LOCUS       sequence-481547-        6162 bp    DNA     circular SYN 30-JUL-2025
DEFINITION  Express NanoLuc-BCL2L11 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 6162)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6162)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 30, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoLuc-BCL2L11 Vector
FEATURES             Location/Qualifiers
     source          1..6162
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    17..98
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     42..61
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(916..1428)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     promoter        complement(1467..1485)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1468..1485)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        1605..1908
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2063..2083
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        2116..2163
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2195..2854
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2261..2280)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(3026..3614)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3106..3125)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3740..3788
                     /note="synthetic polyadenylation signal"
     CDS             complement(3812..4606)
                     /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(3923..3942)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4533..4552
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4984..6162)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4993..5931
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(5016..5036)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 aagacgcgtg acgtcggcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
       61 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
      121 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
      181 aatgtggtat ggctgattat gatccgcggc cgcaatacgt cgacgttatc agctgacttc
      241 gtacgagagc ctaggtgttt ctagaggatc cccgggtacc gagcccgaat tcgtttaaac
      301 atgcattctc cacaccaggc ggacaatgta acgtaacagt cgtaagataa ccattcgtgg
      361 gtggtcttcg gctgcttggt aattattcaa aaataccctc cttgcatagt aagcgttaaa
      421 ctcgtctcca atacgccgca actcttgggc gatccatatc tctgggcgca tatctgcagg
      481 ttcagcctgc ctcatggaag ccattgcact gagatagtgg ttgaaggcct ggcaaggagg
      541 acttggggtt tgtgttgatt tgtcacaact catgggtgct gggctcctgt ctgtgtcaaa
      601 agagaaatac ccactggagg atcgagacag cagggaggat cttctcataa agatgaaaag
      661 cggggatctg gtagcaaaag ggccagggct ggcaggtggg gccagcgggc cctgagggct
      721 gccgtggggg cagctgtccc cttcacctcc gtgattgcct tcaggattac cttgtggctc
      781 tgtctgtagg gaggtagggg ccccaggtct gagctgggga ggcctctccg caggctgcaa
      841 ttgtctacct tctcggtcac actcagaact tacatcagaa ggttgctttg ccatggcgat
      901 cgcgccgctc gagcccgcca gaatgcgttc gcacagccgc cagccggtca ctccgttgat
      961 ggttactcgg aacagcaggg agccgtcggg gttgatcagg cgctcgtcga taattttgtt
     1021 gccgttccac agggtccctg ttacagtgat ctttttgccg tcgaacacgg cgatgccttc
     1081 atacggccgt ccgaaatagt cgatcatgtt cggcgtaacc ccgtcgatta ccagtgtgcc
     1141 atagtgcagg atcaccttaa agtgatgatc atccacaggg tacaccacct taaaaatttt
     1201 ttcgatctgg cccatttggt cgccgctcag accttcatac gggatgatga catggatgtc
     1261 gatcttcagc ccattttcac cgctcaggac aatcctttgg atcggagtta cggacacccc
     1321 gagattctga aacaaactgg acacacctcc ctgttcaagg acttggtcca ggttgtagcc
     1381 ggctgtctgt cgccagtccc caacgaaatc ttcgagtgtg aagaccatgg tgagcgctag
     1441 ctgcaaaaag aacaagtagc ttgtattcta tagtgtcacc taaatgggcg atctgacggt
     1501 tcactaaacc agctctgctt atataggcct ccctggcgcg ccactagtta ttaatagtaa
     1561 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     1621 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     1681 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     1741 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtcc gccccctatt
     1801 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttacgggac
     1861 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     1921 tggcagtaca ccaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     1981 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     2041 cgtaataacc ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtcaaa
     2101 tcgaattcta atactgttga caattaatca tcggcatagt atatcggcat agtataatac
     2161 gacaaggtga ggaactaaac ccaggaggca gatcatggag aaaaaaatca ctggatatac
     2221 caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc
     2281 tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa
