LOCUS       sequence-479769-        7420 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express NanoLuc-DGKz 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 7420)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7420)
  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            "Unpublished"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-DGKz Fusion Vector
FEATURES             Location/Qualifiers
     source          1..7420
                     /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"
     regulatory      3456..3465
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4499..4517)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4554..4675
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4565..4584)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4619..4638
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4920..5277
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4920..5111
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4943..4963)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5128..5263
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5292..5339
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5371..6165
                     /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(5425..5444)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6035..6054
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6229..6277
                     /note="synthetic polyadenylation signal"
     rep_origin      6434..6979
                     /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     6880..6899
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7096..7113
                     /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 ggagccgcgg gacggtagcc
     1621 ccgaggcccg gagcagcgac tccgagtcgg cttccgcctc gtccagcggc tccgagcgcg
     1681 acgccggtcc cgagccggac aaggcgccgc ggcgactcaa caagcggcgc ttcccggggc
     1741 tgcggctctt cgggcacagg aaagccatca cgaagtcggg cctccagcac ctggcccccc
     1801 ctccgcccac ccctggggcc ccgtgcagcg agtcagagcg gcagatccgg agtacagtgg
     1861 actggagcga gtcagcgaca tatggggagc acatctggtt cgagaccaac gtgtccgggg
     1921 acttctgcta cgttggggag cagtactgtg tagccaggat gctgcagaag tcagtgtctc
     1981 gaagaaagtg cgcagcctgc aagattgtgg tgcacacgcc ctgcatcgag cagctggaga
     2041 agataaattt ccgctgtaag ccgtccttcc gtgaatcagg ctccaggaat gtccgcgagc
     2101 caacctttgt acggcaccac tgggtacaca gacgacgcca ggacggcaag tgtcggcact
     2161 gtgggaaggg attccagcag aagttcacct tccacagcaa ggagattgtg gccatcagct
     2221 gctcgtggtg caagcaggca taccacagca aggtgtcctg cttcatgctg cagcagatcg
     2281 aggagccgtg ctcgctgggg gtccacgcag ccgtggtcat cccgcccacc tggatcctcc
     2341 gcgcccggag gccccagaat actctgaaag caagcaagaa gaagaagagg gcatccttca
     2401 agaggaagtc cagcaagaaa gggcctgagg agggccgctg gagacccttc atcatcaggc
     2461 ccaccccctc cccgctcatg aagcccctgc tggtgtttgt gaaccccaag agtgggggca
     2521 accagggtgc aaagatcatc cagtctttcc tctggtatct caatccccga caagtcttcg
     2581 acctgagcca gggagggccc aaggaggcgc tggagatgta ccgcaaagtg cacaacctgc
     2641 ggatcctggc gtgcgggggc gacggcacgg tgggctggat cctctccacc ctggaccagc
     2701 tacgcctgaa gccgccaccc cctgttgcca tcctgcccct gggtactggc aacgacttgg
     2761 cccgaaccct caactggggt gggggctaca cagatgagcc tgtgtccaag atcctctccc
     2821 acgtggagga ggggaacgtg gtacagctgg accgctggga cctccacgct gagcccaacc
     2881 ccgaggcagg gcctgaggac cgagatgaag gcgccaccga ccggttgccc ctggatgtct
     2941 tcaacaacta cttcagcctg ggctttgacg cccacgtcac cctggagttc cacgagtctc
     3001 gagaggccaa cccagagaaa ttcaacagcc gctttcggaa taagatgttc tacgccggga
     3061 cagctttctc tgacttcctg atgggcagct ccaaggacct ggccaagcac atccgagtgg
     3121 tgtgtgatgg aatggacttg actcccaaga tccaggacct gaaacccaag tgtgttgttt
     3181 tcctgaacat ccccaggtac tgtgcgggca ccatgccctg gggccaccct ggggagcacc
     3241 acgactttga gccccagcgg catgacgacg gctacctcga ggtcattggc ttcaccatga
     3301 cgtcgttggc cgcgctgcag gtgggcggac acggcgagcg gctgacgcag tgtcgcgagg
     3361 tggtgctcac cacatccaag gccatcccgg tgcaggtgga tggcgagccc tgcaagcttg
     3421 cagcctcacg catccgcatc gccctgcgca accaggccac catggtgcag aaggccaagc
     3481 ggcggagcgc cgcccccctg cacagcgacc agcagccggt gccagagcag ttgcgcatcc
     3541 aggtgagtcg cgtcagcatg cacgactatg aggccctgca ctacgacaag gagcagctca
     3601 aggaggcctc tgtgccgctg ggcactgtgg tggtcccagg agacagtgac ctagagctct
     3661 gccgtgccca cattgagaga ctccagcagg agcccgatgg tgctggagcc aagtccccga
