LOCUS       sequence-471894-        5816 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express SmBiT-MDM2 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 5816)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5816)
  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            "Unpublished"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: SmBiT-MDM2 Control Vector
FEATURES             Location/Qualifiers
     source          1..5816
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        80..271
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     228..248
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    516..637
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(553..572)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     607..626
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     674..692
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     primer_bind     complement(2399..2418)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2400..2418)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2460..2479)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          2462..2594
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(2730..2933)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2759..2779)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2934..3313
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    3504..3787
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     rep_origin      3903..4491
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4392..4411
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4730..4778
                     /note="synthetic polyadenylation signal"
     CDS             complement(4842..5636)
                     /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(4953..4972)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5563..5582
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5668..5715)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(5730..5816,1..271))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5744..5816,1..63)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg
       61 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat
      121 acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg
      181 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca
      241 cacattccac agctggttct ttccgcctca gaaggtacct aaccaagttc ctctttcaga
      301 ggttatttca ggccatggtg ctgcgcagat ccgcgtatgc ggtgtgaaat accgcacaga
      361 tgcgtaagga gaaaataccg catcaggaaa ttgtaagcgt taatattttg ttaaaattcg
      421 attttaccac atttgtagag gttttacttg cttaaaaaaa cctcccacat ctccccctga
      481 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
      541 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
      601 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgctcg aagcggccgc
      661 cccaaggggt tatgctagtt attgctcagc ggtggcagca gccaactcag cttcctttcg
      721 ggctttgtta gcagccggat cagcttgcat gcctgcaggt cgactctaga ggatccccgg
      781 gtaccgagcc cgaattcgtt taaacgggga aataagttag cacaatcatt tgaattggtt
      841 gtctacatac tgggcagggc ttattccttt tctttagctt ctttgcacat gtaaagcagg
      901 ccataagatg tcctgttttg ccatggacaa tgcaaccatt tttaggtcga ccttgacaaa
      961 tcacacaagg ttcaatggca ttaaggggca aactagattc cacactctct tctttgtctt
     1021 gggtttcttc cctttcaaac tctttcacat cttcttggct gctataaata atgctactag
     1081 aagttgatgg ctgagaatag tcttcacttt cttgtgattg tgaagcttgt gtaattttat
     1141 catcattttc ctcaacacat gactctctgg aatcattcac tatagttttt ttacaatcag
     1201 gaacatcaaa gccctcttca gcttgtgttg agttttccag tttggctttc tcagagattt
     1261 cccctttatc tttcccttta tcttcaggaa gccaattctc acgaagggcc caacatctgt
     1321 tgcaatgtga tggaaggggg ggattcattt cattgcatga agtgcatttc caatagtcag
     1381 ctaaggaaat ttcaggatct tcttcaaatg aatctgtatc actctcccct gcctgataca
     1441 cagtaacttg atatacctca tcatcttcat ctgagagttc ttgtccttct tcactaaggc
     1501 tataatcttc tgagtcgaga gattcaactt caaattctac actaaactga tctgaaactg
     1561 aatcctgatc caaccaatca cctgaatgtt cacttacacc agcatcaaga tccggattcg
     1621 atggcgtccc tgtagattca ctgctactgc ttctttcaca acatatctcc cttattacac
     1681 acagagccag gctttcatca aaggaaaggg aaatactatc agatttgtgg cgttttcttt
     1741 gtcgttcacc agataattca tctgaatttt cttctgtctc actaattgct ctccttctag
     1801 atgaggtaga tggtctagaa accaaatgtg aagatgaagg tttctcttcc tgaagctctt
     1861 gtacaaggtc cttttgatca ctcccacctt caaggtgaca cctgttctca ctcacagatg
     1921 tacctgagtc cgatgattcc tgctgattga ctactaccaa gttcctgtag atcatggtat
