LOCUS       sequence-471910-        6508 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express NanoLuc(R)-IRAK2 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 6508)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6508)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoLuc-IRAK2 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6508
                     /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"
     primer_bind     complement(3587..3605)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3642..3763
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3653..3672)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3707..3726
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4008..4365
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4008..4199
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4031..4051)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4216..4351
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4380..4427
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4459..5253
                     /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(4513..4532)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5123..5142
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5317..5365
                     /note="synthetic polyadenylation signal"
     rep_origin      5522..6067
                     /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     5968..5987
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6184..6201
                     /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 ggcctgctac atctatcagc
     1621 tgccaagctg ggtgctggac gacctgtgcc ggaacatgga cgccctgtcc gagtgggatt
     1681 ggatggagtt cgcctcttac gtgatcaccg acctgacaca gctgcggaag atcaagtcta
     1741 tggagagagt gcagggcgtg agcatcaccc gggagctgct gtggtggtgg ggcatgagac
     1801 aggccacagt gcagcagctg gtggatctgc tgtgcaggct ggagctgtat cgcgccgccc
     1861 agatcatcct gaattggaag cctgccccag agatcaggtg ccccatccct gcctttcccg
     1921 acagcgtgaa gcctgagaag ccactggcag catccgtgag gaaggcagag gatgagcagg
     1981 aggagggaca gcctgtgcgc atggccacct tcccaggacc aggcagctcc ccagcacggg
     2041 cccaccagcc tgcctttctg cagccacctg aggaggacgc accacactct ctgcgcagcg
     2101 atctgcccac ctctagcgac tctaaggatt tcagcacatc catccctaag caggagaagc
     2161 tgctgagcct ggccggcgac tccctgtttt ggtctgaggc cgatgtggtg caggccaccg
     2221 acgatttcaa ccagaatagg aagatctccc agggcacatt tgccgacgtg tacaggggac
     2281 acagacacgg caagcccttc gtgtttaaga agctgaggga gaccgcatgc tcctctcctg
     2341 gaagcatcga gaggttcttt caggccgagc tgcagatctg cctgcgctgc tgtcacccaa
     2401 acgtgctgcc cgtgctggga ttctgtgcag caagacagtt ccactctttt atctacccct
     2461 atatggccaa tggcagcctg caggacagac tgcagggaca gggaggatcc gatcctctgc
     2521 catggcccca gagggtgtcc atctgctctg gcctgctgtg cgccgtggag tatctgcacg
     2581 gcctggagat catccacagc aacgtgaaga gcagcaatgt gctgctggat cagaacctga
     2641 cccctaagct ggcacaccca atggcacacc tgtgccctgt gaataagcgg agcaagtaca
     2701 ccatgatgaa gacacacctg ctgagaacct ccgccgccta tctgccagag gacttcatca
     2761 gggtgggcca gctgacaaag cgcgtggata tcttttcctg cggcatcgtg ctggccgagg
     2821 tgctgacagg catccctgcc atggacaaca atcgctcccc agtgtacctg aaggacctgc
     2881 tgctgtctga tatcccatct agcaccgcaa gcctgtgctc caggaagaca ggagtggaga
     2941 acgtgatggc caaggagatc tgtcagaagt atctggagaa gggagcaggc agactgccag
     3001 aggattgcgc agaggccctg gcaaccgcag catgcctgtg cctgcggaga aggaatacaa
     3061 gcctgcagga ggtgtgcgga tccgtggcag cagtggagga gcgcctgcgg ggcagagaga
     3121 ccctgctgcc atggtctggc ctgagcgagg gcacaggctc ctctagcaac acccccgagg
     3181 agacagacga tgtggacaat tcctctctgg atgccagctc ctctatgagc gtggcacctt
     3241 gggcaggagc agcaaccccc ctgctgccta cagagaacgg cgagggcagg ctgcgcgtga
     3301 tcgtgggaag ggaggcagac agctcctctg aggcatgcgt gggactggag ccaccacagg
     3361 atgtgaccga gacatcctgg cagatcgaga tcaacgaggc caagagaaag ctgatggaga
     3421 atatcctgct gtacaaggag gagaaggtgg actctatcga gctgtttggc ccagtttaaa
     3481 cgaattcggg ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat gcaagctgat
     3541 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     3601 ctagcataac cccttggggc ggccgcttcg agcagacatg ataagataca ttgatgagtt
     3661 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3721 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3781 tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca agtaaaacct
     3841 ctacaaatgt ggtaaaatcg aattttaaca aaatattaac gcttacaatt tcctgatgcg
     3901 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgcggat ctgcgcagca
     3961 ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg aggcggaaag
     4021 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4081 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4141 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4201 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4261 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     4321 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct taattaactg
     4381 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact
     4441 aaacccagga ggcagatcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4501 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4561 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4621 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     4681 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4741 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4801 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4861 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4921 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4981 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     5041 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     5101 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     5161 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     5221 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     5281 accaagcgac gcccaacctg ccatcacgat ggccgcaata aaatatcttt attttcatta
     5341 catctgtgtg ttggtttttt gtgtgaatcg atagcgataa ggatccgctc atgcgatgat
     5401 aagctgtcaa acatgagaat tacaacttat atcgtatggg gctgacttca ggtgctacat
     5461 ttgaagagat aaattgcact gaaatctaga aatattttat ctgattaata agatgatctt
     5521 cttgagatcg ttttggtctg cgcgtaatct cttgctctga aaacgaaaaa accgccttgc
     5581 agggcggttt ttcgaaggtt ctctgagcta ccaactcttt gaaccgaggt aactggcttg
     5641 gaggagcgca gtcaccaaaa cttgtccttt cagtttagcc ttaaccggcg catgacttca
     5701 agactaactc ctctaaatca attaccagtg gctgctgcca gtggtgcttt tgcatgtctt
     5761 tccgggttgg actcaagacg atagttaccg gataaggcgc agcggtcgga ctgaacgggg
     5821 ggttcgtgca tacagtccag cttggagcga actgcctacc cggaactgag tgtcaggcgt
     5881 ggaatgagac aaacgcggcc ataacagcgg aatgacaccg gtaaaccgaa aggcaggaac
     5941 aggagagcgc acgagggagc cgccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6001 gtttcgccac cactgatttg agcgtcagat ttcgtgatgc ttgtcagggg ggcggagcct
     6061 atggaaaaac ggctttgccg cggccctctc acttccctgt taagtatctt cctggcatct
     6121 tccaggaaat ctccgccccg ttcgtaagcc atttccgctc gccgcagtcg aacgaccgag
     6181 cgtagcgagt cagtgagcga ggaagcggaa tatatcctgt atcacatatt ctgctgacgc
     6241 accggtgcag ccttttttct cctgccacat gaagcacttc actgacaccc tcatcagtgc
     6301 caacatagta agccagtata cactccgcta gcgctgatgt ccggcggtgc ttttgccgtt
     6361 acgcaccacc ccgtcatgag attacgcgtt ttagcggtgc gtacaattaa gggattatgg
     6421 taaatccact tactgtctgc cctcgtagcc atcgagataa accgcagtac tccggccacg
     6481 atgcgtccgg cgtagaggat cgagatct
//
