LOCUS       sequence-470503-        7019 bp    DNA     circular SYN 02-MAY-2025
DEFINITION  Express DGKB-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 7019)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 7019)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: DGKb-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..7019
                     /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             3498..4010
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(4098..4116)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4153..4274
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4164..4183)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4218..4237
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4519..4876
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4519..4710
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4542..4562)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4727..4862
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4891..4938
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4970..5764
                     /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(5024..5043)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5634..5653
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5828..5876
                     /note="synthetic polyadenylation signal"
     rep_origin      6033..6578
                     /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     6479..6498
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6695..6712
                     /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 cgccatgaca aaccaggaaa
     1081 aatgggccca cctcagccct tcggaatttt cccaacttca gaaatatgct gagtattcta
     1141 caaagaaatt aaaggatgtt cttgaagaat tccatggtaa tggtgtgctt gcaaagtata
     1201 atcctgaagg gaaacaagac attcttaacc aaacaataga ttttgaaggt ttcaaactat
     1261 tcatgaagac attcctggaa gccgagcttc ctgatgattt cactgcacac cttttcatgt
     1321 catttagcaa caagtttcct cattctagtc caatggtaaa aagtaagcct gctctcctat
     1381 caggcggtct gagaatgaat aaaggtgcca tcacccctcc ccggactact tctcctgcaa
     1441 atacgtgttc cccagaagta atccatctga aggacattgt ctgttacctg tctctgcttg
     1501 aaagaggaag acctgaggat aagcttgagt ttatgtttcg cctttatgac acggatggga
     1561 atggcttcct ggacagctcg gagctagaaa atatcatcag tcagatgatg catgttgcag
     1621 aataccttga gtgggatgtc actgaactta atccaatcct ccatgaaatg atggaagaaa
     1681 ttgactatga tcatgatgga accgtgtctc tggaggaatg gattcaagga ggaatgacaa
     1741 cgattccact tcttgtgctc ctgggcttag aaaataacgt gaaggatgat ggacagcacg
     1801 tgtggcgact gaagcacttt aacaaacctg cctattgcaa cctttgcctg aacatgctga
     1861 ttggcgtggg gaagcagggc ctctgctgtt ccttctgcaa gtacacagtc catgagcgct
     1921 gtgtggctcg agcacctccc tcttgcatca agacctatgt gaagtccaaa aggaacactg
     1981 atgtcatgca ccattactgg gttgaaggta actgcccaac caagtgtgat aagtgccaca
     2041 aaactgttaa atgttaccag ggcctgacag gactgcattg tgtttggtgt cagatcacac
     2101 tgcataataa atgtgcttct catctaaaac ctgaatgtga ctgtggacct ttgaaggacc
     2161 atattttacc acccacaaca atctgtccag tggtactgca gactctgccc acttcaggag
     2221 tttcagttcc tgaggaaaga caatcaacag tgaaaaagga aaagagtggt tcccagcagc
     2281 caaacaaagt gattgacaag aataaaatgc aaagagccaa ctctgttact gtagatggac
     2341 aaggcctgca ggtcactcct gtgcctggta ctcacccact tttagttttt gtgaacccca
     2401 aaagtggtgg aaaacaagga gaacgaattt acagaaaatt ccagtatcta ttaaatcctc
     2461 gtcaggttta cagtctttct ggaaatggac caatgccagg gttaaacttt ttccgtgatg
     2521 ttcctgactt cagagtgtta gcctgtggtg gagatggaac cgtgggctgg gttttggatt
     2581 gcatagaaaa ggccaatgta ggcaagcatc ctccagttgc gattctgcct cttgggactg
     2641 gcaatgatct agcaagatgc ctgcgatggg gaggaggtta cgaaggtgag aatctgatga
     2701 aaattctaaa agacattgaa aacagcacag aaatcatgtt ggacaggtgg aagtttgaag
     2761 tcatacctaa tgacaaagat gagaaaggag acccagtgcc ttacagtatc atcaataatt
     2821 acttttccat tggcgtggat gcctccattg cacacagatt ccacatcatg agagaaaaac
     2881 acccagagaa attcaacagt agaatgaaga acaaattttg gtattttgag tttggcacat
     2941 ctgaaacttt ctcagccacc tgcaagaagc tacatgaatc tgtagaaata gaatgtgatg
     3001 gagtacagat agatttaata aacatctctc tggaaggaat tgctattttg aatataccaa
     3061 gcatgcatgg aggatccaat ctttggggag agtctaagaa aagacgaagc catcgacgaa
     3121 tagagaaaaa agggtctgac aaaaggacca ccgtcacaga tgccaaagag ttgaagtttg
     3181 caagtcaaga tctcagtgac cagctgctgg aggtggtcgg cttggaagga gccatggaga
     3241 tggggcaaat atacacaggc ctgaaaagtg ctggccggcg gctggctcag tgctcctgcg
