LOCUS       sequence-479844-        7668 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express NLRX1-NanoLuc 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 7668)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7668)
  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            "Unpublished"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: NLRX1-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..7668
                     /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             4011..4523
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(4611..4629)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4666..4787
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4677..4696)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4731..4750
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5032..5389
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5032..5223
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5055..5075)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5240..5375
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5404..5451
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5483..6277
                     /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(5537..5556)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6147..6166
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6341..6389
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(6628..7216)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6708..6727)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7332..7615
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
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 cgccatgagg tggggccacc
     1081 atttgcccag ggcctcttgg ggctctggtt ttagaagagc actccagcga ccagatgatc
     1141 gtatcccctt cctgatccac tggagttggc cccttcaagg ggagcgtccc tttgggcccc
     1201 ctagggcctt tatacgccac cacggaagct cggtagatag cgctccccca cccgggaggc
     1261 atggacggct gttccccagc gcctctgcaa ctgaagctat acagcggcac cgccggaacc
     1321 tggctgagtg gttcagccgg ctgcccaggg aggagcgcca gtttggccca acctttgccc
     1381 tagacacggt ccacgttgac cctgtgatcc gcgagagtac ccctgatgag ctacttcgcc
     1441 cacccgcgga gctggccctg gagcatcagc caccccaggc cgggctcccc ccactggcct
     1501 tgtctcagct ctttaacccg gatgcctgtg ggcgccgggt gcagacagtg gtgctgtatg
     1561 ggacagtggg cacaggcaag agcacgctgg tgcgcaagat ggttctggac tggtgttatg
     1621 ggcggctgcc ggccttcgag ctgctcatcc ccttctcctg tgaggacctg tcatccctgg
     1681 gccctgcccc agcctccctg tgccaacttg tggcccagcg ctacacgccc ctgaaggagg
     1741 ttctgcccct gatggctgct gctgggtccc acctcctctt tgtgctccat ggcttagagc
     1801 atctcaacct cgacttccgg ctggcaggca cgggactttg tagtgacccg gaggaaccgc
     1861 aggaaccagc tgctatcatc gtcaacctgc tgcgcaaata catgctgcct caggccagca
     1921 ttctggtgac cactcggccc tctgccattg gccgtatccc cagcaagtac gtgggccgct
     1981 atggtgagat ctgcggtttc tctgatacca acctgcagaa gctctacttc cagctccgcc
     2041 tcaaccagcc gtactgcggg tatgccgttg gcggttcagg tgtctctgcc acaccagctc
     2101 agcgtgacca cctggtgcag atgctctccc ggaacctgga ggggcaccac cagatagccg
     2161 ctgcctgctt cctgccgtcc tattgctggc tcgtttgtgc caccttgcac ttcctgcatg
     2221 cccccacgcc tgctgggcag acccttacaa gcatctatac cagcttcctg cgcctcaact
     2281 tcagcgggga aaccctggac agcactgacc cctccaattt gtccctgatg gcctatgcag
     2341 cccgaaccat gggcaagttg gcctatgagg gggtgtcctc ccgcaagacc tacttctctg
     2401 aagaggatgt ctgtggctgc ctggaggctg gcatcaggac ggaggaggag tttcagctgc
     2461 tgcacatctt ccgtcgggat gccctgaggt ttttcctggc cccatgtgtg gagccagggc
     2521 gtgcaggcac cttcgtgttc accgtgcccg ccatgcagga atacctggct gccctctaca
     2581 ttgtgctggg tttgcgcaag acgaccctgc aaaaggtggg caaggaagtg gctgagctcg
     2641 tgggccgtgt tggggaggac gtcagcctgg tactgggcat catggccaag ctgctgcctc
     2701 tgcgggctct gcctctgctc ttcaacctga tcaaggtggt tccacgagtg tttgggcgca
     2761 tggtgggtaa aagccgggag gcggtggctc aggccatggt gctggagatg tttcgagagg
     2821 aggactacta caacgatgat gttctggacc agatgggcgc cagtatcctg ggcgtggagg
     2881 gcccccggcg ccacccagat gagccccctg aggatgaagt cttcgagctc ttccccatgt
     2941 tcatgggggg gcttctctct gcccacaacc gagctgtgct agctcagctt ggctgcccca
     3001 tcaagaacct ggatgccctg gagaatgccc aggccatcaa gaagaagctg ggcaagctgg
     3061 gccggcaggt gctgccccca tcagagctcc ttgaccacct cttcttccac tatgagttcc
     3121 agaaccagcg cttctccgct gaggtgctca gctccctgcg tcagctcaac ctggcaggtg
     3181 tgcgcatgac accagtcaag tgcacagtgg tggcagctgt gctgggcagc ggaaggcatg
     3241 ccctggatga ggtgaacttg gcctcctgcc agctagatcc tgctgggctg cgcacactcc
     3301 tgcctgtctt cctgcgtgcc cggaagctgg gcttgcaact caacagcctg ggccctgagg
     3361 cctgcaagga cctccgagac ctgttgctgc atgaccagtg ccaaattacc acactgcggc
     3421 tgtccaacaa cccgctgacg gcggcaggtg ttgccgtgct aatggagggg ctggcaggaa
     3481 acacctcagt gacgcacctg tccctgctgc acacgggcct tggggacgaa ggcctggagc