     1981 atattttcct gtgctctttc acagagaagc ttggcacgcc aaacaaatct cctagaagat
     2041 catttgaaca atatacaata tgttgttgct tctcatcata taatcgttta gtcataatat
     2101 actggccaag ataaaaaaga acctctttca tagtataagt gtctttttgt gcaccaacag
     2161 actttaataa cttcaaaagc aatggctttg gtctaaccag ggtctcttgt tccgaagctg
     2221 gaatctgtga ggtggttaca gcaccatcag taggtacaga catgttggta ttgcacattt
     2281 gcctgctcct caccatggcg atcgcaccgc tcgagcctcc acctccgctc ccgccaccac
     2341 cggagctccc gagaatctcc tcgaacagcc ggtagccggt caccatggtg agcgctagcc
     2401 ctatagtgag tcgtattaat actctagcct taagagctgt aattgaactg ggagtggaca
     2461 cctgtggaga gaaaggcaaa gtggatgtca gtaagaccaa taggtgccta tcagaaacgc
     2521 aagagtcttc tctgtctcga caagcccagt ttctattggt ctccttaaac ctgtcttgta
     2581 accttgatac ttacctgccc agtgcctcac gaccaacttc tgcagcttaa gttcgagact
     2641 gttgtgtcag aagcactgac tgcgttagca atttaactgt gataaactac cgcaataaag
     2701 cttctagtga tctgacggtt cactaaacca gctctgctta tatagacctc ccaccgtaca
     2761 cgcctaccgc ccatttgcgt caacggggcg gggttattac gacattttgg aaagtcccgt
     2821 tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc
     2881 cgtgagtcaa accgctatcc acgcccattg gtgtactgcc aaaaccgcat caccatggta
     2941 atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccgtaa ggtcatgtac
     3001 tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg cggacttggc
     3061 atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga
     3121 cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat
     3181 gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg
     3241 aactccatat atgggctatg aactaatgac cccgtaattg attactatta ataactagtc
     3301 aataatcaat gccaacatgg cggtcatatt ggacatgagc caatataaat gtacatatta
     3361 tgatatagat acaacgtatg caatggccaa tagccaatat tgatttatgc tatataacca
     3421 atgaataata tggctaatgg ccaatattga agatctcgat cctctacgcc ggacgcatcg
     3481 tggccggagt actgcggttt atctcgatgg ctacgagggc agacagtaag tggatttacc
     3541 ataatccctt aattgtacgc accgctaaaa cgcgttcagc gcgatcacgg cagcagacag
     3601 gtaaaaatgg caacaaacca cccgaaaaac tgccgcaatc gcgcctgata aattttaacc
     3661 gtatgaatac ctatgcaacc agagggtaca ggccacatta cccccactta atccactgaa
     3721 gctgccattt ttcatggttt caccatccca gcgaagggcc atccagcgtg cgttcctgta
     3781 tttccggact ataaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     3841 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3901 tcttgaaatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3961 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4021 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     4081 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4141 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4201 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4261 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4321 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4381 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4441 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4501 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     4561 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccggg
     4621 tagaaagcca gtccgcagaa acggtgctga ccccggatga atgtcagcta ctgggctatc
     4681 tggacaaggg aaaacgcaag cgcaaagagg atccttatcg ctatcgattc acacaaaaaa
     4741 ccaacacaca gatgtaatga aaataaagat attttattgc ggccatcgtg atggcaggtt
     4801 gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga
     4861 aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag
     4921 cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc
     4981 tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt
     5041 tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg
     5101 ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg
     5161 tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga
     5221 tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt
     5281 gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc
     5341 cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca
     5401 gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcctgcagt
     5461 tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac
     5521 agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat
     5581 agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgatc
     5641 tgcctcctgg gtttagttcc tcaccttgtc gtattatact atgccgatat actatgccga
     5701 tgattaattg tcaacagtta attaagcttt ttgcaaaagc ctaggcctcc aaaaaagcct
     5761 cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaat
//
