LOCUS       sequence-474739-        5129 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express NanoLuc(R)-CECR2(BD) 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 5129)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5129)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoLuc-CECR2(BD) Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5129
                     /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(2072..2090)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2127..2248
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2138..2157)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2192..2211
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2493..2850
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2493..2684
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2516..2536)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2701..2836
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2865..2912
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2944..3738
                     /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="MIEQDGLHAGSPAAWVVRLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2998..3017)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3608..3627
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3802..3850
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4089..4677)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4169..4188)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4793..5076
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
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 gagagaggaa aaaaagacta
     1621 aagacctctt tgagttggat gatgatttca ctgctatgta taaagttcta gacgtggtaa
     1681 aggctcacaa ggattcctgg cccttcttgg aacctgtgga tgaatcttat gcccctaact
     1741 attatcagat tattaaggcc cccatggata tttccagcat ggagaagaaa ctgaatggag
     1801 gtttatactg taccaaggag gaatttgtaa atgacatgaa gaccatgttc aggaattgtc
     1861 gaaagtataa tggggaaagt agtgagtata ccaagatgtc tgataattta gagaggtgtt
     1921 tccatcgggc aatgatgaaa cattttcctg gagaagatgt ttaaacgaat tcgggctcgg
     1981 tacccgggga tcctctagag tcgacctgca ggcatgcaag ctgatccggc tgctaacaaa
     2041 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt
     2101 ggggcggccg cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac
     2161 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     2221 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     2281 ggttcagggg gagatgtggg aggttttttt aagcaagtaa aacctctaca aatgtggtaa
     2341 aatcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac
     2401 gcatctgtgc ggtatttcac accgcatacg cggatctgcg cagcaccatg gcctgaaata
     2461 acctctgaaa gaggaacttg gttaggtacc ttctgaggcg gaaagaacca gctgtggaat
     2521 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2581 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     2641 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     2701 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     2761 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     2821 ggcttttttg gaggcctagg cttttgcaaa aagcttaatt aactgttgac aattaatcat
     2881 cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc caggaggcag
     2941 atcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggtgaggcta
     3001 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3061 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3121 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3181 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3241 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3301 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3361 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3421 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     3481 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     3541 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     3601 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     3661 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     3721 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     3781 acctgccatc acgatggccg caataaaata tctttatttt cattacatct gtgtgttggt
     3841 tttttgtgtg aatcgatagc gataaggatc ctctttgcgc ttgcgttttc ccttgtccag
     3901 atagcccagt agctgacatt catccggggt cagcaccgtt tctgcggact ggctttctac
     3961 ccggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4021 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4081 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4141 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4201 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4261 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4321 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4381 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4441 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4501 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     4561 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4621 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atttcaagaa
     4681 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4741 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttata gtccggaaat
     4801 acaggaacgc acgctggatg gcccttcgct gggatggtga aaccatgaaa aatggcagct
     4861 tcagtggatt aagtgggggt aatgtggcct gtaccctctg gttgcatagg tattcatacg
     4921 gttaaaattt atcaggcgcg attgcggcag tttttcgggt ggtttgttgc catttttacc
     4981 tgtctgctgc cgtgatcgcg ctgaacgcgt tttagcggtg cgtacaatta agggattatg
     5041 gtaaatccac ttactgtctg ccctcgtagc catcgagata aaccgcagta ctccggccac
     5101 gatgcgtccg gcgtagagga tcgagatct
//
