LOCUS       sequence-472433-        7802 bp    DNA     circular SYN 13-MAY-2025
DEFINITION  Express DGKI-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 7802)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 7802)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: DGKi-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..7802
                     /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             4281..4793
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(4881..4899)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4936..5057
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4947..4966)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5001..5020
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5302..5659
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5302..5493
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5325..5345)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5510..5645
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5674..5721
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5753..6547
                     /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(5807..5826)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6417..6436
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6611..6659
                     /note="synthetic polyadenylation signal"
     rep_origin      6816..7361
                     /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     7262..7281
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7478..7495
                     /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 cgccatggat gctgctggaa
     1081 gaggttgtca tttactacct ctaccagcgg ctagaggacc tgctagggct cctgcagctg
     1141 cagctgctgc agcagcaagt cctccaggtc catgtagtgg agctgcatgt gctccaagtg
     1201 cagcagctgg agcaggtgct atgaatccta gttcatcggc gggagaggag aaaggggcga
     1261 cgggcggcag cagcagcagc ggaagcggtg ctggtagttg ttgtctagga gcagaaggtg
     1321 gtgcggaccc gcggggcgca gggtcagccg ctgcagcagg agctgctgca ctggacgaac
     1381 ctgctgcagc aggtcagaag gagaaggacg aagcgctgga ggagaagctg aggaacttaa
     1441 ctttccggaa gcaggtctcg tacaggaaag caatctcccg ggcaggcctc cagcatctgg
     1501 ctcctgcaca tcccctcagc cttcctgtgg caaatggtcc agccaaggag cccagagcga
     1561 ctttggactg gagtgagaat gccgtgaatg gagaacacct gtggctggag accaacgtct
     1621 cgggagacct ctgctacctt ggagaggaga actgccaagt cagatttgca aaatcagctc
     1681 tcaggaggaa gtgtgcagtc tgtaaaatcg tcgtccacac cgcctgcatt gagcagctag
     1741 aaaagattaa tttcagatgt aaaccaacat ttcgagaagg aggctcaagg tcaccaagag
     1801 aaaattttgt acgtcatcac tgggtgcaca ggcgtcggca ggaggggaaa tgtaagcagt
     1861 gtggtaaggg cttccagcaa aagttctcct tccacagtaa agagattgtg gctatcagct
     1921 gttcctggtg caagcaggcg tttcacaata aggtgacctg cttcatgctg catcacattg
     1981 aagaaccctg ctccctgggg gctcatgctg ctgttattgt cccgcccact tggatcatta
     2041 aggtgaagaa acctcagaac tccctgaagg cttcaaatcg gaagaagaag agaacaagct
     2101 ttaaaagaaa agccagtaaa agagggatgg aacaggaaaa caaaggtcgt ccttttgtga
     2161 taaaacccat ctcttctcct ctcatgaaac ccttgcttgt atttgtgaat cccaagagtg
     2221 gaggcaacca gggaaccaaa gtcctgcaga tgttcatgtg gtacctgaat ccacggcaag
     2281 tctttgatct ttctcaggaa gggccaaaag atgcgcttga attgtatagg aaagtaccaa
     2341 atctgcgaat tctggcctgt ggtggggatg gaacggtggg ctggatcctt tccatcctgg
     2401 atgaactgca gctgaaccct cagcctcctg tgggggtcct tcctctgggg actgggaatg
     2461 acctggctcg aactctcaac tggggagggg gctacactga tgaacctgtt tctaaaattc
     2521 tttgtcaagt tgaagatggt acagtagtac aactagatcg ttggaatctc catgtggaaa
     2581 gaaaccccga cttgcctcca gaagaacttg aagatggcgt atgtaagctc cctctgaatg
     2641 ttttcaataa ctacttcagc cttggatttg atgcccatgt cacactggag ttccatgaat
     2701 ccagagaagc aaatccagag aaattcaaca gtcgttttcg aaataaaatg ttctatgcag
     2761 gggcagcttt ttctgacttc ctacagagaa gttctagaga tctatccaaa catgttaaag
     2821 ttgtttgtga tggaacagat ctcaccccaa agattcagga actgaagttc cagtgtatag
     2881 tatttttaaa tatacccaga tattgtgctg gcacaatgcc ctggggaaac ccaggtgatc
     2941 accatgattt cgaacctcag cgtcatgatg atggttatat tgaagtcatt ggatttacca
     3001 tggcctcttt ggcagccctg caagttgggg gccatggaga gaggctacac cagtgtcgag
     3061 aagtcatgct tctaacttac aaatccatcc ccatgcaagt ggatggggag ccctgtaggt
     3121 tggccccagc tatgattcgg atctccctga ggaatcaggc caacatggta cagaagagca
     3181 agaggagaac atccatgcct ttactcaatg atccccagtc tgtcccagat cgtctgagga
     3241 tccgggtgaa caaaatcagt ttacaagact atgaaggatt ccactatgac aaggagaaac
     3301 tccgagaagc ttctatttca gactggttaa gaaccattgc tggggaacta gtgcagtcat
     3361 ttggagcgat acctctgggt attctagttg tgcgtggaga ctgtgatttg gagacttgcc
     3421 gtatgtacat agaccgccta caggaggacc tacagtcagt ttcttctggc tcccagagag
     3481 ttcattacca ggaccatgaa acctccttcc ccagggctct ctcagcacag aggctctctc
     3541 ctcggtggtg cttcctagat gacagatctc aggaacattt gcactttgtg atggagattt
     3601 cccaagatga gatttttatt ctggacccag atatggtggt gtcacagccg gcggggacac
     3661 ctccgggcat gcctgacctg gtggtggaac aagcctcggg gatctcagac tggtggaatc