     3301 tggtcatcag gacgagcaag tctctgccaa tgcaaattga tggggagcca tggatgcaga
     3361 ccccatgcac aataaaaatt acacacaaga accaagcccc aatgctgatg ggcccgcctc
     3421 caaaaaccgg tttattctgc tccctcgtca aaaggacaag aaaccgaagc aaggaagttt
     3481 ctctcggctc gagcggcgtc ttcacactcg aagatttcgt tggggactgg cgacagacag
     3541 ccggctacaa cctggaccaa gtccttgaac agggaggtgt gtccagtttg tttcagaatc
     3601 tcggggtgtc cgtaactccg atccaaagga ttgtcctgag cggtgaaaat gggctgaaga
     3661 tcgacatcca tgtcatcatc ccgtatgaag gtctgagcgg cgaccaaatg ggccagatcg
     3721 aaaaaatttt taaggtggtg taccctgtgg atgatcatca ctttaaggtg atcctgcact
     3781 atggcacact ggtaatcgac ggggttacgc cgaacatgat cgactatttc ggacggccgt
     3841 atgaaggcat cgccgtgttc gacggcaaaa agatcactgt aacagggacc ctgtggaacg
     3901 gcaacaaaat tatcgacgag cgcctgatca accccgacgg ctccctgctg ttccgagtaa
     3961 ccatcaacgg agtgaccggc tggcggctgt gcgaacgcat tctggcgtaa ggccgcgact
     4021 ctagagtcga cctgcaggca tgcaagctga tccggctgct aacaaagccc gaaaggaagc
     4081 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cggccgcttc
     4141 gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     4201 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     4261 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga
     4321 tgtgggaggt ttttttaagc aagtaaaacc tctacaaatg tggtaaaatc gaattttaac
     4381 aaaatattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4441 tttcacaccg catacgcgga tctgcgcagc accatggcct gaaataacct ctgaaagagg
     4501 aacttggtta ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     4561 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4621 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4681 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4741 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4801 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4861 cctaggcttt tgcaaaaagc ttaattaact gttgacaatt aatcatcggc atagtatatc
     4921 ggcatagtat aatacgacaa ggtgaggaac taaacccagg aggcagatca tgattgaaca
     4981 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     5041 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     5101 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     5161 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     5221 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     5281 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     5341 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     5401 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     5461 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     5521 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     5581 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     5641 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     5701 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     5761 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     5821 tggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgaatc
     5881 gatagcgata aggatccgct catgcgatga taagctgtca aacatgagaa ttacaactta
     5941 tatcgtatgg ggctgacttc aggtgctaca tttgaagaga taaattgcac tgaaatctag
     6001 aaatatttta tctgattaat aagatgatct tcttgagatc gttttggtct gcgcgtaatc
     6061 tcttgctctg aaaacgaaaa aaccgccttg cagggcggtt tttcgaaggt tctctgagct
     6121 accaactctt tgaaccgagg taactggctt ggaggagcgc agtcaccaaa acttgtcctt
     6181 tcagtttagc cttaaccggc gcatgacttc aagactaact cctctaaatc aattaccagt
     6241 ggctgctgcc agtggtgctt ttgcatgtct ttccgggttg gactcaagac gatagttacc
     6301 ggataaggcg cagcggtcgg actgaacggg gggttcgtgc atacagtcca gcttggagcg
     6361 aactgcctac ccggaactga gtgtcaggcg tggaatgaga caaacgcggc cataacagcg
     6421 gaatgacacc ggtaaaccga aaggcaggaa caggagagcg cacgagggag ccgccagggg
     6481 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca ccactgattt gagcgtcaga
     6541 tttcgtgatg cttgtcaggg gggcggagcc tatggaaaaa cggctttgcc gcggccctct
     6601 cacttccctg ttaagtatct tcctggcatc ttccaggaaa tctccgcccc gttcgtaagc
     6661 catttccgct cgccgcagtc gaacgaccga gcgtagcgag tcagtgagcg aggaagcgga
     6721 atatatcctg tatcacatat tctgctgacg caccggtgca gccttttttc tcctgccaca
     6781 tgaagcactt cactgacacc ctcatcagtg ccaacatagt aagccagtat acactccgct
     6841 agcgctgatg tccggcggtg cttttgccgt tacgcaccac cccgtcatga gattacgcgt
     6901 tttagcggtg cgtacaatta agggattatg gtaaatccac ttactgtctg ccctcgtagc
     6961 catcgagata aaccgcagta ctccggccac gatgcgtccg gcgtagagga tcgagatct
//
