LOCUS       sequence-471866-        6260 bp    DNA     circular SYN 09-MAY-2025
DEFINITION  Express NanoLuc(R)-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 6260)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6260)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 9, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: NanoLuc-MDM2 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1065..1577
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(3203..3221)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3258..3379
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3269..3288)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3323..3342
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3624..3981
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3624..3815
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3647..3667)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3832..3967
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3996..4043
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4075..4869
                     /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(4129..4148)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4739..4758
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4933..4981
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5220..5808)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5300..5319)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5924..6207
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcgct caccatggtc ttcacactcg
     1081 aagatttcgt tggggactgg cgacagacag ccggctacaa cctggaccaa gtccttgaac
     1141 agggaggtgt gtccagtttg tttcagaatc tcggggtgtc cgtaactccg atccaaagga
     1201 ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca tgtcatcatc ccgtatgaag
     1261 gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt taaggtggtg taccctgtgg
     1321 atgatcatca ctttaaggtg atcctgcact atggcacact ggtaatcgac ggggttacgc
     1381 cgaacatgat cgactatttc ggacggccgt atgaaggcat cgccgtgttc gacggcaaaa
     1441 agatcactgt aacagggacc ctgtggaacg gcaacaaaat tatcgacgag cgcctgatca
     1501 accccgacgg ctccctgctg ttccgagtaa ccatcaacgg agtgaccggc tggcggctgt
     1561 gcgaacgcat tctggcgggc tcgagcggcg cgatcgccat ggtgaggagc aggcaaatgt
     1621 gcaataccaa catgtctgta cctactgatg gtgctgtaac cacctcacag attccagctt
     1681 cggaacaaga gaccctggtt agaccaaagc cattgctttt gaagttatta aagtctgttg
     1741 gtgcacaaaa agacacttat actatgaaag aggttctttt ttatcttggc cagtatatta
     1801 tgactaaacg attatatgat gagaagcaac aacatattgt atattgttca aatgatcttc
     1861 taggagattt gtttggcgtg ccaagcttct ctgtgaaaga gcacaggaaa atatatacca
     1921 tgatctacag gaacttggta gtagtcaatc agcaggaatc atcggactca ggtacatctg
     1981 tgagtgagaa caggtgtcac cttgaaggtg ggagtgatca aaaggacctt gtacaagagc
     2041 ttcaggaaga gaaaccttca tcttcacatt tggtttctag accatctacc tcatctagaa
     2101 ggagagcaat tagtgagaca gaagaaaatt cagatgaatt atctggtgaa cgacaaagaa
     2161 aacgccacaa atctgatagt atttcccttt cctttgatga aagcctggct ctgtgtgtaa
     2221 taagggagat atgttgtgaa agaagcagta gcagtgaatc tacagggacg ccatcgaatc
     2281 cggatcttga tgctggtgta agtgaacatt caggtgattg gttggatcag gattcagttt
     2341 cagatcagtt tagtgtagaa tttgaagttg aatctctcga ctcagaagat tatagcctta
     2401 gtgaagaagg acaagaactc tcagatgaag atgatgaggt atatcaagtt actgtgtatc
     2461 aggcagggga gagtgataca gattcatttg aagaagatcc tgaaatttcc ttagctgact
     2521 attggaaatg cacttcatgc aatgaaatga atccccccct tccatcacat tgcaacagat
     2581 gttgggccct tcgtgagaat tggcttcctg aagataaagg gaaagataaa ggggaaatct
     2641 ctgagaaagc caaactggaa aactcaacac aagctgaaga gggctttgat gttcctgatt
     2701 gtaaaaaaac tatagtgaat gattccagag agtcatgtgt tgaggaaaat gatgataaaa
     2761 ttacacaagc ttcacaatca caagaaagtg aagactattc tcagccatca acttctagta
     2821 gcattattta tagcagccaa gaagatgtga aagagtttga aagggaagaa acccaagaca
     2881 aagaagagag tgtggaatct agtttgcccc ttaatgccat tgaaccttgt gtgatttgtc
     2941 aaggtcgacc taaaaatggt tgcattgtcc atggcaaaac aggacatctt atggcctgct
     3001 ttacatgtgc aaagaagcta aagaaaagga ataagccctg cccagtatgt agacaaccaa
     3061 ttcaaatgat tgtgctaact tatttccccg tttaaacgaa ttcgggctcg gtacccgggg
     3121 atcctctaga gtcgacctgc aggcatgcaa gctgatccgg ctgctaacaa agcccgaaag
     3181 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcggcc
     3241 gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     3301 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     3361 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     3421 ggagatgtgg gaggtttttt taagcaagta aaacctctac aaatgtggta aaatcgaatt
     3481 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
     3541 cggtatttca caccgcatac gcggatctgc gcagcaccat ggcctgaaat aacctctgaa
     3601 agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     3661 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3721 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3781 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3841 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3901 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     3961 ggaggcctag gcttttgcaa aaagcttaat taactgttga caattaatca tcggcatagt
     4021 atatcggcat agtataatac gacaaggtga ggaactaaac ccaggaggca gatcatgatt
     4081 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4141 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4201 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4261 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4321 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4381 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4441 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4501 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4561 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4621 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4681 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4741 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4801 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4861 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4921 cacgatggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt
     4981 gaatcgatag cgataaggat cctctttgcg cttgcgtttt cccttgtcca gatagcccag
     5041 tagctgacat tcatccgggg tcagcaccgt ttctgcggac tggctttcta cccggtatca
     5101 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5161 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5221 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5281 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5341 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5401 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5461 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5521 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5581 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5641 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5701 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5761 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatttcaaga agatcctttg
     5821 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5881 atgagattat caaaaaggat cttcacctag atccttttat agtccggaaa tacaggaacg
     5941 cacgctggat ggcccttcgc tgggatggtg aaaccatgaa aaatggcagc ttcagtggat
     6001 taagtggggg taatgtggcc tgtaccctct ggttgcatag gtattcatac ggttaaaatt
     6061 tatcaggcgc gattgcggca gtttttcggg tggtttgttg ccatttttac ctgtctgctg
     6121 ccgtgatcgc gctgaacgcg ttttagcggt gcgtacaatt aagggattat ggtaaatcca
     6181 cttactgtct gccctcgtag ccatcgagat aaaccgcagt actccggcca cgatgcgtcc
     6241 ggcgtagagg atcgagatct
//
