LOCUS       sequence-479838-        5324 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express BCL2-NanoLuc 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 5324)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5324)
  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            "Unpublished"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: BCL2-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5324
                     /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             1803..2315
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2403..2421)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2458..2579
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2469..2488)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2523..2542
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2824..3181
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2824..3015
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2847..2867)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3032..3167
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3196..3243
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3275..4069
                     /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(3329..3348)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3939..3958
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4133..4181
                     /note="synthetic polyadenylation signal"
     rep_origin      4338..4883
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     4784..4803
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5000..5017
                     /label=L4440
                     /note="L4440 vector, forward primer"
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 ggctagcgat cgccatggcg cacgctggga
     1081 gaacagggta cgataaccgg gagatagtga tgaagtacat ccattataag ctgtcgcaga
     1141 ggggctacga gtgggatgcg ggagatgtgg gcgccgcgcc cccgggggcc gcccccgcac
     1201 cgggcatctt ctcctcccag cccgggcaca cgccccatcc agccgcatcc cgggacccgg
     1261 tcgccaggac ctcgccgctg cagaccccgg ctgcccccgg cgccgccgcg gggcctgcgc
     1321 tcagcccggt gccacctgtg gtccacctga ccctccgcca ggccggcgac gacttctccc
     1381 gccgctaccg ccgcgacttc gccgagatgt ccagccagct gcacctgacg cccttcaccg
     1441 cgcggggacg ctttgccacg gtggtggagg agctcttcag ggacggggtg aactggggga
     1501 ggattgtggc cttctttgag ttcggtgggg tcatgtgtgt ggagagcgtc aaccgggaga
     1561 tgtcgcccct ggtggacaac atcgccctgt ggatgactga gtacctgaac cggcacctgc
     1621 acacctggat ccaggataac ggaggctggg atgcctttgt ggaactgtac ggccccagca
     1681 tgcggcctct gtttgatttc tcctggctgt ctctgaagac tctgctcagt ttggccctgg
     1741 tgggagcttg catcaccctg ggtgcctatc tgggccacaa ggtttctctc ggctcgagcg
     1801 gcgtcttcac actcgaagat ttcgttgggg actggcgaca gacagccggc tacaacctgg
     1861 accaagtcct tgaacaggga ggtgtgtcca gtttgtttca gaatctcggg gtgtccgtaa
     1921 ctccgatcca aaggattgtc ctgagcggtg aaaatgggct gaagatcgac atccatgtca
     1981 tcatcccgta tgaaggtctg agcggcgacc aaatgggcca gatcgaaaaa atttttaagg
     2041 tggtgtaccc tgtggatgat catcacttta aggtgatcct gcactatggc acactggtaa
     2101 tcgacggggt tacgccgaac atgatcgact atttcggacg gccgtatgaa ggcatcgccg
     2161 tgttcgacgg caaaaagatc actgtaacag ggaccctgtg gaacggcaac aaaattatcg
     2221 acgagcgcct gatcaacccc gacggctccc tgctgttccg agtaaccatc aacggagtga
     2281 ccggctggcg gctgtgcgaa cgcattctgg cgtaaggccg cgactctaga gtcgacctgc
     2341 aggcatgcaa gctgatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     2401 caccgctgag caataactag cataacccct tggggcggcc gcttcgagca gacatgataa
     2461 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     2521 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     2581 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt
     2641 taagcaagta aaacctctac aaatgtggta aaatcgaatt ttaacaaaat attaacgctt
     2701 acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     2761 gcggatctgc gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttaggtac
     2821 cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     2881 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     2941 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3001 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3061 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3121 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3181 aaagcttaat taactgttga caattaatca tcggcatagt atatcggcat agtataatac
     3241 gacaaggtga ggaactaaac ccaggaggca gatcatgatt gaacaagatg gattgcacgc
     3301 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     3361 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     3421 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     3481 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     3541 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     3601 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     3661 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     3721 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     3781 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     3841 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     3901 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     3961 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4021 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     4081 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgatggcc gcaataaaat
     4141 atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag cgataaggat
     4201 ccgctcatgc gatgataagc tgtcaaacat gagaattaca acttatatcg tatggggctg
     4261 acttcaggtg ctacatttga agagataaat tgcactgaaa tctagaaata ttttatctga
     4321 ttaataagat gatcttcttg agatcgtttt ggtctgcgcg taatctcttg ctctgaaaac
     4381 gaaaaaaccg ccttgcaggg cggtttttcg aaggttctct gagctaccaa ctctttgaac
     4441 cgaggtaact ggcttggagg agcgcagtca ccaaaacttg tcctttcagt ttagccttaa
     4501 ccggcgcatg acttcaagac taactcctct aaatcaatta ccagtggctg ctgccagtgg
     4561 tgcttttgca tgtctttccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4621 gtcggactga acggggggtt cgtgcataca gtccagcttg gagcgaactg cctacccgga
     4681 actgagtgtc aggcgtggaa tgagacaaac gcggccataa cagcggaatg acaccggtaa
     4741 accgaaaggc aggaacagga gagcgcacga gggagccgcc agggggaaac gcctggtatc
     4801 tttatagtcc tgtcgggttt cgccaccact gatttgagcg tcagatttcg tgatgcttgt
     4861 caggggggcg gagcctatgg aaaaacggct ttgccgcggc cctctcactt ccctgttaag
     4921 tatcttcctg gcatcttcca ggaaatctcc gccccgttcg taagccattt ccgctcgccg
     4981 cagtcgaacg accgagcgta gcgagtcagt gagcgaggaa gcggaatata tcctgtatca
     5041 catattctgc tgacgcaccg gtgcagcctt ttttctcctg ccacatgaag cacttcactg
     5101 acaccctcat cagtgccaac atagtaagcc agtatacact ccgctagcgc tgatgtccgg
     5161 cggtgctttt gccgttacgc accaccccgt catgagatta cgcgttttag cggtgcgtac
     5221 aattaaggga ttatggtaaa tccacttact gtctgccctc gtagccatcg agataaaccg
     5281 cagtactccg gccacgatgc gtccggcgta gaggatcgag atct
//
