LOCUS       sequence-474683-        5276 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express MAFG-LgBiT 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 5276)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5276)
  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            "Unpublished"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-LgBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5276
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    17..98
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     42..61
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(1330..1348)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1331..1348)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        1468..1771
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1926..1946
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        1979..2026
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2058..2852
                     /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(2112..2131)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2722..2741
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3010..3598)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3090..3109)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3724..3772
                     /note="synthetic polyadenylation signal"
     CDS             complement(3796..4833)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3809..3817
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(4864..5221)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4878..5013
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5030..5221
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5178..5198
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtg acgtcggcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
       61 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
      121 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
      181 aatgtggtat ggctgattat gatccgcggc cgcaatacgt cgacgttatc agctgacttc
      241 gtacgagagc ctaggtgttt ctagagtcgc ggccttagct gttgatggtt actcggaaca
      301 gcatggagcc gtcgggggtg atcaggcgct cgtcgataat tttgttgccg ttccacaggg
      361 tccctgttac agtgatcttt ttgccgtcga acacggcgat gccttcatac ggccgtccga
      421 aatagttcag catgttcggc gtaaccccgt cgattaccag tgtgccatag ggcaggatca
      481 ccttaaagtg atgatcatcc acagggtaca ccaccttaaa cacctcttcg atctgggcca
      541 tttggtcggc gctcagacct tcatacggga tgatgacatg gatgtcgatc ttcagggcat
      601 tttcaccgct ccggacaatc ctttggatcg gagttacgga cacggcgaga ttctgcagca
      661 aactggacac acctccctgt tcaaggactt ggtccaggtt gtaggcggct gtctgttccc
      721 agtccccaac gaaatcttcg agtgtgaaga caccgctcga gcctccacct ccgctcccgc
      781 caccacctga actgccttga gaaaccgatc gggcatccgt cttggacttt actattgtga
      841 tgacgctggt ggcggccacc ttgcctggga cgaggggccc caggccggcg gcaagggggc
      901 cccgggctgg cgccacgggg ctgcgggcca ccgtccgggc gaaggtctgc agcgcctcgt
      961 acttggagcg cagcgcgtcg agctccagct tcatgctggc gttctctgag gccagcttct
     1021 ccacctcctg ctgcagctcc gccttctgct tctccagctc ctccttctgc gtcacccgct
     1081 tcacgcggca gctggcagcg tagccgcggt tcttgagcgt gcgccggcgc tgcttcagct
     1141 ggacgatctc ctccttggac aggccccgca ggtgctggtt cagctcccgc accgacatgg
     1201 tcaccagctc ctcatccgtc aggctggtgc cattctcacc cggctcccgc ttcaccttca
     1261 aggccttgtt tcctttattg ggggtcgtca tggcgatcgc tagctgcaaa aagaacaagt
     1321 agcttgtatt ctatagtgtc acctaaatgg gcgatctgac ggttcactaa accagctctg
     1381 cttatatagg cctccctggc gcgccactag ttattaatag taatcaatta cggggtcatt
     1441 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     1501 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     1561 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     1621 ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca atgacggtaa
     1681 atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta cttggcagta
     1741 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acaccaatgg
     1801 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     1861 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata accccgcccc
     1921 gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc aaatcgaatt ctaatactgt
     1981 tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta
     2041 aacccaggag gcagatcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     2101 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     2161 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     2221 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     2281 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     2341 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     2401 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     2461 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     2521 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     2581 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     2641 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     2701 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     2761 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     2821 ccttctatcg ccttcttgac gagttcttct gattcgaaat gaccgaccaa gcgacgccca
     2881 accggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2941 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3001 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3061 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3121 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3181 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3241 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3301 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3361 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3421 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     3481 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3541 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatttcaaga
     3601 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3661 gattttggtc atgagattat caaaaaggat cttcacctag atccttttat agtccggaat
     3721 cgtcacacaa aaaaccaaca cacagatgta atgaaaataa agatatttta ttgcggccat
     3781 cgtgatggct agagtctaaa ctcgacctac ctccttagcg cgcggtcgtg tactgggccg
     3841 gcggttgccg ctgtcggcca gcacctcgac gcagccgtcg gtccatacgg ctgcgctccg
     3901 gcgagcgatt tgtgtgcgac cgacggtgcc ggccccgctg cggacgatgg catcgcagcg
     3961 gccttgagcc caggcagcat cgtcgaagtt gccgtccacg aggctctggt agagctgatc
     4021 caggccgatg cgcagcatgt aggctcgcag acgagggctg ccggccagct cgggatgccg
     4081 gcgctcgaag tagcgagtct gctgctccat gcaagccagc cagggccgcc agaagaagat
     4141 gttggccacc tcgtactgac tgtccccgaa catagcttcg gaccagtcga ttacggcggt
     4201 gatgcggccg ttgtcggtca ggacgttgtt gctgccgaag tcggcatgga cgaggtggcg
     4261 cacctcggga cagtcttcgg cccacagcat cagttcgtcc agggcttgag ctacgctggc
     4321 ggacacggtg tcgtccatca cggtctgcca gtggtagaca tggggatcag caatggcgca
     4381 aatgaaatcc cgccaggttg tgtactggcc gatgccttgg ggcccgaatg ggccgaagcc
     4441 gctggtttga ctgaggtcgg cggcggcaat agcatccata gcttcggcga caggctgtaa
     4501 cacagcaggc agctctgttt cggggaggtc ttggagagta acgccttggg cgcgtctact
     4561 gatgcagtat gtcaggctct cgctgaactc gccgatgtcc aacacttcgg ggatgggtag
     4621 tgcagcgctg gcgaagtggc ggtacacgta gcggtctttg tagaagccat cagcgcagct
     4681 attcacccgc agtacatagc cgcgtccgcc gacatcgaag ctgaaggctc ggctctcttc
     4741 gccctccgac aactgcatca ggtcgctcac actgtcgaac ttctcgatga gaaatttttc
     4801 aacgctggta gcggtgagtt cgggcttctt catggtggcg ctagtgtcag aagaatcgag
     4861 ctttttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga
     4921 ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga
     4981 atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt
     5041 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     5101 tccacacctg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     5161 gcctggggac tttccacacc ctaactgaca cacattccac agctggttct ttccgcctca
     5221 gaaggtacct aaccaagttc ctctttcaga ggttatttca ggccatggtg ctgcgc
//
