LOCUS       sequence-479836-        7394 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express DGKz-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 7394)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7394)
  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            "Unpublished"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: DGKz-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..7394
                     /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"
     regulatory      2922..2931
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3873..4385
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(4473..4491)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4528..4649
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4539..4558)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4593..4612
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4894..5251
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4894..5085
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4917..4937)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5102..5237
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5266..5313
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5345..6139
                     /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(5399..5418)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6009..6028
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6203..6251
                     /note="synthetic polyadenylation signal"
     rep_origin      6408..6953
                     /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     6854..6873
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7070..7087
                     /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 ggctagcgat cgccatggag ccgcgggacg
     1081 gtagccccga ggcccggagc agcgactccg agtcggcttc cgcctcgtcc agcggctccg
     1141 agcgcgacgc cggtcccgag ccggacaagg cgccgcggcg actcaacaag cggcgcttcc
     1201 cggggctgcg gctcttcggg cacaggaaag ccatcacgaa gtcgggcctc cagcacctgg
     1261 ccccccctcc gcccacccct ggggccccgt gcagcgagtc agagcggcag atccggagta
     1321 cagtggactg gagcgagtca gcgacatatg gggagcacat ctggttcgag accaacgtgt
     1381 ccggggactt ctgctacgtt ggggagcagt actgtgtagc caggatgctg cagaagtcag
     1441 tgtctcgaag aaagtgcgca gcctgcaaga ttgtggtgca cacgccctgc atcgagcagc
     1501 tggagaagat aaatttccgc tgtaagccgt ccttccgtga atcaggctcc aggaatgtcc
     1561 gcgagccaac ctttgtacgg caccactggg tacacagacg acgccaggac ggcaagtgtc
     1621 ggcactgtgg gaagggattc cagcagaagt tcaccttcca cagcaaggag attgtggcca
     1681 tcagctgctc gtggtgcaag caggcatacc acagcaaggt gtcctgcttc atgctgcagc
     1741 agatcgagga gccgtgctcg ctgggggtcc acgcagccgt ggtcatcccg cccacctgga
     1801 tcctccgcgc ccggaggccc cagaatactc tgaaagcaag caagaagaag aagagggcat
     1861 ccttcaagag gaagtccagc aagaaagggc ctgaggaggg ccgctggaga cccttcatca
     1921 tcaggcccac cccctccccg ctcatgaagc ccctgctggt gtttgtgaac cccaagagtg
     1981 ggggcaacca gggtgcaaag atcatccagt ctttcctctg gtatctcaat ccccgacaag
     2041 tcttcgacct gagccaggga gggcccaagg aggcgctgga gatgtaccgc aaagtgcaca
     2101 acctgcggat cctggcgtgc gggggcgacg gcacggtggg ctggatcctc tccaccctgg
     2161 accagctacg cctgaagccg ccaccccctg ttgccatcct gcccctgggt actggcaacg
     2221 acttggcccg aaccctcaac tggggtgggg gctacacaga tgagcctgtg tccaagatcc
     2281 tctcccacgt ggaggagggg aacgtggtac agctggaccg ctgggacctc cacgctgagc
     2341 ccaaccccga ggcagggcct gaggaccgag atgaaggcgc caccgaccgg ttgcccctgg
     2401 atgtcttcaa caactacttc agcctgggct ttgacgccca cgtcaccctg gagttccacg
     2461 agtctcgaga ggccaaccca gagaaattca acagccgctt tcggaataag atgttctacg
     2521 ccgggacagc tttctctgac ttcctgatgg gcagctccaa ggacctggcc aagcacatcc
     2581 gagtggtgtg tgatggaatg gacttgactc ccaagatcca ggacctgaaa cccaagtgtg
     2641 ttgttttcct gaacatcccc aggtactgtg cgggcaccat gccctggggc caccctgggg
     2701 agcaccacga ctttgagccc cagcggcatg acgacggcta cctcgaggtc attggcttca
     2761 ccatgacgtc gttggccgcg ctgcaggtgg gcggacacgg cgagcggctg acgcagtgtc
     2821 gcgaggtggt gctcaccaca tccaaggcca tcccggtgca ggtggatggc gagccctgca
     2881 agcttgcagc ctcacgcatc cgcatcgccc tgcgcaacca ggccaccatg gtgcagaagg
     2941 ccaagcggcg gagcgccgcc cccctgcaca gcgaccagca gccggtgcca gagcagttgc
     3001 gcatccaggt gagtcgcgtc agcatgcacg actatgaggc cctgcactac gacaaggagc
     3061 agctcaagga ggcctctgtg ccgctgggca ctgtggtggt cccaggagac agtgacctag
     3121 agctctgccg tgcccacatt gagagactcc agcaggagcc cgatggtgct ggagccaagt
     3181 ccccgacatg ccagaaactg tcccccaagt ggtgcttcct ggacgccacc actgccagcc
     3241 gcttctacag gatcgaccga gcccaggagc acctcaacta tgtgactgag atcgcacagg
     3301 atgagattta tatcctggac cctgagctgc tgggggcatc ggcccggcct gacctcccaa
     3361 cccccacttc ccctctcccc acctcaccct gctcacccac gccccggtca ctgcaagggg