     3721 ctgccctgcg gaaacgcatg ctgagtgaca gtgggctggg gatgatagct ccctattatg
     3781 aggactcaga tctgaaagat ctcagccact cccgcgtgct acagtcacca gtctcttcag
     3841 aagatcatgc aattttgcag gcagtaatag ctggtgatct tatgaagcta atagaaagct
     3901 ataaaaatgg aggcagtctg ctaattcagg gaccagacca ctgttcactc cttcactacg
     3961 cagctaaaac cggcaacggg gagattgtga aatatatcct tgaccacgga ccttccgagt
     4021 tattggatat ggcagacagt gaaacgggtg agactgcact gcacaaggct gcctgccagc
     4081 ggaaccgggc tgtgtgccag cttctggtgg atgcaggagc atctctgaga aagacggact
     4141 ccaagggtaa gacacctcaa gaaagagcac agcaggctgg ggacccagac ttggctgctt
     4201 acctagaaag ccgtcagaac tataaggtca ttggccatga ggacctggaa actgctgttg
     4261 tttctctcgg ctcgagcggc gtcttcacac tcgaagattt cgttggggac tggcgacaga
     4321 cagccggcta caacctggac caagtccttg aacagggagg tgtgtccagt ttgtttcaga
     4381 atctcggggt gtccgtaact ccgatccaaa ggattgtcct gagcggtgaa aatgggctga
     4441 agatcgacat ccatgtcatc atcccgtatg aaggtctgag cggcgaccaa atgggccaga
     4501 tcgaaaaaat ttttaaggtg gtgtaccctg tggatgatca tcactttaag gtgatcctgc
     4561 actatggcac actggtaatc gacggggtta cgccgaacat gatcgactat ttcggacggc
     4621 cgtatgaagg catcgccgtg ttcgacggca aaaagatcac tgtaacaggg accctgtgga
     4681 acggcaacaa aattatcgac gagcgcctga tcaaccccga cggctccctg ctgttccgag
     4741 taaccatcaa cggagtgacc ggctggcggc tgtgcgaacg cattctggcg taaggccgcg
     4801 actctagagt cgacctgcag gcatgcaagc tgatccggct gctaacaaag cccgaaagga
     4861 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcggccgc
     4921 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     4981 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     5041 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     5101 agatgtggga ggttttttta agcaagtaaa acctctacaa atgtggtaaa atcgaatttt
     5161 aacaaaatat taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg
     5221 gtatttcaca ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa cctctgaaag
     5281 aggaacttgg ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     5341 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     5401 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     5461 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     5521 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     5581 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     5641 aggcctaggc ttttgcaaaa agcttaatta actgttgaca attaatcatc ggcatagtat
     5701 atcggcatag tataatacga caaggtgagg aactaaaccc aggaggcaga tcatgattga
     5761 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     5821 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     5881 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     5941 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     6001 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     6061 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     6121 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     6181 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     6241 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     6301 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     6361 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     6421 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     6481 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     6541 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     6601 cgatggccgc aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga
     6661 atcgatagcg ataaggatcc gctcatgcga tgataagctg tcaaacatga gaattacaac
     6721 ttatatcgta tggggctgac ttcaggtgct acatttgaag agataaattg cactgaaatc
     6781 tagaaatatt ttatctgatt aataagatga tcttcttgag atcgttttgg tctgcgcgta
     6841 atctcttgct ctgaaaacga aaaaaccgcc ttgcagggcg gtttttcgaa ggttctctga
     6901 gctaccaact ctttgaaccg aggtaactgg cttggaggag cgcagtcacc aaaacttgtc
     6961 ctttcagttt agccttaacc ggcgcatgac ttcaagacta actcctctaa atcaattacc
     7021 agtggctgct gccagtggtg cttttgcatg tctttccggg ttggactcaa gacgatagtt
     7081 accggataag gcgcagcggt cggactgaac ggggggttcg tgcatacagt ccagcttgga
     7141 gcgaactgcc tacccggaac tgagtgtcag gcgtggaatg agacaaacgc ggccataaca
     7201 gcggaatgac accggtaaac cgaaaggcag gaacaggaga gcgcacgagg gagccgccag
     7261 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccaccactga tttgagcgtc
     7321 agatttcgtg atgcttgtca ggggggcgga gcctatggaa aaacggcttt gccgcggccc
     7381 tctcacttcc ctgttaagta tcttcctggc atcttccagg aaatctccgc cccgttcgta
     7441 agccatttcc gctcgccgca gtcgaacgac cgagcgtagc gagtcagtga gcgaggaagc
     7501 ggaatatatc ctgtatcaca tattctgctg acgcaccggt gcagcctttt ttctcctgcc
     7561 acatgaagca cttcactgac accctcatca gtgccaacat agtaagccag tatacactcc
     7621 gctagcgctg atgtccggcg gtgcttttgc cgttacgcac caccccgtca tgagattacg
     7681 cgttttagcg gtgcgtacaa ttaagggatt atggtaaatc cacttactgt ctgccctcgt
     7741 agccatcgag ataaaccgca gtactccggc cacgatgcgt ccggcgtaga ggatcgagat
     7801 ct
//
