LOCUS       sequence-479839-        5306 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express BCL2L1-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 5306)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5306)
  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            "Unpublished"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: BCL2L1-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5306
                     /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             1785..2297
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2385..2403)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2440..2561
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2451..2470)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2505..2524
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2806..3163
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2806..2997
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2829..2849)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3014..3149
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3178..3225
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3257..4051
                     /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(3311..3330)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3921..3940
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4115..4163
                     /note="synthetic polyadenylation signal"
     rep_origin      4320..4865
                     /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     4766..4785
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4982..4999
                     /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 cgccatgtct cagagcaacc
     1081 gggagctggt ggttgacttt ctctcctaca agctttccca gaaaggatac agctggagtc
     1141 agtttagtga tgtggaagag aacaggactg aggccccaga agggactgaa tcggagatgg
     1201 agacccccag tgccatcaat ggcaacccat cctggcacct ggcagacagc cccgcggtga
     1261 atggagccac tggccacagc agcagtttgg atgcccggga ggtgatcccc atggcagcag
     1321 taaagcaagc gctgagggag gcaggcgacg agtttgaact gcggtaccgg cgggcattca
     1381 gtgacctgac atcccagctc cacatcaccc cagggacagc atatcagagc tttgaacagg
     1441 tagtgaatga actcttccgg gatggggtaa actggggtcg cattgtggcc tttttctcct
     1501 tcggcggggc actgtgcgtg gaaagcgtag acaaggagat gcaggtattg gtgagtcgga
     1561 tcgcagcttg gatggccact tacctgaatg accacctaga gccttggatc caggagaacg
     1621 gcggctggga tacttttgtg gaactctatg ggaacaatgc agcagccgag agccgaaagg
     1681 gccaggaacg cttcaaccgc tggttcctga cgggcatgac tgtggccggc gtggttctgc
     1741 tgggctcact cttcagtcgg aaagtttctc tcggctcgag cggcgtcttc acactcgaag
     1801 atttcgttgg ggactggcga cagacagccg gctacaacct ggaccaagtc cttgaacagg
     1861 gaggtgtgtc cagtttgttt cagaatctcg gggtgtccgt aactccgatc caaaggattg
     1921 tcctgagcgg tgaaaatggg ctgaagatcg acatccatgt catcatcccg tatgaaggtc
     1981 tgagcggcga ccaaatgggc cagatcgaaa aaatttttaa ggtggtgtac cctgtggatg
     2041 atcatcactt taaggtgatc ctgcactatg gcacactggt aatcgacggg gttacgccga
     2101 acatgatcga ctatttcgga cggccgtatg aaggcatcgc cgtgttcgac ggcaaaaaga
     2161 tcactgtaac agggaccctg tggaacggca acaaaattat cgacgagcgc ctgatcaacc
     2221 ccgacggctc cctgctgttc cgagtaacca tcaacggagt gaccggctgg cggctgtgcg
     2281 aacgcattct ggcgtaaggc cgcgactcta gagtcgacct gcaggcatgc aagctgatcc
     2341 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     2401 agcataaccc cttggggcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg
     2461 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     2521 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     2581 attttatgtt tcaggttcag ggggagatgt gggaggtttt tttaagcaag taaaacctct
     2641 acaaatgtgg taaaatcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt
     2701 attttctcct tacgcatctg tgcggtattt cacaccgcat acgcggatct gcgcagcacc
     2761 atggcctgaa ataacctctg aaagaggaac ttggttaggt accttctgag gcggaaagaa
     2821 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     2881 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     2941 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3001 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3061 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     3121 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctta attaactgtt
     3181 gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa
     3241 acccaggagg cagatcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     3301 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     3361 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     3421 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     3481 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     3541 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     3601 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     3661 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     3721 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     3781 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     3841 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     3901 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     3961 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     4021 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     4081 caagcgacgc ccaacctgcc atcacgatgg ccgcaataaa atatctttat tttcattaca
     4141 tctgtgtgtt ggttttttgt gtgaatcgat agcgataagg atccgctcat gcgatgataa
     4201 gctgtcaaac atgagaatta caacttatat cgtatggggc tgacttcagg tgctacattt
     4261 gaagagataa attgcactga aatctagaaa tattttatct gattaataag atgatcttct
     4321 tgagatcgtt ttggtctgcg cgtaatctct tgctctgaaa acgaaaaaac cgccttgcag
     4381 ggcggttttt cgaaggttct ctgagctacc aactctttga accgaggtaa ctggcttgga
     4441 ggagcgcagt caccaaaact tgtcctttca gtttagcctt aaccggcgca tgacttcaag
     4501 actaactcct ctaaatcaat taccagtggc tgctgccagt ggtgcttttg catgtctttc
     4561 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcggact gaacgggggg
     4621 ttcgtgcata cagtccagct tggagcgaac tgcctacccg gaactgagtg tcaggcgtgg
     4681 aatgagacaa acgcggccat aacagcggaa tgacaccggt aaaccgaaag gcaggaacag
     4741 gagagcgcac gagggagccg ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4801 ttcgccacca ctgatttgag cgtcagattt cgtgatgctt gtcagggggg cggagcctat
     4861 ggaaaaacgg ctttgccgcg gccctctcac ttccctgtta agtatcttcc tggcatcttc
     4921 caggaaatct ccgccccgtt cgtaagccat ttccgctcgc cgcagtcgaa cgaccgagcg
     4981 tagcgagtca gtgagcgagg aagcggaata tatcctgtat cacatattct gctgacgcac
     5041 cggtgcagcc ttttttctcc tgccacatga agcacttcac tgacaccctc atcagtgcca
     5101 acatagtaag ccagtataca ctccgctagc gctgatgtcc ggcggtgctt ttgccgttac
     5161 gcaccacccc gtcatgagat tacgcgtttt agcggtgcgt acaattaagg gattatggta
     5221 aatccactta ctgtctgccc tcgtagccat cgagataaac cgcagtactc cggccacgat
     5281 gcgtccggcg tagaggatcg agatct
//
