LOCUS       sequence-474905-        4788 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express MAFG-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 4788)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4788)
  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            "Unpublished"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: MAFG-SmBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..4788
                     /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    1851..1972
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1862..1881)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2087..2120)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2130..2177
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2209..3003
                     /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(2263..2282)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2873..2892
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3161..3749)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3241..3260)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3875..3923
                     /note="synthetic polyadenylation signal"
     CDS             complement(3947..4345)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4342..4351
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4376..4733)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4390..4525
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4542..4733
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4690..4710
                     /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 gacgaccccc
     1201 aataaaggaa acaaggcctt gaaggtgaag cgggagccgg gtgagaatgg caccagcctg
     1261 acggatgagg agctggtgac catgtcggtg cgggagctga accagcacct gcggggcctg
     1321 tccaaggagg agatcgtcca gctgaagcag cgccggcgca cgctcaagaa ccgcggctac
     1381 gctgccagct gccgcgtgaa gcgggtgacg cagaaggagg agctggagaa gcagaaggcg
     1441 gagctgcagc aggaggtgga gaagctggcc tcagagaacg ccagcatgaa gctggagctc
     1501 gacgcgctgc gctccaagta cgaggcgctg cagaccttcg cccggacggt ggcccgcagc
     1561 cccgtggcgc cagcccgggg cccccttgcc gccggcctgg ggcccctcgt cccaggcaag
     1621 gtggccgcca ccagcgtcat cacaatagta aagtccaaga cggatgcccg atcggtttct
     1681 caaggcagtt caggtggtgg cgggagcgga ggtggaggct cgagcggtgt gaccggctac
     1741 cggctgttcg aggagattct gtaaggccgc gactctagag tcgacctgca ggcatgcaag
     1801 ctgatttggc tgctgccacc gctgagcaag gccgcttcga gcagacatga taagatacat
     1861 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     1921 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     1981 caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt ttttaagcaa
     2041 gtaaaacctc tacaaatgtg gtaaaatctt aattaagaat tctaatataa cttcgtatag
     2101 catacattat acgaagttat ggatccactg ttgacaatta atcatcggca tagtatatcg
     2161 gcatagtata atacgacaag gtgaggaact aaacccagga ggcagatcat gattgaacaa
     2221 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     2281 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     2341 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     2401 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     2461 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     2521 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     2581 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     2641 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     2701 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     2761 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     2821 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     2881 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     2941 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     3001 tgattcgaaa tgaccgacca agcgacgccc aaccggtatc agctcactca aaggcggtaa
     3061 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3121 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3181 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3241 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3301 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3361 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3421 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3481 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3541 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3601 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3661 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3721 agattacgcg cagaaaaaaa ggatttcaag aagatccttt gatcttttct acggggtctg
     3781 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3841 tcttcaccta gatcctttta tagtccggaa tcgtcacaca aaaaaccaac acacagatgt
     3901 aatgaaaata aagatatttt attgcggcca tcgtgatggc tagagtttag ccctcccaca
     3961 cataaccaga gggcagcaat tcacgaatcc caactgccgt cggctgtcca tcactgtcct
     4021 tcactatggc tttgatccca ggatgcagat cgagaagcac ctgtcggcac cgtccgcagg
     4081 ggctcaagat gcccctgttc tcatttccga tcgcgacgat acaagtcagg ttgccagctg
     4141 ccgcagcagc agcagtgccc agcaccacga gttctgcaca aggtccccca gtaaaatgat
     4201 atacattgac accagtgaag atgcggccgt cgctagagag agctgcgctg gcgacgctgt
     4261 agtcttcaga gatggggatg ctgttgattg tagccgttgc tctttcaatg agggtggatt
     4321 cttcttgaga caaaggcttg gccatggtgg cgctagtgtc agaagaatcg agctttttgc
     4381 aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg
     4441 cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg
     4501 aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact
     4561 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
     4621 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
     4681 actttccaca ccctaactga cacacattcc acagctggtt ctttccgcct cagaaggtac
     4741 ctaaccaagt tcctctttca gaggttattt caggccatgg tgctgcgc
//
