LOCUS       sequence-473355-        6980 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express DGKG-NanoLuc(R) 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 6980)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6980)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: DGKg-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6980
                     /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             3459..3971
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(4059..4077)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4114..4235
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4125..4144)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4179..4198
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4480..4837
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4480..4671
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4503..4523)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4688..4823
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4852..4899
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4931..5725
                     /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(4985..5004)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5595..5614
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5789..5837
                     /note="synthetic polyadenylation signal"
     rep_origin      5994..6539
                     /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     6440..6459
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6656..6673
                     /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 cgccatgggt gaagaacggt
     1081 gggtctccct cactccagaa gaatttgacc aactccagaa atattcagaa tattcctcca
     1141 agaagataaa agatgccttg actgaattta atgagggtgg gagcctcaaa caatatgacc
     1201 cacatgagcc gattagctat gatgtcttca agctgttcat gagggcgtac ctggaggtgg
     1261 accttcccca gccactgagc actcacctct tcctggcctt cagccagaag cccagacacg
     1321 agacctctga ccacccgacg gagggagcca gcaacagtga ggccaacagc gcagatacta
     1381 atatacagaa tgcagataat gccaccaaag cagacgaggc ctgtgcccct gatactgaat
     1441 caaatatggc tgagaagcaa gcaccagctg aagaccaagt ggctgcgacc cccctggaac
     1501 cccccgtccc tcggtcttca agctcggaat ccccagtggt atacctgaag gatgttgtgt
     1561 gctacctgtc cctgctggag acggggaggc ctcaggataa gctggagttc atgtttcgcc
     1621 tctatgattc agatgagaac ggtctcctgg accaagcgga gatggattgc attgtcaacc
     1681 aaatgctgca tattgcccag tacctggagt gggatcccac agagctgagg cctatattga
     1741 aggagatgct gcaagggatg gactacgacc gggacggctt tgtgtctcta caggaatggg
     1801 tccatggagg gatgaccacc atcccattgc tggtcctcct ggggatggat gactctggct
     1861 ccaaggggga tgggcggcac gcctggacca tgaagcactt caagaaacca acctactgca
     1921 acttctgcca tatcatgctc atgggcgtcc gcaagcaagg cctgtgctgc acttactgta
     1981 aatacactgt ccacgaacgc tgtgtgtcca gaaacattcc tggttgtgtc aaaacgtact
     2041 caaaagccaa aaggagtggt gaggtgatgc agcacgcatg ggtggaaggg aactcctccg
     2101 tcaagtgtga ccggtgccac aaaagtatca agtgctacca gagtgtcacc gcgcggcact
     2161 gcgtgtggtg ccggatgacg tttcaccgca aatgtgaatt atcaacgttg tgtgacggtg
     2221 gggaactcag agaccacatc ttactgccca cctccatatg ccccatcacc cgggacaggc
     2281 caggtgagaa gtctgatggc tgcgtgtccg ccaagggcga acttgtcatg cagtataaga
     2341 tcatccccac cccgggtacc caccccctgc tggtcttggt gaaccccaag agtggaggga
     2401 gacaaggaga aagaattctt cggaaattcc actatctgct caaccccaaa caagttttca
     2461 acctggacaa tggggggcct actccagggt tgaacttttt ccgtgatact ccagacttcc
     2521 gtgttttggc ctgtggtgga gatgggacag ttggctggat tttggattgc attgataagg
     2581 ccaactttgc aaagcatcca ccagtggctg tcctgcctct tggaacagga aatgaccttg
     2641 cccgttgtct ccgctgggga ggaggttatg aagggggcag cttgacaaaa atcctgaaag
     2701 acattgagca gagccccttg gtgatgctgg accgctggca tctggaagtc atccccagag
     2761 aggaagtgga aaacggggac caggtcccat acagcatcat gaacaactat ttctccattg
     2821 gtgtggacgc ttccattgca cacagattcc atgtgatgag agagaaacat cctgaaaaat
     2881 tcaacagcag gatgaagaac aagctgtggt actttgaatt tggcacctcg gagacttttg
     2941 cagcgacctg caagaaactc cacgaccaca ttgagttgga gtgtgatggg gttggggtgg
     3001 acctgagcaa catcttcctg gaaggcattg ccattctcaa cattcccagc atgtacggag
     3061 gcaccaatct ctggggagaa aacaagaaga accgggctgt gatccgggaa agcaggaagg
     3121 gtgtcactga ccccaaagaa ctgaaattct gcgttcaaga cctcagtgac cagctccttg
     3181 aagtggtggg gctagaagga gccatggaga tggggcagat ctacaccggc ctgaagagtg
