LOCUS       sequence-481954-        6232 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  Express TREM2-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 6232)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6232)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 4, 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            "Unpublished"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, TREM2-NanoLuc
FEATURES             Location/Qualifiers
     source          1..6232
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    17..98
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     42..61
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(275..787)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     promoter        complement(1537..1555)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1538..1555)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        1675..1978
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2133..2153
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        2186..2233
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2265..2924
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2331..2350)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(3096..3684)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3176..3195)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3810..3858
                     /note="synthetic polyadenylation signal"
     CDS             complement(3882..4676)
                     /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(3993..4012)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4603..4622
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5054..6232)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5063..6001
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(5086..5106)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 aagacgcgtg acgtcggcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
       61 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
      121 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
      181 aatgtggtat ggctgattat gatccgcggc cgcaatacgt cgacgttatc agctgacttc
      241 gtacgagagc ctaggtgttt ctagagtcgc ggccttacgc cagaatgcgt tcgcacagcc
      301 gccagccggt cactccgttg atggttactc ggaacagcag ggagccgtcg gggttgatca
      361 ggcgctcgtc gataattttg ttgccgttcc acagggtccc tgttacagtg atctttttgc
      421 cgtcgaacac ggcgatgcct tcatacggcc gtccgaaata gtcgatcatg ttcggcgtaa
      481 ccccgtcgat taccagtgtg ccatagtgca ggatcacctt aaagtgatga tcatccacag
      541 ggtacaccac cttaaaaatt ttttcgatct ggcccatttg gtcgccgctc agaccttcat
      601 acgggatgat gacatggatg tcgatcttca gcccattttc accgctcagg acaatccttt
      661 ggatcggagt tacggacacc ccgagattct gaaacaaact ggacacacct ccctgttcaa
      721 ggacttggtc caggttgtag ccggctgtct gtcgccagtc cccaacgaaa tcttcgagtg
      781 tgaagacgcc gctcgagccg agagaaaccg tgtctctcag ccctggcaga gtttggagct
      841 gataccctgg gtcatggcca cagtccagtt cactgggtgg atgtgtccct ggcttctgtc
      901 catgccaggc tgcagcccag agggcgctgg ctgctagaat cttgatgaga aagatgcagg
      961 ccaggaggag aaggatggaa gtgggtggga aggggatttc tccttccaag aggctcctgg
     1021 agatgctgtg ctccacatgg gcatcctcga agctctcaga ctccccgggg aaccagagat
     1081 ctccagcatc ccggtgatcc agggggtctg ccagcacctc caccaggacc ttcctgaggg
     1141 tgtcagcctc actgccatgg aggctctggc actggtagag acccgcatca tggggttgta
     1201 gattccgcag cgtaatggtg agagtgccac ccagggtatc gtctgtgatg gctgtgctcc
     1261 cattccacct cctcaggaag gacagcagcc acaagttgtg cgtgctgacc acacgctggc
     1321 atgggccctt ctctcccagc tggcggcacc aggccttgcg cctcccccag tgcttcatgg
     1381 agtcataggg gcaagacacc tgcagggact ggcccgccac gccctggaac actgtggtgt
     1441 tgtgggctcc ggacagctct gtgacaaaga gtaagatgag cagccggaga ggctccatgg
     1501 cgatcgctag ctgcaaaaag aacaagtagc ttgtattcta tagtgtcacc taaatgggcg
     1561 atctgacggt tcactaaacc agctctgctt atataggcct ccctggcgcg ccactagtta
     1621 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     1681 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     1741 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     1801 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtcc
     1861 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     1921 cttacgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     1981 gatgcggttt tggcagtaca ccaatgggcg tggatagcgg tttgactcac ggggatttcc
     2041 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     2101 tccaaaatgt cgtaataacc ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg
     2161 ggaggtcaaa tcgaattcta atactgttga caattaatca tcggcatagt atatcggcat
     2221 agtataatac gacaaggtga ggaactaaac ccaggaggca gatcatggag aaaaaaatca
