LOCUS       sequence-479845-        5332 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express NanoLuc-BCL2L1 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 5332)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5332)
  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            "Unpublished"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-BCL2L1 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5332
                     /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(2411..2429)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2466..2587
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2477..2496)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2531..2550
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2832..3189
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2832..3023
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2855..2875)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3040..3175
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3204..3251
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3283..4077
                     /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(3337..3356)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3947..3966
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4141..4189
                     /note="synthetic polyadenylation signal"
     rep_origin      4346..4891
                     /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     4792..4811
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5008..5025
                     /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 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 gtctcagagc aaccgggagc
     1621 tggtggttga ctttctctcc tacaagcttt cccagaaagg atacagctgg agtcagttta
     1681 gtgatgtgga agagaacagg actgaggccc cagaagggac tgaatcggag atggagaccc
     1741 ccagtgccat caatggcaac ccatcctggc acctggcaga cagccccgcg gtgaatggag
     1801 ccactggcca cagcagcagt ttggatgccc gggaggtgat ccccatggca gcagtaaagc
     1861 aagcgctgag ggaggcaggc gacgagtttg aactgcggta ccggcgggca ttcagtgacc
     1921 tgacatccca gctccacatc accccaggga cagcatatca gagctttgaa caggtagtga
     1981 atgaactctt ccgggatggg gtaaactggg gtcgcattgt ggcctttttc tccttcggcg
     2041 gggcactgtg cgtggaaagc gtagacaagg agatgcaggt attggtgagt cggatcgcag
     2101 cttggatggc cacttacctg aatgaccacc tagagccttg gatccaggag aacggcggct
     2161 gggatacttt tgtggaactc tatgggaaca atgcagcagc cgagagccga aagggccagg
     2221 aacgcttcaa ccgctggttc ctgacgggca tgactgtggc cggcgtggtt ctgctgggct
     2281 cactcttcag tcggaaagtt taaacgaatt cgggctcggt acccggggat cctctagagt
     2341 cgacctgcag gcatgcaagc tgatccggct gctaacaaag cccgaaagga agctgagttg
     2401 gctgctgcca ccgctgagca ataactagca taaccccttg gggcggccgc ttcgagcaga
     2461 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     2521 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     2581 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga
     2641 ggttttttta agcaagtaaa acctctacaa atgtggtaaa atcgaatttt aacaaaatat
     2701 taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     2761 ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg
     2821 ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     2881 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     2941 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3001 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3061 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3121 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3181 ttttgcaaaa agcttaatta actgttgaca attaatcatc ggcatagtat atcggcatag
     3241 tataatacga caaggtgagg aactaaaccc aggaggcaga tcatgattga acaagatgga
     3301 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     3361 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     3421 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     3481 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     3541 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     3601 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     3661 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     3721 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     3781 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     3841 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     3901 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     3961 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4021 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4081 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgatggccgc
     4141 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagcg
     4201 ataaggatcc gctcatgcga tgataagctg tcaaacatga gaattacaac ttatatcgta
     4261 tggggctgac ttcaggtgct acatttgaag agataaattg cactgaaatc tagaaatatt
     4321 ttatctgatt aataagatga tcttcttgag atcgttttgg tctgcgcgta atctcttgct
     4381 ctgaaaacga aaaaaccgcc ttgcagggcg gtttttcgaa ggttctctga gctaccaact
     4441 ctttgaaccg aggtaactgg cttggaggag cgcagtcacc aaaacttgtc ctttcagttt
     4501 agccttaacc ggcgcatgac ttcaagacta actcctctaa atcaattacc agtggctgct
     4561 gccagtggtg cttttgcatg tctttccggg ttggactcaa gacgatagtt accggataag
     4621 gcgcagcggt cggactgaac ggggggttcg tgcatacagt ccagcttgga gcgaactgcc
     4681 tacccggaac tgagtgtcag gcgtggaatg agacaaacgc ggccataaca gcggaatgac
     4741 accggtaaac cgaaaggcag gaacaggaga gcgcacgagg gagccgccag ggggaaacgc
     4801 ctggtatctt tatagtcctg tcgggtttcg ccaccactga tttgagcgtc agatttcgtg
     4861 atgcttgtca ggggggcgga gcctatggaa aaacggcttt gccgcggccc tctcacttcc
     4921 ctgttaagta tcttcctggc atcttccagg aaatctccgc cccgttcgta agccatttcc
     4981 gctcgccgca gtcgaacgac cgagcgtagc gagtcagtga gcgaggaagc ggaatatatc
     5041 ctgtatcaca tattctgctg acgcaccggt gcagcctttt ttctcctgcc acatgaagca
     5101 cttcactgac accctcatca gtgccaacat agtaagccag tatacactcc gctagcgctg
     5161 atgtccggcg gtgcttttgc cgttacgcac caccccgtca tgagattacg cgttttagcg
     5221 gtgcgtacaa ttaagggatt atggtaaatc cacttactgt ctgccctcgt agccatcgag
     5281 ataaaccgca gtactccggc cacgatgcgt ccggcgtaga ggatcgagat ct
//
