LOCUS       sequence-478493-        6838 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express NanoLuc-DGK_ 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 6838)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6838)
  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            "Unpublished"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-DGK_ Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6838
                     /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(3917..3935)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3972..4093
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3983..4002)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4037..4056
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4338..4695
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4338..4529
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4361..4381)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4546..4681
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4710..4757
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4789..5583
                     /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(4843..4862)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5453..5472
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5647..5695
                     /note="synthetic polyadenylation signal"
     rep_origin      5852..6397
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     6298..6317
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6514..6531
                     /label=L4440
                     /note="L4440 vector, forward primer"
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 ggcaaaggag aggggactga
     1621 tcagcccatc cgacttcgcc cagctgcaga agtacatgga gtattctacc aagaaggtga
     1681 gcgacgtgct gaagctgttt gaggatggcg agatggccaa gtacgtgcag ggcgacgcca
     1741 tcggctatga gggcttccag cagtttctga agatctacct ggaggtggat aatgtgcctc
     1801 ggcacctgtc tctggccctg ttccagagct ttgagaccgg ccactgcctg aacgagacca
     1861 atgtgacaaa ggacgtggtg tgcctgaatg acgtgagctg ttacttctct ctgctggagg
     1921 gcggcagacc agaggataag ctggagttca cctttaagct gtatgacaca gataggaacg
     1981 gcatcctgga cagctccgag gtggataaga tcatcctgca gatgatgcgc gtggccgagt
     2041 acctggactg ggatgtgagc gagctgcggc ccatcctgca ggagatgatg aaggagatcg
     2101 actatgatgg atctggaagc gtgtcccagg cagagtgggt gcgggcagga gccaccacag
     2161 tgcctctgct ggtgctgctg ggcctggaga tgaccctgaa ggacgatggc cagcacatgt
     2221 ggcggccaaa gagatttcct aggccagtgt attgcaacct gtgcgagtct agcatcggcc
     2281 tgggcaagca gggcctgtcc tgcaacctgt gcaagtacac cgtgcacgac cagtgcgcca
     2341 tgaaggccct gccttgtgag gtgtccacat atgccaagtc tcgcaaggac atcggcgtgc
     2401 agagccacgt gtgggtgcgg ggaggatgcg agtccggccg ctgcgatcgg tgtcagaaga
     2461 agatccgcat ctaccacagc ctgaccggcc tgcactgcgt gtggtgtcac ctggagatcc
     2521 acgacgattg tctgcaggca gtgggacacg agtgcgactg tggactgctg cgcgatcaca
     2581 tcctgccccc ttcctctatc tatccaagcg tgctggcatc cggacctgac aggaagaaca
     2641 gcaagacctc ccagaagaca atggacgatc tgaatctgtc taccagcgag gccctgagga
     2701 tcgatccagt gcctaacaca cacccactgc tggtgttcgt gaatcccaag tccggcggca
     2761 agcagggcca gagagtgctg tggaagttcc agtacatcct gaacccaagg caggtgttta
     2821 atctgctgaa ggacggcccc gagatcggcc tgagactgtt taaggacgtg cctgatagca
     2881 ggatcctggt gtgcggaggc gacggaaccg tgggctggat cctggagaca atcgataagg
     2941 ccaacctgcc cgtgctgcca ccagtggccg tgctgcctct gggcaccggc aatgatctgg
     3001 caagatgtct gagatgggga ggaggatacg agggacagaa cctggccaag atcctgaagg
     3061 acctggagat gtccaaggtg gtgcacatgg atagatggtc tgtggaagtg atcccacagc
     3121 agacagagga gaagtccgac ccagtgccct tccagatcat caacaattat ttttccatcg
     3181 gcgtggatgc ctctatcgcc caccgcttcc acatcatgcg ggagaagtac cccgagaagt
     3241 ttaactctag gatgaagaat aagctgtggt atttcgagtt tgccacctcc gagtctatct
     3301 tcagcacatg caagaagctg gaggagtccc tgaccgtgga gatctgtggc aagcccctgg
     3361 acctgtctaa tctgagcctg gagggcatcg ccgtgctgaa catcccaagc atgcacggcg
