LOCUS       sequence-472431-        5413 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express NanoLuc(R)-CA2 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 5413)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5413)
  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            "Unpublished"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-CA2 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5413
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    42..163
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(79..98)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     133..152
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     200..218
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(1133..1645)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(1658..1677)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1659..1677)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1719..1738)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          1721..1853
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(1989..2192)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2018..2038)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2193..2572
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(3017..3034)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3151..3696)
                     /direction=LEFT
                     /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     complement(3231..3250)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3853..3901
                     /note="synthetic polyadenylation signal"
     CDS             complement(3965..4759)
                     /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(4076..4095)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4686..4705
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4791..4838)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(4853..5210)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4867..5002
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5019..5210
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5167..5187
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
       61 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
      121 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgctcgaagc
      181 ggccgcccca aggggttatg ctagttattg ctcagcggtg gcagcagcca actcagcttc
      241 ctttcgggct ttgttagcag ccggatcagc ttgcatgcct gcaggtcgac tctagaggat
      301 ccccgggtac cgagcccgaa ttcgtttaaa ctttgaagga agctttgatt tgcctgttct
      361 tcagtggctg agctgggcgc cagttgtcca ccatcagttc ttcgggttca ccctccccat
      421 tgaagttaag tttacggaat ttcaacacct gctcgctgct gacgctgatg ggttccttga
      481 gcacaatcca ggtcacacat tccagaagag gaggggtggt cagtgagcct gggtaggtcc
      541 agtaatccaa ggattcagga aggaggccac gaggatcgaa gttagtgaag tcagcactct
      601 tgccctttgt tttaatggaa tccagcacat caacaacttt ctgaaggccc ggtttagcgc
      661 tgccaacctt caaaaaaata cctagaacgg ccagtccatc aggttgctgc acagctttcc
      721 caaaatcccc atatttggtg ttccagtgaa ccaagtgaag ttctgcagca tatttctttt
      781 tatccacagt atgctctgaa ccttgtccat caagtgaacc ccagtgaaag tgaaactgaa
      841 tcaatctgta agtgccatcc aggggtcctc ccttgagcac tgctttgtcc tgagagtcat
      901 caaactccac gttgaaagca tgaccattgt tgaggatcct cagggaagtt gcttgatcat
      961 aggaaacaga caggggcttc agggaagggt catacttggc tgtatgagtg tcgatgtcaa
     1021 caggggactg gcgctctccc ttggcaatgg ggaagtcctt atgccagtgc tcaggtccgt
     1081 tgtgtttgcc gtacccccag tgatgggaca tggcgatcgc gccgctcgag cccgccagaa
     1141 tgcgttcgca cagccgccag ccggtcactc cgttgatggt tactcggaac agcagggagc
     1201 cgtcggggtt gatcaggcgc tcgtcgataa ttttgttgcc gttccacagg gtccctgtta
     1261 cagtgatctt tttgccgtcg aacacggcga tgccttcata cggccgtccg aaatagtcga
     1321 tcatgttcgg cgtaaccccg tcgattacca gtgtgccata gtgcaggatc accttaaagt
     1381 gatgatcatc cacagggtac accaccttaa aaattttttc gatctggccc atttggtcgc
     1441 cgctcagacc ttcatacggg atgatgacat ggatgtcgat cttcagccca ttttcaccgc
     1501 tcaggacaat cctttggatc ggagttacgg acaccccgag attctgaaac aaactggaca
     1561 cacctccctg ttcaaggact tggtccaggt tgtagccggc tgtctgtcgc cagtccccaa
     1621 cgaaatcttc gagtgtgaag accatggtga gcgctagccc tatagtgagt cgtattaata
     1681 ctctagcctt aagagctgta attgaactgg gagtggacac ctgtggagag aaaggcaaag
     1741 tggatgtcag taagaccaat aggtgcctat cagaaacgca agagtcttct ctgtctcgac
     1801 aagcccagtt tctattggtc tccttaaacc tgtcttgtaa ccttgatact tacctgccca
