LOCUS       sequence-474906-        4814 bp    DNA     circular SYN 06-JUN-2025
DEFINITION  Express SmBiT-MAFG 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 4814)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 4814)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 6, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: SmBiT-MAFG Fusion Vector
FEATURES             Location/Qualifiers
     source          1..4814
                     /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    1877..1998
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1888..1907)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2113..2146)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2156..2203
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2235..3029
                     /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(2289..2308)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2899..2918
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3187..3775)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3267..3286)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3901..3949
                     /note="synthetic polyadenylation signal"
     CDS             complement(3973..4371)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4368..4377
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4402..4759)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4416..4551
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4568..4759
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4716..4736
                     /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 cgctcaccat ggtgaccggc
     1201 taccggctgt tcgaggagat tctcgggagc tccggtggtg gcgggagcgg aggtggaggc
     1261 tcgagcggtg cgatcgccat gacgaccccc aataaaggaa acaaggcctt gaaggtgaag
     1321 cgggagccgg gtgagaatgg caccagcctg acggatgagg agctggtgac catgtcggtg
     1381 cgggagctga accagcacct gcggggcctg tccaaggagg agatcgtcca gctgaagcag
     1441 cgccggcgca cgctcaagaa ccgcggctac gctgccagct gccgcgtgaa gcgggtgacg
     1501 cagaaggagg agctggagaa gcagaaggcg gagctgcagc aggaggtgga gaagctggcc
     1561 tcagagaacg ccagcatgaa gctggagctc gacgcgctgc gctccaagta cgaggcgctg
     1621 cagaccttcg cccggacggt ggcccgcagc cccgtggcgc cagcccgggg cccccttgcc
     1681 gccggcctgg ggcccctcgt cccaggcaag gtggccgcca ccagcgtcat cacaatagta
     1741 aagtccaaga cggatgcccg atcggtttaa acgaattcgg gctcggtacc cggggatcct
     1801 ctagagtcga cctgcaggca tgcaagctga tttggctgct gccaccgctg agcaaggccg
     1861 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     1921 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     1981 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     2041 gagatgtggg aggttttttt aagcaagtaa aacctctaca aatgtggtaa aatcttaatt
     2101 aagaattcta atataacttc gtatagcata cattatacga agttatggat ccactgttga
     2161 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
     2221 ccaggaggca gatcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     2281 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     2341 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     2401 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     2461 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     2521 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     2581 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     2641 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     2701 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     2761 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     2821 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     2881 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     2941 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     3001 tctatcgcct tcttgacgag ttcttctgat tcgaaatgac cgaccaagcg acgcccaacc
     3061 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3121 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3181 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3241 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3301 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3361 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3421 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3481 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3541 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3601 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     3661 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3721 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga
     3781 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3841 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttatagt ccggaatcgt
     3901 cacacaaaaa accaacacac agatgtaatg aaaataaaga tattttattg cggccatcgt
     3961 gatggctaga gtttagccct cccacacata accagagggc agcaattcac gaatcccaac
     4021 tgccgtcggc tgtccatcac tgtccttcac tatggctttg atcccaggat gcagatcgag
     4081 aagcacctgt cggcaccgtc cgcaggggct caagatgccc ctgttctcat ttccgatcgc
     4141 gacgatacaa gtcaggttgc cagctgccgc agcagcagca gtgcccagca ccacgagttc
     4201 tgcacaaggt cccccagtaa aatgatatac attgacacca gtgaagatgc ggccgtcgct
     4261 agagagagct gcgctggcga cgctgtagtc ttcagagatg gggatgctgt tgattgtagc
     4321 cgttgctctt tcaatgaggg tggattcttc ttgagacaaa ggcttggcca tggtggcgct
     4381 agtgtcagaa gaatcgagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta
     4441 cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag
     4501 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
     4561 taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     4621 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
     4681 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
     4741 ctggttcttt ccgcctcaga aggtacctaa ccaagttcct ctttcagagg ttatttcagg
     4801 ccatggtgct gcgc
//
