LOCUS       sequence-479819-        5350 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express NanoLuc-BCL2 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 5350)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5350)
  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            "Unpublished"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-BCL2 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5350
                     /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(2429..2447)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2484..2605
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2495..2514)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2549..2568
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2850..3207
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2850..3041
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2873..2893)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3058..3193
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3222..3269
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3301..4095
                     /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(3355..3374)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3965..3984
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4159..4207
                     /note="synthetic polyadenylation signal"
     rep_origin      4364..4909
                     /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     4810..4829
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5026..5043
                     /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 ggcgcacgct gggagaacag
     1621 ggtacgataa ccgggagata gtgatgaagt acatccatta taagctgtcg cagaggggct
     1681 acgagtggga tgcgggagat gtgggcgccg cgcccccggg ggccgccccc gcaccgggca
     1741 tcttctcctc ccagcccggg cacacgcccc atccagccgc atcccgggac ccggtcgcca
     1801 ggacctcgcc gctgcagacc ccggctgccc ccggcgccgc cgcggggcct gcgctcagcc
     1861 cggtgccacc tgtggtccac ctgaccctcc gccaggccgg cgacgacttc tcccgccgct
     1921 accgccgcga cttcgccgag atgtccagcc agctgcacct gacgcccttc accgcgcggg
     1981 gacgctttgc cacggtggtg gaggagctct tcagggacgg ggtgaactgg gggaggattg
     2041 tggccttctt tgagttcggt ggggtcatgt gtgtggagag cgtcaaccgg gagatgtcgc
     2101 ccctggtgga caacatcgcc ctgtggatga ctgagtacct gaaccggcac ctgcacacct
     2161 ggatccagga taacggaggc tgggatgcct ttgtggaact gtacggcccc agcatgcggc
     2221 ctctgtttga tttctcctgg ctgtctctga agactctgct cagtttggcc ctggtgggag
     2281 cttgcatcac cctgggtgcc tatctgggcc acaaggttta aacgaattcg ggctcggtac
     2341 ccggggatcc tctagagtcg acctgcaggc atgcaagctg atccggctgc taacaaagcc
     2401 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
     2461 gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     2521 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     2581 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     2641 tcagggggag atgtgggagg tttttttaag caagtaaaac ctctacaaat gtggtaaaat
     2701 cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca
     2761 tctgtgcggt atttcacacc gcatacgcgg atctgcgcag caccatggcc tgaaataacc
     2821 tctgaaagag gaacttggtt aggtaccttc tgaggcggaa agaaccagct gtggaatgtg
     2881 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2941 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3001 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3061 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3121 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3181 ttttttggag gcctaggctt ttgcaaaaag cttaattaac tgttgacaat taatcatcgg
     3241 catagtatat cggcatagta taatacgaca aggtgaggaa ctaaacccag gaggcagatc
     3301 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     3361 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     3421 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     3481 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     3541 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     3601 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     3661 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     3721 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     3781 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     3841 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     3901 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     3961 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4021 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4081 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     4141 tgccatcacg atggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt
     4201 ttgtgtgaat cgatagcgat aaggatccgc tcatgcgatg ataagctgtc aaacatgaga
     4261 attacaactt atatcgtatg gggctgactt caggtgctac atttgaagag ataaattgca
     4321 ctgaaatcta gaaatatttt atctgattaa taagatgatc ttcttgagat cgttttggtc
     4381 tgcgcgtaat ctcttgctct gaaaacgaaa aaaccgcctt gcagggcggt ttttcgaagg
     4441 ttctctgagc taccaactct ttgaaccgag gtaactggct tggaggagcg cagtcaccaa
     4501 aacttgtcct ttcagtttag ccttaaccgg cgcatgactt caagactaac tcctctaaat
     4561 caattaccag tggctgctgc cagtggtgct tttgcatgtc tttccgggtt ggactcaaga
     4621 cgatagttac cggataaggc gcagcggtcg gactgaacgg ggggttcgtg catacagtcc
     4681 agcttggagc gaactgccta cccggaactg agtgtcaggc gtggaatgag acaaacgcgg
     4741 ccataacagc ggaatgacac cggtaaaccg aaaggcagga acaggagagc gcacgaggga
     4801 gccgccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc accactgatt
     4861 tgagcgtcag atttcgtgat gcttgtcagg ggggcggagc ctatggaaaa acggctttgc
     4921 cgcggccctc tcacttccct gttaagtatc ttcctggcat cttccaggaa atctccgccc
     4981 cgttcgtaag ccatttccgc tcgccgcagt cgaacgaccg agcgtagcga gtcagtgagc
     5041 gaggaagcgg aatatatcct gtatcacata ttctgctgac gcaccggtgc agcctttttt
     5101 ctcctgccac atgaagcact tcactgacac cctcatcagt gccaacatag taagccagta
     5161 tacactccgc tagcgctgat gtccggcggt gcttttgccg ttacgcacca ccccgtcatg
     5221 agattacgcg ttttagcggt gcgtacaatt aagggattat ggtaaatcca cttactgtct
     5281 gccctcgtag ccatcgagat aaaccgcagt actccggcca cgatgcgtcc ggcgtagagg
     5341 atcgagatct
//
