LOCUS       sequence-479796-        6578 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express NanoLuc-PLK1 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 6578)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6578)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoLuc-PLK1 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6578
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1065..1577
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(3521..3539)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3576..3697
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3587..3606)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3641..3660
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3942..4299
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3942..4133
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3965..3985)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4150..4285
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4314..4361
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4393..5187
                     /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(4447..4466)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5057..5076
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5251..5299
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5538..6126)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5618..5637)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6242..6525
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcgct caccatggtc ttcacactcg
     1081 aagatttcgt tggggactgg cgacagacag ccggctacaa cctggaccaa gtccttgaac
     1141 agggaggtgt gtccagtttg tttcagaatc tcggggtgtc cgtaactccg atccaaagga
     1201 ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca tgtcatcatc ccgtatgaag
     1261 gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt taaggtggtg taccctgtgg
     1321 atgatcatca ctttaaggtg atcctgcact atggcacact ggtaatcgac ggggttacgc
     1381 cgaacatgat cgactatttc ggacggccgt atgaaggcat cgccgtgttc gacggcaaaa
     1441 agatcactgt aacagggacc ctgtggaacg gcaacaaaat tatcgacgag cgcctgatca
     1501 accccgacgg ctccctgctg ttccgagtaa ccatcaacgg agtgaccggc tggcggctgt
     1561 gcgaacgcat tctggcgggc tcgagcggcg cgatcgccat gagtgctgca gtgactgcag
     1621 ggaagctggc acgggcaccg gccgaccctg ggaaagccgg ggtccccgga gttgcagctc
     1681 ccggagctcc ggcggcggct ccaccggcga aagagatccc ggaggtccta gtggacccac
     1741 gcagccggcg gcgctatgtg cggggccgct ttttgggcaa gggcggcttt gccaagtgct
     1801 tcgagatctc ggacgcggac accaaggagg tgttcgcggg caagattgtg cctaagtctc
     1861 tgctgctcaa gccgcaccag agggagaaga tgtccatgga aatatccatt caccgcagcc
     1921 tcgcccacca gcacgtcgta ggattccacg gctttttcga ggacaacgac ttcgtgttcg
     1981 tggtgttgga gctctgccgc cggaggtctc tcctggagct gcacaagagg aggaaagccc
     2041 tgactgagcc tgaggcccga tactacctac ggcaaattgt gcttggctgc cagtacctgc
     2101 accgaaaccg agttattcat cgagacctca agctgggcaa ccttttcctg aatgaagatc
     2161 tggaggtgaa aataggggat tttggactgg caaccaaagt cgaatatgac ggggagagga
     2221 agaagaccct gtgtgggact cctaattaca tagctcccga ggtgctgagc aagaaagggc
     2281 acagtttcga ggtggatgtg tggtccattg ggtgtatcat gtataccttg ttagtgggca
     2341 aaccaccttt tgagacttct tgcctaaaag agacctacct ccggatcaag aagaatgaat
     2401 acagtattcc caagcacatc aaccccgtgg ccgcctccct catccagaag atgcttcaga
     2461 cagatcccac tgcccgccca accattaacg agctgcttaa tgacgagttc tttacttctg
     2521 gctatatccc tgcccgtctc cccatcacct gcctgaccat tccaccaagg ttttcgattg
     2581 ctcccagcag cctggacccc agcaaccgga agcccctcac agtcctcaat aaaggcttgg
     2641 agaaccccct gcctgagcgt ccccgggaaa aagaagaacc agtggttcga gagacaggtg
     2701 aggtggtcga ctgccacctc agtgacatgc tgcagcagct gcacagtgtc aatgcctcca
     2761 agccctcgga gcgtgggctg gtcaggcaag aggaggctga ggatcctgcc tgcatcccca
     2821 tcttctgggt cagcaagtgg gtggactatt cggacaagta cggccttggg tatcagctct
     2881 gtgataacag cgtgggggtg ctcttcaatg actcaacacg cctcatcctc tacaatgatg
     2941 gtgacagcct gcagtacata gagcgtgacg gcactgagtc ctacctcacc gtgagttccc
     3001 atcccaactc cttgatgaag aagatcaccc tccttaaata tttccgcaat tacatgagcg
     3061 agcacttgct gaaggcaggt gccaacatca cgccgcgcga aggtgatgag ctcgcccggc
     3121 tgccctacct acggacctgg ttccgcaccc gcagcgccat catcctgcac ctcagcaacg
     3181 gcagcgtgca gatcaacttc ttccaggatc acaccaagct catcttgtgc ccactgatgg
     3241 cagccgtgac ctacatcgac gagaagcggg acttccgcac ataccgcctg agtctcctgg
     3301 aggagtacgg ctgctgcaag gagctggcca gccggctccg ctacgcccgc actatggtgg
     3361 acaagctgct gagctcacgc tcggccagca accgtctcaa ggcctccgtt taaacgaatt
     3421 cgggctcggt acccggggat cctctagagt cgacctgcag gcatgcaagc tgatccggct
     3481 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
     3541 taaccccttg gggcggccgc ttcgagcaga catgataaga tacattgatg agtttggaca
     3601 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     3661 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     3721 tatgtttcag gttcaggggg agatgtggga ggttttttta agcaagtaaa acctctacaa
     3781 atgtggtaaa atcgaatttt aacaaaatat taacgcttac aatttcctga tgcggtattt
     3841 tctccttacg catctgtgcg gtatttcaca ccgcatacgc ggatctgcgc agcaccatgg
     3901 cctgaaataa cctctgaaag aggaacttgg ttaggtacct tctgaggcgg aaagaaccag
     3961 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4021 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4081 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4141 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4201 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     4261 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttaatta actgttgaca
     4321 attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg aactaaaccc
     4381 aggaggcaga tcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4441 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4501 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4561 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4621 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4681 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4741 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4801 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4861 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4921 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4981 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5041 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5101 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5161 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5221 cgacgcccaa cctgccatca cgatggccgc aataaaatat ctttattttc attacatctg
     5281 tgtgttggtt ttttgtgtga atcgatagcg ataaggatcc tctttgcgct tgcgttttcc
     5341 cttgtccaga tagcccagta gctgacattc atccggggtc agcaccgttt ctgcggactg
     5401 gctttctacc cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5461 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5521 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5581 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5641 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5701 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5761 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5821 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5881 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5941 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     6001 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6061 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6121 tttcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6181 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttatag
     6241 tccggaaata caggaacgca cgctggatgg cccttcgctg ggatggtgaa accatgaaaa
     6301 atggcagctt cagtggatta agtgggggta atgtggcctg taccctctgg ttgcataggt
     6361 attcatacgg ttaaaattta tcaggcgcga ttgcggcagt ttttcgggtg gtttgttgcc
     6421 atttttacct gtctgctgcc gtgatcgcgc tgaacgcgtt ttagcggtgc gtacaattaa
     6481 gggattatgg taaatccact tactgtctgc cctcgtagcc atcgagataa accgcagtac
     6541 tccggccacg atgcgtccgg cgtagaggat cgagatct
//
