggt gctgcaccac tggcactttg ttagagacaa tgtgtgccgg
     1621 tgatggggac cgggcatcct tgagggcttg tctactctgg aggctccctg cctggacatc
     1681 aagcagtggc catttcatct ggagatactg aactgcaatc tggtgttcaa tcgttttgta
     1741 cgcagagtaa tgtgtgctgc cggactgaaa atggagggtt cgggcggtgg gcgagagcga
     1801 tccgttgggg aggtaggggc tcgtcaggtc ggggatcatg atgaaggggt agccgggata
     1861 cggtggcccc ttaaagagcc ctccatcttg cctcttggcc gcttcttcca aactttcccg
     1921 ggatttgtct cggaaacttt cggagcgagg cggaggccgt ctttccgcct cggaatcgga
     1981 ggagctgttt tgattcgttt ctgattcatt gactagagac gatttgacat cagctaaatc
     2041 cctctctgcc gaggagtttt cggagctctt ctcctcctgt tcgccctcgt ctttgaagga
     2101 aatcagttcg tcgttggcgc ctaggtcatc ccctccaccg ccgttcagct gcggcatggc
     2161 gatcgcgccg ctcgagcccg ccagaatgcg ttcgcacagc cgccagccgg tcactccgtt
     2221 gatggttact cggaacagca gggagccgtc ggggttgatc aggcgctcgt cgataatttt
     2281 gttgccgttc cacagggtcc ctgttacagt gatctttttg ccgtcgaaca cggcgatgcc
     2341 ttcatacggc cgtccgaaat agtcgatcat gttcggcgta accccgtcga ttaccagtgt
     2401 gccatagtgc aggatcacct taaagtgatg atcatccaca gggtacacca ccttaaaaat
     2461 tttttcgatc tggcccattt ggtcgccgct cagaccttca tacgggatga tgacatggat
     2521 gtcgatcttc agcccatttt caccgctcag gacaatcctt tggatcggag ttacggacac
     2581 cccgagattc tgaaacaaac tggacacacc tccctgttca aggacttggt ccaggttgta
     2641 gccggctgtc tgtcgccagt ccccaacgaa atcttcgagt gtgaagacca tggtgagcgc
     2701 tagctgcaaa aagaacaagt agcttgtatt ctatagtgtc acctaaatgg gcgatctgac
     2761 ggttcactaa accagctctg cttatatagg cctccctggc gcgccactag ttattaatag
     2821 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     2881 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     2941 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     3001 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct
     3061 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg
     3121 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     3181 ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     3241 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     3301 tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     3361 aaatcgaatt ctaatactgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     3421 tacgacaagg tgaggaacta aacccaggag gcagatcatg gagaaaaaaa tcactggata
     3481 taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt
     3541 tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt
     3601 aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa
     3661 tgctcatccg gaattccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt
     3721 tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga
     3781 ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg
     3841 tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa
     3901 tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc
     3961 ccccgttttc actatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc
     4021 gattcaggtt catcatgccg tctgtgatgg cttccatgtc ggcagaatgc ttaatgaatt
     4081 acaacagtac tgcgatgagt ggcagggcgg ggcgtaagcg ggactctggg gttcgaaatg
     4141 accgaccaag cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata cggttatcca
     4201 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4261 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4321 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4381 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4441 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4501 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4561 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4621 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4681 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     4741 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4801 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4861 gaaaaaaagg atttcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4921 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4981 tccttttata gtccggaatc gtcacacaaa aaaccaacac acagatgtaa tgaaaataaa
     5041 gatattttat tgcggccatc gtgatggcta gagttcagaa gaactcgtca agaaggcgat
     5101 agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag
     5161 cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc
     5221 ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca
     5281 tgatattcgg caagcaggca tcgccgtggg tcacgacgag atcctcgccg tcgggcatgc
     5341 gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat
     5401 catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg
     5461 cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag
     5521 ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca
     5581 cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc
     5641 aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca
     5701 gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga
     5761 acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct
     5821 ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg gtggcatcga
     5881 ttcccggtca acatccagta cacctccggc accaccggct tccccaaggg cgccacgctc
     5941 tcccaccaca acatcctcaa caacggctac tgggtgggcc gcacggtcgg ctacacggag
     6001 caggaccggg tgtgtctgcc ggtgcccttc taccactgct tcggcatggt catgggcaac
     6061 ttgggcgcca cctcccacgc cgcctgcatc gtcatcccgg ccccgtcctt cgagccggcg
     6121 gcgacgctgg aggcggtgca gcgggagagg tgcacgtccc tgtacggcgt ccccacgaag
     6181 ccgaattcca ccacactgga ctagatgcat gctcgacagt gtgatggata tctgcagaat
     6241 tcggctttca cgacacctga aatggaagaa aaaaactttg aaccactgtc tgaggcttga
     6301 gaatgaacca agatccaaac tcaaaaaggg caaattccaa ggagaattac atcaagtgcc
     6361 aagctggcct aacttcagtc tccacccact cagtgtgggg aaactccatc gcataaaacc
     6421 cctcccccca acctaaagac gacgtactcc aaaagctcga gaactaatcg aggtgcctgg
     6481 acggcgcccg gtactccgtg gagtcacatg aagcgacggc tgaggacgga aaggcccttt
     6541 tcctttgtgt gggtgactca cccgcccgct ctcccgagcg ccgcgtcctc cattttgagc
     6601 tgcctgcagc agggccggga agcggccatc tttccgctca cgcaactggt gccgaccggg
     6661 ccagccttgc cgcccagggc ggggcgatac acggcggcgc gaggccaggc accagagcag
     6721 gccggccagc ttgagactac ccccgtccga ttctcggtgg ccgcgctcgc aggccccgcc
     6781 tcgccgaaca tgtgcgctgg gacgcacggg ccccgtcgcc gcccgcggcc ccaaaaaccg
     6841 aaataccagt gtgcagatct tggcccgcat ttacaagact atcttgccag aaaaaaagcg
     6901 tcgcagcagg tcatcaaaaa ttttaaatgg ctagagactt atcgaaagca gcgagacagg
     6961 cgcgaaggtg ccaccagatt cgcacgcggc ggccccagcg cccaggccag gcctcaactc
     7021 aagcacgagg cgaaggggct ccttaagcgc aaggcctcga actctcccac ccacttccaa
     7081 cccgaagctc gggatcaaga atcacgtact gcagccaggt ggaagtaatt caaggcacgc
     7141 aagggccata acccgtaaag aggccaggcc cgcgggaacc acacacggca cttacctgtg
     7201 ttctggcggc aaacccgttg cgaaaaagaa cgttcacggc gactactgca cttatatacg
     7261 gttctccccc accctcggga aaaaggcgga gccagtacac gacatcactt tcccagttta
     7321 ccccgcgcca ccttctctag gcaccggttc aattgccgac ccctcccccc aacttctcgg
     7381 ggactgtggg cgatgtgcgc tctgcccact gacgggcacc ggagcc
//
