LOCUS       sequence-479777-        5491 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express NanoLuc-TREM2 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 5491)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5491)
  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            "Unpublished"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: NanoLuc-TREM2 Vector
FEATURES             Location/Qualifiers
     source          1..5491
                     /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             1154..1663
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2434..2452)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2489..2610
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2500..2519)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2554..2573
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2855..3212
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2855..3046
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2878..2898)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3063..3198
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3227..3274
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3306..4100
                     /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(3360..3379)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3970..3989
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4164..4212
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4451..5039)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4531..4550)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5155..5438
                     /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 ggctagcaaa gccaccatga actccttctc
     1081 cacaagcgcc ttcggtccag ttgccttctc cctgggcctg ctcctggtgt tgcctgctgc
     1141 cttccctgcc ccagtcttca cactcgaaga tttcgttggg gactggcgac agacagccgg
     1201 ctacaacctg gaccaagtcc ttgaacaggg aggtgtgtcc agtttgtttc agaatctcgg
     1261 ggtgtccgta actccgatcc aaaggattgt cctgagcggt gaaaatgggc tgaagatcga
     1321 catccatgtc atcatcccgt atgaaggtct gagcggcgac caaatgggcc agatcgaaaa
     1381 aatttttaag gtggtgtacc ctgtggatga tcatcacttt aaggtgatcc tgcactatgg
     1441 cacactggta atcgacgggg ttacgccgaa catgatcgac tatttcggac ggccgtatga
     1501 aggcatcgcc gtgttcgacg gcaaaaagat cactgtaaca gggaccctgt ggaacggcaa
     1561 caaaattatc gacgagcgcc tgatcaaccc cgacggctcc ctgctgttcc gagtaaccat
     1621 caacggagtg accggctggc ggctgtgcga acgcattctg gcgggctcga gcggcgcgat
     1681 cgcccacaac accacagtgt tccagggcgt ggcgggccag tccctgcagg tgtcttgccc
     1741 ctatgactcc atgaagcact gggggaggcg caaggcctgg tgccgccagc tgggagagaa
     1801 gggcccatgc cagcgtgtgg tcagcacgca caacttgtgg ctgctgtcct tcctgaggag
     1861 gtggaatggg agcacagcca tcacagacga taccctgggt ggcactctca ccattacgct
     1921 gcggaatcta caaccccatg atgcgggtct ctaccagtgc cagagcctcc atggcagtga
     1981 ggctgacacc ctcaggaagg tcctggtgga ggtgctggca gaccccctgg atcaccggga
     2041 tgctggagat ctctggttcc ccggggagtc tgagagcttc gaggatgccc atgtggagca
     2101 cagcatctcc aggagcctct tggaaggaga aatccccttc ccacccactt ccatccttct
     2161 cctcctggcc tgcatctttc tcatcaagat tctagcagcc agcgccctct gggctgcagc
     2221 ctggcatgga cagaagccag ggacacatcc acccagtgaa ctggactgtg gccatgaccc
     2281 agggtatcag ctccaaactc tgccagggct gagagacacg gtttaaacga attcgggctc
     2341 ggtacccggg gatcctctag agtcgacctg caggcatgca agctgatccg gctgctaaca
     2401 aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc
     2461 ttggggcggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     2521 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     2581 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     2641 caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta caaatgtggt
     2701 aaaatcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt
     2761 acgcatctgt gcggtatttc acaccgcata cgcggatctg cgcagcacca tggcctgaaa
     2821 taacctctga aagaggaact tggttaggta ccttctgagg cggaaagaac cagctgtgga
     2881 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2941 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3001 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3061 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3121 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     3181 gaggcttttt tggaggccta ggcttttgca aaaagcttaa ttaactgttg acaattaatc
     3241 atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa cccaggaggc
     3301 agatcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     3361 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     3421 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     3481 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     3541 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     3601 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     3661 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     3721 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     3781 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     3841 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     3901 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     3961 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4021 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4081 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     4141 caacctgcca tcacgatggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg
     4201 gttttttgtg tgaatcgata gcgataagga tcctctttgc gcttgcgttt tcccttgtcc
     4261 agatagccca gtagctgaca ttcatccggg gtcagcaccg tttctgcgga ctggctttct
     4321 acccggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4381 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4441 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4501 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4561 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4621 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4681 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4741 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4801 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4861 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     4921 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4981 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatttcaag
     5041 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5101 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta tagtccggaa
     5161 atacaggaac gcacgctgga tggcccttcg ctgggatggt gaaaccatga aaaatggcag
     5221 cttcagtgga ttaagtgggg gtaatgtggc ctgtaccctc tggttgcata ggtattcata
     5281 cggttaaaat ttatcaggcg cgattgcggc agtttttcgg gtggtttgtt gccattttta
     5341 cctgtctgct gccgtgatcg cgctgaacgc gttttagcgg tgcgtacaat taagggatta
     5401 tggtaaatcc acttactgtc tgccctcgta gccatcgaga taaaccgcag tactccggcc
     5461 acgatgcgtc cggcgtagag gatcgagatc t
//