     2281 ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc
     2341 agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa
     2401 agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc
     2461 tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga cggtgagctg gtgatatggg
     2521 atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct
     2581 ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt
     2641 gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtct
     2701 cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact
     2761 tcttcgcccc cgttttcact atgggcaaat attatacgca aggcgacaag gtgctgatgc
     2821 cgctggcgat tcaggttcat catgccgtct gtgatggctt ccatgtcggc agaatgctta
     2881 atgaattaca acagtactgc gatgagtggc agggcggggc gtaagcggga ctctggggtt
     2941 cgaaatgacc gaccaagcga cgcccaaccg gtatcagctc actcaaaggc ggtaatacgg
     3001 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3061 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3121 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3181 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3241 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3301 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3361 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3421 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3481 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     3541 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3601 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3661 acgcgcagaa aaaaaggatt tcaagaagat cctttgatct tttctacggg gtctgacgct
     3721 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3781 acctagatcc ttttatagtc cggaatcgtc acacaaaaaa ccaacacaca gatgtaatga
     3841 aaataaagat attttattgc ggccatcgtg atggctagag ttcagaagaa ctcgtcaaga
     3901 aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag
     3961 cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc
     4021 tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt
     4081 tccaccatga tattcggcaa gcaggcatcg ccgtgggtca cgacgagatc ctcgccgtcg
     4141 ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg
     4201 tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga
     4261 tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt
     4321 gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc
     4381 cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca
     4441 gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcctgcagt
     4501 tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac
     4561 agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat
     4621 agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatggtg
     4681 gcatcgattc ccggtcaaca tccagtacac ctccggcaca ccggcttccc caagggcgcc
     4741 acgctctccc accacaacat cctcaacaac ggctactggg tgggccgcac ggtcggctac
     4801 acggagcagg accgggtgtg tctgccggtg cccttctacc actgcttcgg catggtcatg
     4861 ggcaacttgg gcgccacctc ccacgccgcc tgcatcgtca tcccggcccc gtccttcgag
     4921 ccggcggcga cgctggaggc ggtgcagcgg gagaggtgca cgtccctgta cggcgtcccc
     4981 acgaagccga attccaccac actggactag atgcatgctc gacagtgtga tggatatctg
     5041 cagaattcgg ctttcacgac acctgaaatg gaagaaaaaa actttgaacc actgtctgag
     5101 gcttgagaat gaaccaagat ccaaactcaa aaagggcaaa ttccaaggag aattacatca
     5161 agtgccaagc tggcctaact tcagtctcca cccactcagt gtggggaaac tccatcgcat
     5221 aaaacccctc cccccaacct aaagacgacg tactccaaaa gctcgagaac taatcgaggt
     5281 gcctggacgg cgcccggtac tccgtggagt cacatgaagc gacggctgag gacggaaagg
     5341 cccttttcct ttgtgtgggt gactcacccg cccgctctcc cgagcgccgc gtcctccatt
     5401 ttgagctgcc tgcagcaggg ccgggaagcg gccatctttc cgctcacgca actggtgccg
     5461 accgggccag ccttgccgcc cagggcgggg cgatacacgg cggcgcgagg ccaggcacca
     5521 gagcaggccg gccagcttga gactaccccc gtccgattct cggtggccgc gctcgcaggc
     5581 cccgcctcgc cgaacatgtg cgctgggacg cacgggcccc gtcgccgccc gcggccccaa
     5641 aaaccgaaat accagtgtgc agatcttggc ccgcatttac aagactatct tgccagaaaa
     5701 aaagcgtcgc agcaggtcat caaaaatttt aaatggctag agacttatcg aaagcagcga
     5761 gacaggcgcg aaggtgccac cagattcgca cgcggcggcc ccagcgccca ggccaggcct
     5821 caactcaagc acgaggcgaa ggggctcctt aagcgcaagg cctcgaactc tcccacccac
     5881 ttccaacccg aagctcggga tcaagaatca cgtactgcag ccaggtggaa gtaattcaag
     5941 gcacgcaagg gccataaccc gtaaagaggc caggcccgcg ggaaccacac acggcactta
     6001 cctgtgttct ggcggcaaac ccgttgcgaa aaagaacgtt cacggcgact actgcactta
     6061 tatacggttc tcccccaccc tcgggaaaaa ggcggagcca gtacacgaca tcactttccc
     6121 agtttacccc gcgccacctt ctctaggcac cggttcaatt gccgacccct ccccccaact
     6181 tctcggggac tgtgggcgat gtgcgctctg cccactgacg ggcaccggag cc
//