     2341 gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaatgc
     2401 tcatccggaa ttccgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca
     2461 cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct ggagtgaata
     2521 ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga
     2581 aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtct cagccaatcc
     2641 ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc
     2701 cgttttcact atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat
     2761 tcaggttcat catgccgtct gtgatggctt ccatgtcggc agaatgctta atgaattaca
     2821 acagtactgc gatgagtggc agggcggggc gtaagcggga ctctggggtt cgaaatgacc
     2881 gaccaagcga cgcccaaccg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     2941 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3001 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3061 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3121 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3181 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3241 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3301 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3361 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3421 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     3481 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     3541 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     3601 aaaaaggatt tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     3661 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     3721 ttttatagtc cggaatcgtc acacaaaaaa ccaacacaca gatgtaatga aaataaagat
     3781 attttattgc ggccatcgtg atggctagag ttcagaagaa ctcgtcaaga aggcgataga
     3841 aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag cggtcagccc
     3901 attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc tgatagcggt
     3961 ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt tccaccatga
     4021 tattcggcaa gcaggcatcg ccgtgggtca cgacgagatc ctcgccgtcg ggcatgcgcg
     4081 ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg tccagatcat
     4141 cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt
     4201 ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca
     4261 tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc cccggcactt
     4321 cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag
     4381 gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcctgcagt tcattcaggg
     4441 caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca
     4501 cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat agcctctcca
     4561 cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatggtg gcatcgattc
     4621 ccggtcaaca tccagtacac ctccggcaca ccggcttccc caagggcgcc acgctctccc
     4681 accacaacat cctcaacaac ggctactggg tgggccgcac ggtcggctac acggagcagg
     4741 accgggtgtg tctgccggtg cccttctacc actgcttcgg catggtcatg ggcaacttgg
     4801 gcgccacctc ccacgccgcc tgcatcgtca tcccggcccc gtccttcgag ccggcggcga
     4861 cgctggaggc ggtgcagcgg gagaggtgca cgtccctgta cggcgtcccc acgaagccga
     4921 attccaccac actggactag atgcatgctc gacagtgtga tggatatctg cagaattcgg
     4981 ctttcacgac acctgaaatg gaagaaaaaa actttgaacc actgtctgag gcttgagaat
     5041 gaaccaagat ccaaactcaa aaagggcaaa ttccaaggag aattacatca agtgccaagc
     5101 tggcctaact tcagtctcca cccactcagt gtggggaaac tccatcgcat aaaacccctc
     5161 cccccaacct aaagacgacg tactccaaaa gctcgagaac taatcgaggt gcctggacgg
     5221 cgcccggtac tccgtggagt cacatgaagc gacggctgag gacggaaagg cccttttcct
     5281 ttgtgtgggt gactcacccg cccgctctcc cgagcgccgc gtcctccatt ttgagctgcc
     5341 tgcagcaggg ccgggaagcg gccatctttc cgctcacgca actggtgccg accgggccag
     5401 ccttgccgcc cagggcgggg cgatacacgg cggcgcgagg ccaggcacca gagcaggccg
     5461 gccagcttga gactaccccc gtccgattct cggtggccgc gctcgcaggc cccgcctcgc
     5521 cgaacatgtg cgctgggacg cacgggcccc gtcgccgccc gcggccccaa aaaccgaaat
     5581 accagtgtgc agatcttggc ccgcatttac aagactatct tgccagaaaa aaagcgtcgc
     5641 agcaggtcat caaaaatttt aaatggctag agacttatcg aaagcagcga gacaggcgcg
     5701 aaggtgccac cagattcgca cgcggcggcc ccagcgccca ggccaggcct caactcaagc
     5761 acgaggcgaa ggggctcctt aagcgcaagg cctcgaactc tcccacccac ttccaacccg
     5821 aagctcggga tcaagaatca cgtactgcag ccaggtggaa gtaattcaag gcacgcaagg
     5881 gccataaccc gtaaagaggc caggcccgcg ggaaccacac acggcactta cctgtgttct
     5941 ggcggcaaac ccgttgcgaa aaagaacgtt cacggcgact actgcactta tatacggttc
     6001 tcccccaccc tcgggaaaaa ggcggagcca gtacacgaca tcactttccc agtttacccc
     6061 gcgccacctt ctctaggcac cggttcaatt gccgacccct ccccccaact tctcggggac
     6121 tgtgggcgat gtgcgctctg cccactgacg ggcaccggag cc
//