     3421 gctccaatct gtggggcgac acacggagac cccacggcga tatctacggc atcaaccagg
     3481 ccctgggagc aaccgccaaa gtgatcacag accctgatat cctgaagacc tgcgtgccag
     3541 acctgtctga taagagactg gaggtggtgg gactggaggg agcaatcgag atgggccaga
     3601 tctatacaaa gctgaagaac gcaggcaggc gcctggcaaa gtgcagcgag atcaccttcc
     3661 acaccacaaa gacactgccc atgcagatcg acggcgagcc ttggatgcag accccatgta
     3721 caatcaagat cacccacaag aaccagatgc ctatgctgat gggccctcca cccaggtcta
     3781 caaatttctt tggctttctg agcgtttaaa cgaattcggg ctcggtaccc ggggatcctc
     3841 tagagtcgac ctgcaggcat gcaagctgat ccggctgcta acaaagcccg aaaggaagct
     3901 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ggccgcttcg
     3961 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     4021 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     4081 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat
     4141 gtgggaggtt tttttaagca agtaaaacct ctacaaatgt ggtaaaatcg aattttaaca
     4201 aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     4261 ttcacaccgc atacgcggat ctgcgcagca ccatggcctg aaataacctc tgaaagagga
     4321 acttggttag gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     4381 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4441 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4501 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4561 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4621 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4681 ctaggctttt gcaaaaagct taattaactg ttgacaatta atcatcggca tagtatatcg
     4741 gcatagtata atacgacaag gtgaggaact aaacccagga ggcagatcat gattgaacaa
     4801 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4861 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4921 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     4981 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     5041 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     5101 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     5161 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     5221 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     5281 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     5341 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     5401 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5461 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5521 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5581 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgat
     5641 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg
     5701 atagcgataa ggatccgctc atgcgatgat aagctgtcaa acatgagaat tacaacttat
     5761 atcgtatggg gctgacttca ggtgctacat ttgaagagat aaattgcact gaaatctaga
     5821 aatattttat ctgattaata agatgatctt cttgagatcg ttttggtctg cgcgtaatct
     5881 cttgctctga aaacgaaaaa accgccttgc agggcggttt ttcgaaggtt ctctgagcta
     5941 ccaactcttt gaaccgaggt aactggcttg gaggagcgca gtcaccaaaa cttgtccttt
     6001 cagtttagcc ttaaccggcg catgacttca agactaactc ctctaaatca attaccagtg
     6061 gctgctgcca gtggtgcttt tgcatgtctt tccgggttgg actcaagacg atagttaccg
     6121 gataaggcgc agcggtcgga ctgaacgggg ggttcgtgca tacagtccag cttggagcga
     6181 actgcctacc cggaactgag tgtcaggcgt ggaatgagac aaacgcggcc ataacagcgg
     6241 aatgacaccg gtaaaccgaa aggcaggaac aggagagcgc acgagggagc cgccaggggg
     6301 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac cactgatttg agcgtcagat
     6361 ttcgtgatgc ttgtcagggg ggcggagcct atggaaaaac ggctttgccg cggccctctc
     6421 acttccctgt taagtatctt cctggcatct tccaggaaat ctccgccccg ttcgtaagcc
     6481 atttccgctc gccgcagtcg aacgaccgag cgtagcgagt cagtgagcga ggaagcggaa
     6541 tatatcctgt atcacatatt ctgctgacgc accggtgcag ccttttttct cctgccacat
     6601 gaagcacttc actgacaccc tcatcagtgc caacatagta agccagtata cactccgcta
     6661 gcgctgatgt ccggcggtgc ttttgccgtt acgcaccacc ccgtcatgag attacgcgtt
     6721 ttagcggtgc gtacaattaa gggattatgg taaatccact tactgtctgc cctcgtagcc
     6781 atcgagataa accgcagtac tccggccacg atgcgtccgg cgtagaggat cgagatct
//