     3541 tgctggctgc ccagctggac cgcaaccggc agctgcagga gctgaacgtg gcgtacaacg
     3601 gtgctggtga cacagcggcc ctggccctgg ccagagctgc ccgggagcac ccttccctgg
     3661 aactgctaca cctctacttc aatgagctga gctcagaggg ccgccaggtc ttgcgagact
     3721 tggggggtgc tgctgaaggt ggtgcccggg tggtggtgtc actgacagag gggacggcgg
     3781 tgtcagaata ctggtcagtg atcctcagtg aagtccagcg gaacctcaat agctgggatc
     3841 gggcccgggt tcagcgacac cttgagctcc tactgcggga tctggaagat agccggggtg
     3901 ccacccttaa tccttggcgc aaggcccagc tgctgcgagt ggagggcgag gtcagggccc
     3961 tcctggagca gctgggaagc tctggaagcg tttctctcgg ctcgagcggc gtcttcacac
     4021 tcgaagattt cgttggggac tggcgacaga cagccggcta caacctggac caagtccttg
     4081 aacagggagg tgtgtccagt ttgtttcaga atctcggggt gtccgtaact ccgatccaaa
     4141 ggattgtcct gagcggtgaa aatgggctga agatcgacat ccatgtcatc atcccgtatg
     4201 aaggtctgag cggcgaccaa atgggccaga tcgaaaaaat ttttaaggtg gtgtaccctg
     4261 tggatgatca tcactttaag gtgatcctgc actatggcac actggtaatc gacggggtta
     4321 cgccgaacat gatcgactat ttcggacggc cgtatgaagg catcgccgtg ttcgacggca
     4381 aaaagatcac tgtaacaggg accctgtgga acggcaacaa aattatcgac gagcgcctga
     4441 tcaaccccga cggctccctg ctgttccgag taaccatcaa cggagtgacc ggctggcggc
     4501 tgtgcgaacg cattctggcg taaggccgcg actctagagt cgacctgcag gcatgcaagc
     4561 tgatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     4621 ataactagca taaccccttg gggcggccgc ttcgagcaga catgataaga tacattgatg
     4681 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     4741 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     4801 gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttta agcaagtaaa
     4861 acctctacaa atgtggtaaa atcgaatttt aacaaaatat taacgcttac aatttcctga
     4921 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgc ggatctgcgc
     4981 agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct tctgaggcgg
     5041 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     5101 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     5161 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     5221 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     5281 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     5341 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttaatta
     5401 actgttgaca attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg
     5461 aactaaaccc aggaggcaga tcatgattga acaagatgga ttgcacgcag gttctccggc
     5521 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     5581 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     5641 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     5701 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     5761 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     5821 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     5881 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     5941 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     6001 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     6061 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     6121 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     6181 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     6241 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     6301 accgaccaag cgacgcccaa cctgccatca cgatggccgc aataaaatat ctttattttc
     6361 attacatctg tgtgttggtt ttttgtgtga atcgatagcg ataaggatcc tctttgcgct
     6421 tgcgttttcc cttgtccaga tagcccagta gctgacattc atccggggtc agcaccgttt
     6481 ctgcggactg gctttctacc cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6541 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6601 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6661 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6721 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6781 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6841 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6901 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6961 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7021 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     7081 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7141 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7201 aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7261 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7321 ccttttatag tccggaaata caggaacgca cgctggatgg cccttcgctg ggatggtgaa
     7381 accatgaaaa atggcagctt cagtggatta agtgggggta atgtggcctg taccctctgg
     7441 ttgcataggt attcatacgg ttaaaattta tcaggcgcga ttgcggcagt ttttcgggtg
     7501 gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc tgaacgcgtt ttagcggtgc
     7561 gtacaattaa gggattatgg taaatccact tactgtctgc cctcgtagcc atcgagataa
     7621 accgcagtac tccggccacg atgcgtccgg cgtagaggat cgagatct
//
