LOCUS       sequence-471887-        4923 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express CD3E-SmBiT 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 4923)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4923)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: CD3E-SmBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..4923
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    24..105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        338..641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        642..845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     796..816
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          981..1113
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1096..1115
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1157..1176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1157..1175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1986..2107
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1997..2016)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2222..2255)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2265..2312
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2344..3138
                     /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(2398..2417)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3008..3027
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3296..3884)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3376..3395)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4010..4058
                     /note="synthetic polyadenylation signal"
     CDS             complement(4082..4480)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4477..4486
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4511..4868)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4525..4660
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4677..4868
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4825..4845
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtt aattaagacg tcggcagtga aaaaaatgct ttatttgtga aatttgtgat
       61 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
      121 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
      181 ctctacaaat gtggtatggc tgattatgat ccgcggccgc aatacgtcga cgttatcagc
      241 tgacttcgta cgagagccta ggattatggc gcgccactag ttattaatag taatcaatta
      301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
      541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
      601 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      661 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      721 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
      781 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      841 gagctggttt agtgaaccgt cagatcacta gaagctttat tgcggtagtt tatcacagtt
      901 aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt
      961 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1021 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1081 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     1141 cttaaggcta gagtattaat acgactcact atagggctag cgatcgccat gcagtcgggc
     1201 actcactgga gagttctggg cctctgcctc ttatcagttg gcgtttgggg gcaagatggt
     1261 aatgaagaaa tgggtggtat tacacagaca ccatataaag tctccatctc tggaaccaca
     1321 gtaatattga catgccctca gtatcctgga tctgaaatac tatggcaaca caatgataaa
     1381 aacataggcg gtgatgagga tgataaaaac ataggcagtg atgaggatca cctgtcactg
     1441 aaggaatttt cagaattgga gcaaagtggt tattatgtct gctaccccag aggaagcaaa
     1501 ccagaagatg cgaactttta tctctacctg agggcaagag tgtgtgagaa ctgcatggag
     1561 atggatgtga tgtcggtggc cacaattgtc atagtggaca tctgcatcac tgggggcttg
     1621 ctgctgctgg tttactactg gagcaagaat agaaaggcca aggccaagcc tgtgacacga
     1681 ggagcgggtg ctggcggcag gcaaagggga caaaacaagg agaggccacc acctgttccc
     1741 aacccagact atgagcccat ccggaaaggc cagcgggacc tgtattctgg cctgaatcag
     1801 agacgcatcg tttctcaagg cagttcaggt ggtggcggga gcggaggtgg aggctcgagc
     1861 ggtgtgaccg gctaccggct gttcgaggag attctgtaag gccgcgactc tagagtcgac
     1921 ctgcaggcat gcaagctgat ttggctgctg ccaccgctga gcaaggccgc ttcgagcaga
     1981 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     2041 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     2101 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga
     2161 ggttttttta agcaagtaaa acctctacaa atgtggtaaa atcttaatta agaattctaa
     2221 tataacttcg tatagcatac attatacgaa gttatggatc cactgttgac aattaatcat
     2281 cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc caggaggcag
     2341 atcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     2401 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     2461 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     2521 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     2581 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     2641 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     2701 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     2761 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     2821 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     2881 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     2941 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     3001 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     3061 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     3121 cttgacgagt tcttctgatt cgaaatgacc gaccaagcga cgcccaaccg gtatcagctc
     3181 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3241 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3301 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3361 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3421 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3481 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3541 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3601 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3661 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3721 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3781 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3841 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt tcaagaagat cctttgatct
     3901 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3961 gattatcaaa aaggatcttc acctagatcc ttttatagtc cggaatcgtc acacaaaaaa
     4021 ccaacacaca gatgtaatga aaataaagat attttattgc ggccatcgtg atggctagag
     4081 tttagccctc ccacacataa ccagagggca gcaattcacg aatcccaact gccgtcggct
     4141 gtccatcact gtccttcact atggctttga tcccaggatg cagatcgaga agcacctgtc
     4201 ggcaccgtcc gcaggggctc aagatgcccc tgttctcatt tccgatcgcg acgatacaag
     4261 tcaggttgcc agctgccgca gcagcagcag tgcccagcac cacgagttct gcacaaggtc
     4321 ccccagtaaa atgatataca ttgacaccag tgaagatgcg gccgtcgcta gagagagctg
     4381 cgctggcgac gctgtagtct tcagagatgg ggatgctgtt gattgtagcc gttgctcttt
     4441 caatgagggt ggattcttct tgagacaaag gcttggccat ggtggcgcta gtgtcagaag
     4501 aatcgagctt tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata
     4561 gctcagaggc cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg
     4621 gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga
     4681 ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
     4741 gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
     4801 ctggggagcc tggggacttt ccacacccta actgacacac attccacagc tggttctttc
     4861 cgcctcagaa ggtacctaac caagttcctc tttcagaggt tatttcaggc catggtgctg
     4921 cgc
//