     3421 atgctgcacc ccctcaaggt gaagagctga ttgaggctgc caagaggaac gacttctgta
     3481 agctccagga gctgcaccga gctgggggcg acctcatgca ccgagacgag cagagtcgca
     3541 cgctcctgca ccacgcagtc agcactggca gcaaggatgt ggtccgctac ctgctggacc
     3601 acgccccccc agagatcctt gatgcggtgg aggaaaacgg ggagacctgt ttgcaccaag
     3661 cagcggccct gggccagcgc accatctgcc actacatcgt ggaggccggg gcctcgctca
     3721 tgaagacaga ccagcagggc gacactcccc ggcagcgggc tgagaaggct caggacaccg
     3781 agctggccgc ctacctggag aaccggcagc actaccagat gatccagcgg gaggaccagg
     3841 agacggctgt ggtttctctc ggctcgagcg gcgtcttcac actcgaagat ttcgttgggg
     3901 actggcgaca gacagccggc tacaacctgg accaagtcct tgaacaggga ggtgtgtcca
     3961 gtttgtttca gaatctcggg gtgtccgtaa ctccgatcca aaggattgtc ctgagcggtg
     4021 aaaatgggct gaagatcgac atccatgtca tcatcccgta tgaaggtctg agcggcgacc
     4081 aaatgggcca gatcgaaaaa atttttaagg tggtgtaccc tgtggatgat catcacttta
     4141 aggtgatcct gcactatggc acactggtaa tcgacggggt tacgccgaac atgatcgact
     4201 atttcggacg gccgtatgaa ggcatcgccg tgttcgacgg caaaaagatc actgtaacag
     4261 ggaccctgtg gaacggcaac aaaattatcg acgagcgcct gatcaacccc gacggctccc
     4321 tgctgttccg agtaaccatc aacggagtga ccggctggcg gctgtgcgaa cgcattctgg
     4381 cgtaaggccg cgactctaga gtcgacctgc aggcatgcaa gctgatccgg ctgctaacaa
     4441 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     4501 tggggcggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     4561 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     4621 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     4681 aggttcaggg ggagatgtgg gaggtttttt taagcaagta aaacctctac aaatgtggta
     4741 aaatcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta
     4801 cgcatctgtg cggtatttca caccgcatac gcggatctgc gcagcaccat ggcctgaaat
     4861 aacctctgaa agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa
     4921 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4981 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     5041 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     5101 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     5161 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     5221 aggctttttt ggaggcctag gcttttgcaa aaagcttaat taactgttga caattaatca
     5281 tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac ccaggaggca
     5341 gatcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     5401 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     5461 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     5521 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     5581 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     5641 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     5701 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     5761 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     5821 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     5881 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     5941 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     6001 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     6061 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     6121 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     6181 aacctgccat cacgatggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg
     6241 ttttttgtgt gaatcgatag cgataaggat ccgctcatgc gatgataagc tgtcaaacat
     6301 gagaattaca acttatatcg tatggggctg acttcaggtg ctacatttga agagataaat
     6361 tgcactgaaa tctagaaata ttttatctga ttaataagat gatcttcttg agatcgtttt
     6421 ggtctgcgcg taatctcttg ctctgaaaac gaaaaaaccg ccttgcaggg cggtttttcg
     6481 aaggttctct gagctaccaa ctctttgaac cgaggtaact ggcttggagg agcgcagtca
     6541 ccaaaacttg tcctttcagt ttagccttaa ccggcgcatg acttcaagac taactcctct
     6601 aaatcaatta ccagtggctg ctgccagtgg tgcttttgca tgtctttccg ggttggactc
     6661 aagacgatag ttaccggata aggcgcagcg gtcggactga acggggggtt cgtgcataca
     6721 gtccagcttg gagcgaactg cctacccgga actgagtgtc aggcgtggaa tgagacaaac
     6781 gcggccataa cagcggaatg acaccggtaa accgaaaggc aggaacagga gagcgcacga
     6841 gggagccgcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccaccact
     6901 gatttgagcg tcagatttcg tgatgcttgt caggggggcg gagcctatgg aaaaacggct
     6961 ttgccgcggc cctctcactt ccctgttaag tatcttcctg gcatcttcca ggaaatctcc
     7021 gccccgttcg taagccattt ccgctcgccg cagtcgaacg accgagcgta gcgagtcagt
     7081 gagcgaggaa gcggaatata tcctgtatca catattctgc tgacgcaccg gtgcagcctt
     7141 ttttctcctg ccacatgaag cacttcactg acaccctcat cagtgccaac atagtaagcc
     7201 agtatacact ccgctagcgc tgatgtccgg cggtgctttt gccgttacgc accaccccgt
     7261 catgagatta cgcgttttag cggtgcgtac aattaaggga ttatggtaaa tccacttact
     7321 gtctgccctc gtagccatcg agataaaccg cagtactccg gccacgatgc gtccggcgta
     7381 gaggatcgag atct
//