     3241 caggcaggag gctggcccag tgcgcctctg tcaccatcag gacaaacaag ctgctgccaa
     3301 tgcaagtgga tggagaaccc tggatgcagc catgttgcac gattaaaatt actcacaaga
     3361 accaagcgcc catgatgatg gggcctcccc agaagagcag cttcttctcg ttgagaagga
     3421 agagccgttc aaaagacgtt tctctcggct cgagcggcgt cttcacactc gaagatttcg
     3481 ttggggactg gcgacagaca gccggctaca acctggacca agtccttgaa cagggaggtg
     3541 tgtccagttt gtttcagaat ctcggggtgt ccgtaactcc gatccaaagg attgtcctga
     3601 gcggtgaaaa tgggctgaag atcgacatcc atgtcatcat cccgtatgaa ggtctgagcg
     3661 gcgaccaaat gggccagatc gaaaaaattt ttaaggtggt gtaccctgtg gatgatcatc
     3721 actttaaggt gatcctgcac tatggcacac tggtaatcga cggggttacg ccgaacatga
     3781 tcgactattt cggacggccg tatgaaggca tcgccgtgtt cgacggcaaa aagatcactg
     3841 taacagggac cctgtggaac ggcaacaaaa ttatcgacga gcgcctgatc aaccccgacg
     3901 gctccctgct gttccgagta accatcaacg gagtgaccgg ctggcggctg tgcgaacgca
     3961 ttctggcgta aggccgcgac tctagagtcg acctgcaggc atgcaagctg atccggctgc
     4021 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
     4081 accccttggg gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa
     4141 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     4201 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     4261 tgtttcaggt tcagggggag atgtgggagg tttttttaag caagtaaaac ctctacaaat
     4321 gtggtaaaat cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc
     4381 tccttacgca tctgtgcggt atttcacacc gcatacgcgg atctgcgcag caccatggcc
     4441 tgaaataacc tctgaaagag gaacttggtt aggtaccttc tgaggcggaa agaaccagct
     4501 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4561 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4621 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4681 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4741 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     4801 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag cttaattaac tgttgacaat
     4861 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaacccag
     4921 gaggcagatc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4981 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     5041 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     5101 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     5161 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     5221 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     5281 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     5341 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     5401 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     5461 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     5521 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     5581 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     5641 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     5701 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     5761 acgcccaacc tgccatcacg atggccgcaa taaaatatct ttattttcat tacatctgtg
     5821 tgttggtttt ttgtgtgaat cgatagcgat aaggatccgc tcatgcgatg ataagctgtc
     5881 aaacatgaga attacaactt atatcgtatg gggctgactt caggtgctac atttgaagag
     5941 ataaattgca ctgaaatcta gaaatatttt atctgattaa taagatgatc ttcttgagat
     6001 cgttttggtc tgcgcgtaat ctcttgctct gaaaacgaaa aaaccgcctt gcagggcggt
     6061 ttttcgaagg ttctctgagc taccaactct ttgaaccgag gtaactggct tggaggagcg
     6121 cagtcaccaa aacttgtcct ttcagtttag ccttaaccgg cgcatgactt caagactaac
     6181 tcctctaaat caattaccag tggctgctgc cagtggtgct tttgcatgtc tttccgggtt
     6241 ggactcaaga cgatagttac cggataaggc gcagcggtcg gactgaacgg ggggttcgtg
     6301 catacagtcc agcttggagc gaactgccta cccggaactg agtgtcaggc gtggaatgag
     6361 acaaacgcgg ccataacagc ggaatgacac cggtaaaccg aaaggcagga acaggagagc
     6421 gcacgaggga gccgccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6481 accactgatt tgagcgtcag atttcgtgat gcttgtcagg ggggcggagc ctatggaaaa
     6541 acggctttgc cgcggccctc tcacttccct gttaagtatc ttcctggcat cttccaggaa
     6601 atctccgccc cgttcgtaag ccatttccgc tcgccgcagt cgaacgaccg agcgtagcga
     6661 gtcagtgagc gaggaagcgg aatatatcct gtatcacata ttctgctgac gcaccggtgc
     6721 agcctttttt ctcctgccac atgaagcact tcactgacac cctcatcagt gccaacatag
     6781 taagccagta tacactccgc tagcgctgat gtccggcggt gcttttgccg ttacgcacca
     6841 ccccgtcatg agattacgcg ttttagcggt gcgtacaatt aagggattat ggtaaatcca
     6901 cttactgtct gccctcgtag ccatcgagat aaaccgcagt actccggcca cgatgcgtcc
     6961 ggcgtagagg atcgagatct
//