     3721 catgccagaa actgtccccc aagtggtgct tcctggacgc caccactgcc agccgcttct
     3781 acaggatcga ccgagcccag gagcacctca actatgtgac tgagatcgca caggatgaga
     3841 tttatatcct ggaccctgag ctgctggggg catcggcccg gcctgacctc ccaaccccca
     3901 cttcccctct ccccacctca ccctgctcac ccacgccccg gtcactgcaa ggggatgctg
     3961 caccccctca aggtgaagag ctgattgagg ctgccaagag gaacgacttc tgtaagctcc
     4021 aggagctgca ccgagctggg ggcgacctca tgcaccgaga cgagcagagt cgcacgctcc
     4081 tgcaccacgc agtcagcact ggcagcaagg atgtggtccg ctacctgctg gaccacgccc
     4141 ccccagagat ccttgatgcg gtggaggaaa acggggagac ctgtttgcac caagcagcgg
     4201 ccctgggcca gcgcaccatc tgccactaca tcgtggaggc cggggcctcg ctcatgaaga
     4261 cagaccagca gggcgacact ccccggcagc gggctgagaa ggctcaggac accgagctgg
     4321 ccgcctacct ggagaaccgg cagcactacc agatgatcca gcgggaggac caggagacgg
     4381 ctgtggttta aacgaattcg ggctcggtac ccggggatcc tctagagtcg acctgcaggc
     4441 atgcaagctg atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc
     4501 gctgagcaat aactagcata accccttggg gcggccgctt cgagcagaca tgataagata
     4561 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     4621 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     4681 caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg tttttttaag
     4741 caagtaaaac ctctacaaat gtggtaaaat cgaattttaa caaaatatta acgcttacaa
     4801 tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgcgg
     4861 atctgcgcag caccatggcc tgaaataacc tctgaaagag gaacttggtt aggtaccttc
     4921 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4981 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     5041 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     5101 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     5161 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     5221 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     5281 cttaattaac tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca
     5341 aggtgaggaa ctaaacccag gaggcagatc atgattgaac aagatggatt gcacgcaggt
     5401 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     5461 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     5521 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     5581 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     5641 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     5701 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     5761 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     5821 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     5881 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     5941 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     6001 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     6061 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     6121 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     6181 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg atggccgcaa taaaatatct
     6241 ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagcgat aaggatccgc
     6301 tcatgcgatg ataagctgtc aaacatgaga attacaactt atatcgtatg gggctgactt
     6361 caggtgctac atttgaagag ataaattgca ctgaaatcta gaaatatttt atctgattaa
     6421 taagatgatc ttcttgagat cgttttggtc tgcgcgtaat ctcttgctct gaaaacgaaa
     6481 aaaccgcctt gcagggcggt ttttcgaagg ttctctgagc taccaactct ttgaaccgag
     6541 gtaactggct tggaggagcg cagtcaccaa aacttgtcct ttcagtttag ccttaaccgg
     6601 cgcatgactt caagactaac tcctctaaat caattaccag tggctgctgc cagtggtgct
     6661 tttgcatgtc tttccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6721 gactgaacgg ggggttcgtg catacagtcc agcttggagc gaactgccta cccggaactg
     6781 agtgtcaggc gtggaatgag acaaacgcgg ccataacagc ggaatgacac cggtaaaccg
     6841 aaaggcagga acaggagagc gcacgaggga gccgccaggg ggaaacgcct ggtatcttta
     6901 tagtcctgtc gggtttcgcc accactgatt tgagcgtcag atttcgtgat gcttgtcagg
     6961 ggggcggagc ctatggaaaa acggctttgc cgcggccctc tcacttccct gttaagtatc
     7021 ttcctggcat cttccaggaa atctccgccc cgttcgtaag ccatttccgc tcgccgcagt
     7081 cgaacgaccg agcgtagcga gtcagtgagc gaggaagcgg aatatatcct gtatcacata
     7141 ttctgctgac gcaccggtgc agcctttttt ctcctgccac atgaagcact tcactgacac
     7201 cctcatcagt gccaacatag taagccagta tacactccgc tagcgctgat gtccggcggt
     7261 gcttttgccg ttacgcacca ccccgtcatg agattacgcg ttttagcggt gcgtacaatt
     7321 aagggattat ggtaaatcca cttactgtct gccctcgtag ccatcgagat aaaccgcagt
     7381 actccggcca cgatgcgtcc ggcgtagagg atcgagatct
//
