LOCUS       sequence-479820-        5486 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express LgBiT-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 5486)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5486)
  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            "Unpublished"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: LgBiT-TREM2 Vector
FEATURES             Location/Qualifiers
     source          1..5486
                     /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"
     primer_bind     complement(2429..2447)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2484..2605
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2495..2514)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2549..2568
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2850..3207
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2850..3041
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2873..2893)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3058..3193
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3222..3269
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3301..4095
                     /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(3355..3374)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3965..3984
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4159..4207
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4446..5034)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4526..4545)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5150..5433
                     /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 ggctagcgct caccatgaac tccttctcca
     1081 caagcgcctt cggtccagtt gccttctccc tgggcctgct cctggtgttg cctgctgcct
     1141 tccctgcccc agtcttcaca ctcgaagatt tcgttgggga ctgggaacag acagccgcct
     1201 acaacctgga ccaagtcctt gaacagggag gtgtgtccag tttgctgcag aatctcgccg
     1261 tgtccgtaac tccgatccaa aggattgtcc ggagcggtga aaatgccctg aagatcgaca
     1321 tccatgtcat catcccgtat gaaggtctga gcgccgacca aatggcccag atcgaagagg
     1381 tgtttaaggt ggtgtaccct gtggatgatc atcactttaa ggtgatcctg ccctatggca
     1441 cactggtaat cgacggggtt acgccgaaca tgctgaacta tttcggacgg ccgtatgaag
     1501 gcatcgccgt gttcgacggc aaaaagatca ctgtaacagg gaccctgtgg aacggcaaca
     1561 aaattatcga cgagcgcctg atcacccccg acggctccat gctgttccga gtaaccatca
     1621 acagtgggag ctccggtggt ggcgggagcg gaggtggagg ctcgagcggt gcgatcgccc
     1681 acaacaccac agtgttccag ggcgtggcgg gccagtccct gcaggtgtct tgcccctatg
     1741 actccatgaa gcactggggg aggcgcaagg cctggtgccg ccagctggga gagaagggcc
     1801 catgccagcg tgtggtcagc acgcacaact tgtggctgct gtccttcctg aggaggtgga
     1861 atgggagcac agccatcaca gacgataccc tgggtggcac tctcaccatt acgctgcgga
     1921 atctacaacc ccatgatgcg ggtctctacc agtgccagag cctccatggc agtgaggctg
     1981 acaccctcag gaaggtcctg gtggaggtgc tggcagaccc cctggatcac cgggatgctg
     2041 gagatctctg gttccccggg gagtctgaga gcttcgagga tgcccatgtg gagcacagca
     2101 tctccaggag cctcttggaa ggagaaatcc ccttcccacc cacttccatc cttctcctcc
     2161 tggcctgcat ctttctcatc aagattctag cagccagcgc cctctgggct gcagcctggc
     2221 atggacagaa gccagggaca catccaccca gtgaactgga ctgtggccat gacccagggt
     2281 atcagctcca aactctgcca gggctgagag acacggttta aacgaattcg ggctcggtac
     2341 ccggggatcc tctagagtcg acctgcaggc atgcaagctg atccggctgc taacaaagcc
     2401 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
     2461 gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     2521 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     2581 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     2641 tcagggggag atgtgggagg tttttttaag caagtaaaac ctctacaaat gtggtaaaat
     2701 cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca
     2761 tctgtgcggt atttcacacc gcatacgcgg atctgcgcag caccatggcc tgaaataacc
     2821 tctgaaagag gaacttggtt aggtaccttc tgaggcggaa agaaccagct gtggaatgtg
     2881 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2941 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3001 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3061 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3121 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3181 ttttttggag gcctaggctt ttgcaaaaag cttaattaac tgttgacaat taatcatcgg
     3241 catagtatat cggcatagta taatacgaca aggtgaggaa ctaaacccag gaggcagatc
     3301 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     3361 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     3421 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     3481 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     3541 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     3601 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     3661 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     3721 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     3781 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     3841 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     3901 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     3961 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4021 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4081 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     4141 tgccatcacg atggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt
     4201 ttgtgtgaat cgatagcgat aaggatcctc tttgcgcttg cgttttccct tgtccagata
     4261 gcccagtagc tgacattcat ccggggtcag caccgtttct gcggactggc tttctacccg
     4321 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4381 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4441 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4501 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4561 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4621 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4681 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4741 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4801 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4861 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     4921 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4981 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt tcaagaagat
     5041 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5101 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttatagtc cggaaataca
     5161 ggaacgcacg ctggatggcc cttcgctggg atggtgaaac catgaaaaat ggcagcttca
     5221 gtggattaag tgggggtaat gtggcctgta ccctctggtt gcataggtat tcatacggtt
     5281 aaaatttatc aggcgcgatt gcggcagttt ttcgggtggt ttgttgccat ttttacctgt
     5341 ctgctgccgt gatcgcgctg aacgcgtttt agcggtgcgt acaattaagg gattatggta
     5401 aatccactta ctgtctgccc tcgtagccat cgagataaac cgcagtactc cggccacgat
     5461 gcgtccggcg tagaggatcg agatct
//
