LOCUS       sequence-479776-        5348 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express NanoLuc-BCL2L2 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 5348)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5348)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoLuc-BCL2L2 Vector
FEATURES             Location/Qualifiers
     source          1..5348
                     /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(2291..2309)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2346..2467
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2357..2376)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2411..2430
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2712..3069
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2712..2903
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2735..2755)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2920..3055
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3084..3131
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3163..3957
                     /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(3217..3236)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3827..3846
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4021..4069
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4308..4896)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4388..4407)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5012..5295
                     /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 ggcgacccca gcctcggccc
     1621 cagacacacg ggctctggtg gcagactttg taggttataa gctgaggcag aagggttatg
     1681 tctgtggagc tggccccggg gagggcccag cagctgaccc gctgcaccaa gccatgcggg
     1741 cagctggaga tgagttcgag acccgcttcc ggcgcacctt ctctgatctg gcggctcagc
     1801 tgcatgtgac cccaggctca gcccaacaac gcttcaccca ggtctccgat gaactttttc
     1861 aagggggccc caactggggc cgccttgtag ccttctttgt ctttggggct gcactgtgtg
     1921 ctgagagtgt caacaaggag atggaaccac tggtgggaca agtgcaggag tggatggtgg
     1981 cctacctgga gacgcggctg gctgactgga tccacagcag tgggggctgg gcggagttca
     2041 cagctctata cggtgacggg gctctggaag aggcgcggcg tctgcgggag ggtaactggg
     2101 catcagtgag gacagttctg acgggggccg tggcactggg ggccctggta actgtagggg
     2161 ccttttttgc tagcaaggtt taaacgaatt cgggctcggt acccggggat cctctagagt
     2221 cgacctgcag gcatgcaagc tgatccggct gctaacaaag cccgaaagga agctgagttg
     2281 gctgctgcca ccgctgagca ataactagca taaccccttg gggcggccgc ttcgagcaga
     2341 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     2401 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     2461 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga
     2521 ggttttttta agcaagtaaa acctctacaa atgtggtaaa atcgaatttt aacaaaatat
     2581 taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     2641 ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg
     2701 ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     2761 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     2821 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     2881 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     2941 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3001 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3061 ttttgcaaaa agcttaatta actgttgaca attaatcatc ggcatagtat atcggcatag
     3121 tataatacga caaggtgagg aactaaaccc aggaggcaga tcatgattga acaagatgga
     3181 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     3241 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     3301 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     3361 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     3421 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     3481 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     3541 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     3601 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     3661 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     3721 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     3781 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     3841 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     3901 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     3961 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgatggccgc
     4021 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagcg
     4081 ataaggatcc tctttgcgct tgcgttttcc cttgtccaga tagcccagta gctgacattc
     4141 atccggggtc agcaccgttt ctgcggactg gctttctacc cggtatcagc tcactcaaag
     4201 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4261 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4321 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4381 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4441 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4501 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4561 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4621 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4681 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4741 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4801 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4861 aagcagcaga ttacgcgcag aaaaaaagga tttcaagaag atcctttgat cttttctacg
     4921 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4981 aaaaggatct tcacctagat ccttttatag tccggaaata caggaacgca cgctggatgg
     5041 cccttcgctg ggatggtgaa accatgaaaa atggcagctt cagtggatta agtgggggta
     5101 atgtggcctg taccctctgg ttgcataggt attcatacgg ttaaaattta tcaggcgcga
     5161 ttgcggcagt ttttcgggtg gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc
     5221 tgaacgcgtt ttagcggtgc gtacaattaa gggattatgg taaatccact tactgtctgc
     5281 cctcgtagcc atcgagataa accgcagtac tccggccacg atgcgtccgg cgtagaggat
     5341 cgagatct
//
