LOCUS       sequence-475033-        6544 bp    DNA     circular SYN 09-JUN-2025
DEFINITION  Express WDR5-NanoLuc 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 6544)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6544)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 9, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: WDR5-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6544
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1558..2352
                     /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(1612..1631)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2222..2241
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2376..2424
                     /note="synthetic polyadenylation signal"
     rep_origin      2550..3138
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3039..3058
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3310..3969)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3884..3903
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4001..4048)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     primer_bind     complement(4081..4101)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4256..4559
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4679..4697
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4679..4696
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             5758..6270
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     polyA_signal    6447..6528
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6484..6503)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg caggcagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgaa agccgaattc tgcagatatc
     1201 catcacactg tcgagcatgc atctagtcca gtgtggtgga attcggcttc gtggggacgc
     1261 cgtacaggga cgtgcacctc tcccgctgca ccgcctccag cgtcgccgcc ggctcgaagg
     1321 acggggccgg gatgacgatg caggcggcgt gggaggtggc gcccaagttg cccatgacca
     1381 tgccgaagca gtggtagaag ggcaccggca gacacacccg gtcctgctcc gtgtagccga
     1441 ccgtgcggcc cacccagtag ccgttgttga ggatgttgtg gtgggagagc gtggcgccct
     1501 tggggaagcc ggtggtgccg gaggtgtact ggatgttgac cgggaatcga tgccaccatg
     1561 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     1621 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     1681 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     1741 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     1801 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     1861 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     1921 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     1981 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     2041 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     2101 gaggatctcg tcgtgaccca cggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     2161 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     2221 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     2281 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     2341 gagttcttct gaactctagc catcacgatg gccgcaataa aatatcttta ttttcattac
     2401 atctgtgtgt tggttttttg tgtgacgatt ccggactata aaaggatcta ggtgaagatc
     2461 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2521 gaccccgtag aaaagatcaa aggatcttct tgaaatcctt tttttctgcg cgtaatctgc
     2581 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2641 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     2701 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2761 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2821 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2881 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2941 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3001 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3061 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3121 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3181 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3241 accgcctttg agtgagctga taccggttgg gcgtcgcttg gtcggtcatt tcgaacccca
     3301 gagtcccgct tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat
     3361 tctgccgaca tggaagccat cacagacggc atgatgaacc tgaatcgcca gcggcatcag
     3421 caccttgtcg ccttgcgtat aatatttgcc catagtgaaa acgggggcga agaagttgtc
     3481 catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa
     3541 aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac
     3601 atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga
     3661 tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat
     3721 caccagctca ccgtctttca ttgccatacg gaattccgga tgagcattca tcaggcgggc
     3781 aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa
     3841 ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc
     3901 ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt
     3961 tttctccatg atctgcctcc tgggtttagt tcctcacctt gtcgtattat actatgccga
     4021 tatactatgc cgatgattaa ttgtcaacag tattagaatt cgatttgacc tcccaccgta
     4081 cacgcctacc gcccatttgc gtcaacgggg cggggttatt acgacatttt ggaaagtccc
     4141 gttgattttg gtgccaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc
     4201 cccgtgagtc aaaccgctat ccacgcccat tggtgtactg ccaaaaccgc atcaccatgg
     4261 taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccgt aaggtcatgt
     4321 actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcggacttg
     4381 gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt
     4441 gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca
     4501 atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc
     4561 ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat taataactag
     4621 tggcgcgcca gggaggccta tataagcaga gctggtttag tgaaccgtca gatcgcccat
     4681 ttaggtgaca ctatagaata caagctactt gttctttttg cagctagcga tcgccatggc
     4741 gacggaggag aagaagcccg agaccgaggc cgccagagca cagccaaccc cttcgtcatc
     4801 cgccactcag agcaagccta cacctgtgaa gccaaactat gctctaaagt tcacccttgc
     4861 tggccacacc aaagcagtgt cctccgtgaa attcagcccg aatggagagt ggctggcaag
     4921 ttcatctgct gataaactta ttaaaatttg gggcgcgtat gatgggaaat ttgagaaaac
     4981 catatctggt cacaagctgg gaatatccga tgtagcctgg tcgtcagatt ctaaccttct
     5041 tgtttctgcc tcagatgaca aaaccttgaa gatatgggac gtgagctcgg gcaagtgtct
     5101 gaaaaccctg aagggacaca gtaattatgt cttttgctgc aacttcaatc cccagtccaa
     5161 ccttattgtc tcaggatcct ttgacgaaag cgtgaggata tgggatgtga aaacagggaa
     5221 gtgcctcaag actttgccag ctcactcgga tccagtctcg gccgttcatt ttaatcgtga
     5281 tggatccttg atagtttcaa gtagctatga tggtctctgt cgcatctggg acaccgcctc
     5341 aggccagtgc ctgaagacgc tcatcgatga cgacaacccc cccgtgtctt ttgtgaagtt
     5401 ctccccgaac ggcaaataca tcctggccgc cacgctggac aacactctga agctctggga
     5461 ctacagcaag gggaagtgcc tgaagacgta cactggccac aagaatgaga aatactgcat
     5521 atttgccaat ttctctgtta ctggtgggaa gtggattgtg tctggctcag aggataacct
     5581 tgtttacatc tggaaccttc agacgaaaga gattgtacag aaactacaag gccacacaga
     5641 tgtcgtgatc tcaacagctt gtcacccaac agaaaacatc atcgcctctg ctgcgctaga
     5701 aaatgacaaa acaattaaac tgtggaagag tgactgcgag ctctcggctc gagcggcgtc
     5761 ttcacactcg aagatttcgt tggggactgg cgacagacag ccggctacaa cctggaccaa
     5821 gtccttgaac agggaggtgt gtccagtttg tttcagaatc tcggggtgtc cgtaactccg
     5881 atccaaagga ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca tgtcatcatc
     5941 ccgtatgaag gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt taaggtggtg
     6001 taccctgtgg atgatcatca ctttaaggtg atcctgcact atggcacact ggtaatcgac
     6061 ggggttacgc cgaacatgat cgactatttc ggacggccgt atgaaggcat cgccgtgttc
     6121 gacggcaaaa agatcactgt aacagggacc ctgtggaacg gcaacaaaat tatcgacgag
     6181 cgcctgatca accccgacgg ctccctgctg ttccgagtaa ccatcaacgg agtgaccggc
     6241 tggcggctgt gcgaacgcat tctggcgtaa ggccgcgact ctagaaacac ctaggctctc
     6301 gtacgaagtc agctgataac gtcgacgtat tgcggccgcg gatcataatc agccatacca
     6361 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     6421 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     6481 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgccg acgtcacgcg
     6541 tctt
//