     1861 gtgcctcacg accaacttct gcagcttaag ttcgagactg ttgtgtcaga agcactgact
     1921 gcgttagcaa tttaactgtg ataaactacc gcaataaagc ttctagtgat ctgacggttc
     1981 actaaaccag ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc
     2041 aacggggcgg ggttattacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa
     2101 ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca
     2161 cgcccattgg tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag
     2221 atgtactgcc aagtaggaaa gtcccgtaag gtcatgtact gggcataatg ccaggcgggc
     2281 catttaccgt cattgacgtc aatagggggc ggacttggca tatgatacac ttgatgtact
     2341 gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt
     2401 ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt
     2461 cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga
     2521 actaatgacc ccgtaattga ttactattaa taactagtca ataatcaatg ccaacatggc
     2581 ggtcatattg gacatgagcc aatataaatg tacatattat gatatagata caacgtatgc
     2641 aatggccaat agccaatatt gatttatgct atataaccaa tgaataatat ggctaatggc
     2701 caatattgaa gatctcgatc ctctacgccg gacgcatcgt ggccggagta ctgcggttta
     2761 tctcgatggc tacgagggca gacagtaagt ggatttacca taatccctta attgtacgca
     2821 ccgctaaaac gcgtaatctc atgacggggt ggtgcgtaac ggcaaaagca ccgccggaca
     2881 tcagcgctag cggagtgtat actggcttac tatgttggca ctgatgaggg tgtcagtgaa
     2941 gtgcttcatg tggcaggaga aaaaaggctg caccggtgcg tcagcagaat atgtgataca
     3001 ggatatattc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag
     3061 cggaaatggc ttacgaacgg ggcggagatt tcctggaaga tgccaggaag atacttaaca
     3121 gggaagtgag agggccgcgg caaagccgtt tttccatagg ctccgccccc ctgacaagca
     3181 tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca
     3241 ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg
     3301 tgtcattccg ctgttatggc cgcgtttgtc tcattccacg cctgacactc agttccgggt
     3361 aggcagttcg ctccaagctg gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg
     3421 ccttatccgg taactatcgt cttgagtcca acccggaaag acatgcaaaa gcaccactgg
     3481 cagcagccac tggtaattga tttagaggag ttagtcttga agtcatgcgc cggttaaggc
     3541 taaactgaaa ggacaagttt tggtgactgc gctcctccaa gccagttacc tcggttcaaa
     3601 gagttggtag ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca
     3661 gagcaagaga ttacgcgcag accaaaacga tctcaagaag atcatcttat taatcagata
     3721 aaatatttct agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc
     3781 atacgatata agttgtaatt ctcatgtttg acagcttatc atcgcatgag cggatcctta
     3841 tcgctatcga ttcacacaaa aaaccaacac acagatgtaa tgaaaataaa gatattttat
     3901 tgcggccatc gtgatggcag gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc
     3961 ccgctcagaa gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg
     4021 cgataccgta aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat
     4081 cacgggtagc caacgctatg tcctgatagc ggtccgccac acccagccgg ccacagtcga
     4141 tgaatccaga aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatggg
     4201 tcacgacgag atcctcgccg tcgggcatgc gcgccttgag cctggcgaac agttcggctg
     4261 gcgcgagccc ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc
     4321 gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat
     4381 caagcgtatg cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa
     4441 ggtgagatga caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg
     4501 cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc agccacgata
     4561 gccgcgctgc ctcgtcctgc agttcattca gggcaccgga caggtcggtc ttgacaaaaa
     4621 gaaccgggcg cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct
     4681 gttgtgccca gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca
     4741 atccatcttg ttcaatcatg atctgcctcc tgggtttagt tcctcacctt gtcgtattat
     4801 actatgccga tatactatgc cgatgattaa ttgtcaacag ttaattaagc tttttgcaaa
     4861 agcctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg
     4921 cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac
     4981 tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat
     5041 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg
     5101 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
     5161 ttccacaccc taactgacac acattccaca gctggttctt tccgcctcag aaggtaccta
     5221 accaagttcc tctttcagag gttatttcag gccatggtgc tgcgcagatc cgcgtatgcg
     5281 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat tgtaagcgtt
     5341 aatattttgt taaaattcga ttttaccaca tttgtagagg ttttacttgc ttaaaaaaac
     5401 ctcccacatc tcc
//
