LOCUS       sequence-479784-        5445 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express TREM1-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 5445)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5445)
  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            "Unpublished"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: NanoBRET Assay Vector, TREM1-NanoLuc
FEATURES             Location/Qualifiers
     source          1..5445
                     /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             1788..2300
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2388..2406)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2443..2564
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2454..2473)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2508..2527
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2809..3166
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2809..3000
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2832..2852)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3017..3152
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3181..3228
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3260..4054
                     /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(3314..3333)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3924..3943
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4118..4166
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4405..4993)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4485..4504)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5109..5392
                     /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 aagaccaggc
     1081 tctgggggct gctgtggatg ctctttgtct cagaactccg agctgcaact aaattaactg
     1141 aggaaaagta tgaactgaaa gaggggcaga ccctggatgt gaaatgtgac tacacgctag
     1201 agaagtttgc cagcagccag aaagcttggc agataataag ggacggagag atgcccaaga
     1261 ccctggcatg cacagagagg ccttcaaaga attcccatcc agtccaagtg gggaggatca
     1321 tactagaaga ctaccatgat catggtttac tgcgcgtccg aatggtcaac cttcaagtgg
     1381 aagattctgg actgtatcag tgtgtgatct accagcctcc caaggagcct cacatgctgt
     1441 tcgatcgcat ccgcttggtg gtgaccaagg gtttttcagg gacccctggc tccaatgaga
     1501 attctaccca gaatgtgtat aagattcctc ctaccaccac taaggccttg tgcccactct
     1561 ataccagccc cagaactgtg acccaagctc cacccaagtc aactgccgat gtctccactc
     1621 ctgactctga aatcaacctt acaaatgtga cagatatcat cagggttccg gtgttcaaca
     1681 ttgtcattct cctggctggt ggattcctga gtaagagcct ggtcttctct gtcctgtttg
     1741 ctgtcacgct gaggtcattt gtacccgttt ctctcggctc gagcggcgtc ttcacactcg
     1801 aagatttcgt tggggactgg cgacagacag ccggctacaa cctggaccaa gtccttgaac
     1861 agggaggtgt gtccagtttg tttcagaatc tcggggtgtc cgtaactccg atccaaagga
     1921 ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca tgtcatcatc ccgtatgaag
     1981 gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt taaggtggtg taccctgtgg
     2041 atgatcatca ctttaaggtg atcctgcact atggcacact ggtaatcgac ggggttacgc
     2101 cgaacatgat cgactatttc ggacggccgt atgaaggcat cgccgtgttc gacggcaaaa
     2161 agatcactgt aacagggacc ctgtggaacg gcaacaaaat tatcgacgag cgcctgatca
     2221 accccgacgg ctccctgctg ttccgagtaa ccatcaacgg agtgaccggc tggcggctgt
     2281 gcgaacgcat tctggcgtaa ggccgcgact ctagagtcga cctgcaggca tgcaagctga
     2341 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
     2401 actagcataa ccccttgggg cggccgcttc gagcagacat gataagatac attgatgagt
     2461 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     2521 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     2581 ttcattttat gtttcaggtt cagggggaga tgtgggaggt ttttttaagc aagtaaaacc
     2641 tctacaaatg tggtaaaatc gaattttaac aaaatattaa cgcttacaat ttcctgatgc
     2701 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgcgga tctgcgcagc
     2761 accatggcct gaaataacct ctgaaagagg aacttggtta ggtaccttct gaggcggaaa
     2821 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     2881 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     2941 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3001 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3061 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     3121 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttaattaact
     3181 gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac
     3241 taaacccagg aggcagatca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3301 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3361 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     3421 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     3481 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     3541 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     3601 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     3661 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     3721 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     3781 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     3841 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     3901 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     3961 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4021 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     4081 gaccaagcga cgcccaacct gccatcacga tggccgcaat aaaatatctt tattttcatt
     4141 acatctgtgt gttggttttt tgtgtgaatc gatagcgata aggatcctct ttgcgcttgc
     4201 gttttccctt gtccagatag cccagtagct gacattcatc cggggtcagc accgtttctg
     4261 cggactggct ttctacccgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4321 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4381 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4441 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4501 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4561 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4621 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4681 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4741 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4801 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     4861 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4921 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4981 aaaaggattt caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5041 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5101 tttatagtcc ggaaatacag gaacgcacgc tggatggccc ttcgctggga tggtgaaacc
     5161 atgaaaaatg gcagcttcag tggattaagt gggggtaatg tggcctgtac cctctggttg
     5221 cataggtatt catacggtta aaatttatca ggcgcgattg cggcagtttt tcgggtggtt
     5281 tgttgccatt tttacctgtc tgctgccgtg atcgcgctga acgcgtttta gcggtgcgta
     5341 caattaaggg attatggtaa atccacttac tgtctgccct cgtagccatc gagataaacc
     5401 gcagtactcc ggccacgatg cgtccggcgt agaggatcga gatct
//
