LOCUS       sequence-479791-        5853 bp    DNA     circular SYN 05-AUG-2025
DEFINITION  Express TREM1-HaloTag 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 5853)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5853)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 5, 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            "Unpublished"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-HaloTag
FEATURES             Location/Qualifiers
     source          1..5853
                     /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..1808
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1824..2708
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     primer_bind     complement(2796..2814)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2851..2972
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2862..2881)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2916..2935
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3217..3574
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3217..3408
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3240..3260)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3425..3560
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3589..3636
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3668..4462
                     /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(3722..3741)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4332..4351
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4526..4574
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4813..5401)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4893..4912)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5517..5800
                     /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 ctctcgagcc aaccactgag gatctgtact
     1801 ttcagagcga taacgatgga tccgaaatcg gtactggctt tccattcgac ccccattatg
     1861 tggaagtcct gggcgagcgc atgcactacg tcgatgttgg tccgcgcgat ggcacccctg
     1921 tgctgttcct gcacggtaac ccgacctcct cctacgtgtg gcgcaacatc atcccgcatg
     1981 ttgcaccgac ccatcgctgc attgctccag acctgatcgg tatgggcaaa tccgacaaac
     2041 cagacctggg ttatttcttc gacgaccacg tccgcttcat ggatgccttc atcgaagccc
     2101 tgggtctgga agaggtcgtc ctggtcattc acgactgggg ctccgctctg ggtttccact
     2161 gggccaagcg caatccagag cgcgtcaaag gtattgcatt tatggagttc atccgcccta
     2221 tcccgacctg ggacgaatgg ccagaatttg cccgcgagac cttccaggcc ttccgcacca
     2281 ccgacgtcgg ccgcaagctg atcatcgatc agaacgtttt tatcgagggt acgctgccga
     2341 tgggtgtcgt ccgcccgctg actgaagtcg agatggacca ttaccgcgag ccgttcctga
     2401 atcctgttga ccgcgagcca ctgtggcgct tcccaaacga gctgccaatc gccggtgagc
     2461 cagcgaacat cgtcgcgctg gtcgaagaat acatggactg gctgcaccag tcccctgtcc
     2521 cgaagctgct gttctggggc accccaggcg ttctgatccc accggccgaa gccgctcgcc
     2581 tggccaaaag cctgcctaac tgcaaggctg tggacatcgg cccgggtctg aatctgctgc
     2641 aagaagacaa cccggacctg atcggcagcg agatcgcgcg ctggctgtct actctggaga
     2701 tttccggtta atagaattct agagtcgacc tgcaggcatg caagctgatc cggctgctaa
     2761 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
     2821 ccttggggcg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     2881 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     2941 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     3001 ttcaggttca gggggagatg tgggaggttt ttttaagcaa gtaaaacctc tacaaatgtg
     3061 gtaaaatcga attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc
     3121 ttacgcatct gtgcggtatt tcacaccgca tacgcggatc tgcgcagcac catggcctga
     3181 aataacctct gaaagaggaa cttggttagg taccttctga ggcggaaaga accagctgtg
     3241 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3301 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3361 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3421 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3481 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     3541 aggaggcttt tttggaggcc taggcttttg caaaaagctt aattaactgt tgacaattaa
     3601 tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta aacccaggag
     3661 gcagatcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3721 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3781 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3841 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3901 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3961 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4021 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4081 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4141 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     4201 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4261 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4321 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4381 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4441 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4501 cccaacctgc catcacgatg gccgcaataa aatatcttta ttttcattac atctgtgtgt
     4561 tggttttttg tgtgaatcga tagcgataag gatcctcttt gcgcttgcgt tttcccttgt
     4621 ccagatagcc cagtagctga cattcatccg gggtcagcac cgtttctgcg gactggcttt
     4681 ctacccggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4741 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4801 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4861 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4921 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4981 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5041 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5101 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5161 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5221 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     5281 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5341 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatttca
     5401 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5461 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt tatagtccgg
     5521 aaatacagga acgcacgctg gatggccctt cgctgggatg gtgaaaccat gaaaaatggc
     5581 agcttcagtg gattaagtgg gggtaatgtg gcctgtaccc tctggttgca taggtattca
     5641 tacggttaaa atttatcagg cgcgattgcg gcagtttttc gggtggtttg ttgccatttt
     5701 tacctgtctg ctgccgtgat cgcgctgaac gcgttttagc ggtgcgtaca attaagggat
     5761 tatggtaaat ccacttactg tctgccctcg tagccatcga gataaaccgc agtactccgg
     5821 ccacgatgcg tccggcgtag aggatcgaga tct
//
