LOCUS       sequence-479914-        4815 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express DAP12-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 4815)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4815)
  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            "Unpublished"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: DAP12-SmBiT BiBiT Ready Vector
FEATURES             Location/Qualifiers
     source          1..4815
                     /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    1704..1825
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1715..1734)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(1940..1973)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1983..2030
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2062..2856
                     /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(2116..2135)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2726..2745
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      3073..3618
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     3519..3538
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3735..3752
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3902..3950
                     /note="synthetic polyadenylation signal"
     CDS             complement(3974..4372)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4369..4378
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4403..4760)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4417..4552
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4569..4760
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4717..4737
                     /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 ggggggactt
     1201 gaaccctgca gcaggctcct gctcctgcct ctcctgctgg ctgtaagtgg tctccgtcct
     1261 gtccaggccc aggcccagag cgattgcagt tgctctacgg tgagcccggg cgtgctggca
     1321 gggatcgtga tgggagacct ggtgctgaca gtgctcattg ccctggccgt gtacttcctg
     1381 ggccggctgg tccctcgggg gcgaggggct gcggaggcag cgacccggaa acagcgtatc
     1441 actgagaccg agtcgcctta tcaggagctc cagggtcaga ggtcggatgt ctacagcgac
     1501 ctcaacacac agaggccgta ttacaaagtt tctcaaggca gttcaggtgg tggcgggagc
     1561 ggaggtggag gctcgagcgg tgtgaccggc taccggctgt tcgaggagat tctgtaaggc
     1621 cgcgactcta gagtcgacct gcaggcatgc aagctgattt ggctgctgcc accgctgagc
     1681 aaggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     1741 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     1801 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     1861 tcagggggag atgtgggagg tttttttaag caagtaaaac ctctacaaat gtggtaaaat
     1921 cttaattaag aattctaata taacttcgta tagcatacat tatacgaagt tatggatcca
     1981 ctgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
     2041 actaaaccca ggaggcagat catgattgaa caagatggat tgcacgcagg ttctccggcc
     2101 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     2161 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     2221 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     2281 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     2341 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     2401 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     2461 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     2521 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     2581 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     2641 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     2701 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     2761 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     2821 atcgccttct atcgccttct tgacgagttc ttctgattcg aaatgaccga ccaagcgacg
     2881 cccaaccggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2941 acgcaggaaa gaacatgtca aacatgagaa ttacaactta tatcgtatgg ggctgacttc
     3001 aggtgctaca tttgaagaga taaattgcac tgaaatctag aaatatttta tctgattaat
     3061 aagatgatct tcttgagatc gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa
     3121 aaccgccttg cagggcggtt tttcgaaggt tctctgagct accaactctt tgaaccgagg
     3181 taactggctt ggaggagcgc agtcaccaaa acttgtcctt tcagtttagc cttaaccggc
     3241 gcatgacttc aagactaact cctctaaatc aattaccagt ggctgctgcc agtggtgctt
     3301 ttgcatgtct ttccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3361 actgaacggg gggttcgtgc atacagtcca gcttggagcg aactgcctac ccggaactga
     3421 gtgtcaggcg tggaatgaga caaacgcggc cataacagcg gaatgacacc ggtaaaccga
     3481 aaggcaggaa caggagagcg cacgagggag ccgccagggg gaaacgcctg gtatctttat
     3541 agtcctgtcg ggtttcgcca ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg
     3601 gggcggagcc tatggaaaaa cggctttgcc gcggccctct cacttccctg ttaagtatct
     3661 tcctggcatc ttccaggaaa tctccgcccc gttcgtaagc catttccgct cgccgcagtc
     3721 gaacgaccga gcgtagcgag tcagtgagcg aggaagcgga atatatcctg tatcacatat
     3781 tctgctgacg caccggtgca gccttttttc tcctgccaca tgaagcactt cactgacacc
     3841 ctcatcagtg ccaacatagt aagccagtat acactccgct agcgctgatg tccggaatcg
     3901 tcacacaaaa aaccaacaca cagatgtaat gaaaataaag atattttatt gcggccatcg
     3961 tgatggctag agtttagccc tcccacacat aaccagaggg cagcaattca cgaatcccaa
     4021 ctgccgtcgg ctgtccatca ctgtccttca ctatggcttt gatcccagga tgcagatcga
     4081 gaagcacctg tcggcaccgt ccgcaggggc tcaagatgcc cctgttctca tttccgatcg
     4141 cgacgataca agtcaggttg ccagctgccg cagcagcagc agtgcccagc accacgagtt
     4201 ctgcacaagg tcccccagta aaatgatata cattgacacc agtgaagatg cggccgtcgc
     4261 tagagagagc tgcgctggcg acgctgtagt cttcagagat ggggatgctg ttgattgtag
     4321 ccgttgctct ttcaatgagg gtggattctt cttgagacaa aggcttggcc atggtggcgc
     4381 tagtgtcaga agaatcgagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact
     4441 acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta
     4501 gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag
     4561 ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
     4621 ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca
     4681 tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac acattccaca
     4741 gctggttctt tccgcctcag aaggtaccta accaagttcc tctttcagag gttatttcag
     4801 gccatggtgc tgcgc
//
