LOCUS       sequence-470646-        5273 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 5273)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5273)
  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            "Unpublished"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..5273
                     /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(1327..1345)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1328..1345)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        1465..1768
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1923..1943
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        1976..2023
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2055..2849
                     /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(2109..2128)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2719..2738
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3007..3595)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3087..3106)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3721..3769
                     /note="synthetic polyadenylation signal"
     CDS             complement(3793..4830)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3806..3814
                     /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(4861..5218)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4875..5010
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5027..5218
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5175..5195
                     /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 gccgatcggg catccgtctt ggactttact attgtgatga
      841 cgctggtggc ggccaccttg cctgggacga ggggccccag gccggcggca agggggcccc
      901 gggctggcgc cacggggctg cgggccaccg tccgggcgaa ggtctgcagc gcctcgtact
      961 tggagcgcag cgcgtcgagc tccagcttca tgctggcgtt ctctgaggcc agcttctcca
     1021 cctcctgctg cagctccgcc ttctgcttct ccagctcctc cttctgcgtc acccgcttca
     1081 cgcggcagct ggcagcgtag ccgcggttct tgagcgtgcg ccggcgctgc ttcagctgga
     1141 cgatctcctc cttggacagg ccccgcaggt gctggttcag ctcccgcacc gacatggtca
     1201 ccagctcctc atccgtcagg ctggtgccat tctcacccgg ctcccgcttc accttcaagg
     1261 ccttgtttcc tttattgggg gtcgtcatgg cgatcgctag ctgcaaaaag aacaagtagc
     1321 ttgtattcta tagtgtcacc taaatgggcg atctgacggt tcactaaacc agctctgctt
     1381 atataggcct ccctggcgcg ccactagtta ttaatagtaa tcaattacgg ggtcattagt
     1441 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     1501 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     1561 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     1621 agtacatcaa gtgtatcata tgccaagtcc gccccctatt gacgtcaatg acggtaaatg
     1681 gcccgcctgg cattatgccc agtacatgac cttacgggac tttcctactt ggcagtacat
     1741 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca ccaatgggcg
     1801 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     1861 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaataacc ccgccccgtt
     1921 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtcaaa tcgaattcta atactgttga
     1981 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
     2041 ccaggaggca gatcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     2101 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     2161 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     2221 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     2281 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     2341 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     2401 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     2461 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     2521 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     2581 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     2641 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     2701 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     2761 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     2821 tctatcgcct tcttgacgag ttcttctgat tcgaaatgac cgaccaagcg acgcccaacc
     2881 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2941 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3001 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3061 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3121 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3181 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3241 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3301 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3361 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3421 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     3481 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3541 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga
     3601 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3661 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttatagt ccggaatcgt
     3721 cacacaaaaa accaacacac agatgtaatg aaaataaaga tattttattg cggccatcgt
     3781 gatggctaga gtctaaactc gacctacctc cttagcgcgc ggtcgtgtac tgggccggcg
     3841 gttgccgctg tcggccagca cctcgacgca gccgtcggtc catacggctg cgctccggcg
     3901 agcgatttgt gtgcgaccga cggtgccggc cccgctgcgg acgatggcat cgcagcggcc
     3961 ttgagcccag gcagcatcgt cgaagttgcc gtccacgagg ctctggtaga gctgatccag
     4021 gccgatgcgc agcatgtagg ctcgcagacg agggctgccg gccagctcgg gatgccggcg
     4081 ctcgaagtag cgagtctgct gctccatgca agccagccag ggccgccaga agaagatgtt
     4141 ggccacctcg tactgactgt ccccgaacat agcttcggac cagtcgatta cggcggtgat
     4201 gcggccgttg tcggtcagga cgttgttgct gccgaagtcg gcatggacga ggtggcgcac
     4261 ctcgggacag tcttcggccc acagcatcag ttcgtccagg gcttgagcta cgctggcgga
     4321 cacggtgtcg tccatcacgg tctgccagtg gtagacatgg ggatcagcaa tggcgcaaat
     4381 gaaatcccgc caggttgtgt actggccgat gccttggggc ccgaatgggc cgaagccgct
     4441 ggtttgactg aggtcggcgg cggcaatagc atccatagct tcggcgacag gctgtaacac
     4501 agcaggcagc tctgtttcgg ggaggtcttg gagagtaacg ccttgggcgc gtctactgat
     4561 gcagtatgtc aggctctcgc tgaactcgcc gatgtccaac acttcgggga tgggtagtgc
     4621 agcgctggcg aagtggcggt acacgtagcg gtctttgtag aagccatcag cgcagctatt
     4681 cacccgcagt acatagccgc gtccgccgac atcgaagctg aaggctcggc tctcttcgcc
     4741 ctccgacaac tgcatcaggt cgctcacact gtcgaacttc tcgatgagaa atttttcaac
     4801 gctggtagcg gtgagttcgg gcttcttcat ggtggcgcta gtgtcagaag aatcgagctt
     4861 tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc
     4921 cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg
     4981 ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc
     5041 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     5101 acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     5161 tggggacttt ccacacccta actgacacac attccacagc tggttctttc cgcctcagaa
     5221 ggtacctaac caagttcctc tttcagaggt tatttcaggc catggtgctg cgc
//
