LOCUS       sequence-473343-        6380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express NanoLuc(R)-FYN 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 6380)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6380)
  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            "Unpublished"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-FYN Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6380
                     /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(3323..3341)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3378..3499
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3389..3408)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3443..3462
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3744..4101
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3744..3935
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3767..3787)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3952..4087
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4116..4163
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4195..4989
                     /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(4249..4268)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4859..4878
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5053..5101
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5340..5928)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5420..5439)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6044..6327
                     /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 gggctgtgtg caatgtaagg
     1621 ataaagaagc aacaaaactg acggaggaga gggacggcag cctgaaccag agctctgggt
     1681 accgctatgg cacagacccc acccctcagc actaccccag cttcggtgtg acctccatcc
     1741 ccaactacaa caacttccac gcagccgggg gccaaggact caccgtcttt ggaggtgtga
     1801 actcttcgtc tcatacgggg accttgcgta cgagaggagg aacaggagtg acactctttg
     1861 tggcccttta tgactatgaa gcacggacag aagatgacct gagttttcac aaaggagaaa
     1921 aatttcaaat attgaacagc tcggaaggag attggtggga agcccgctcc ttgacaactg
     1981 gagagacagg ttacattccc agcaattatg tggctccagt tgactctatc caggcagaag
     2041 agtggtactt tggaaaactt ggccgaaaag atgctgagcg acagctattg tcctttggaa
     2101 acccaagagg tacctttctt atccgcgaga gtgaaaccac caaaggtgcc tattcacttt
     2161 ctatccgtga ttgggatgat atgaaaggag accatgtcaa acattataaa attcgcaaac
     2221 ttgacaatgg tggatactac attaccaccc gggcccagtt tgaaacactt cagcagcttg
     2281 tacaacatta ctcagagaga gctgcaggtc tctgctgccg cctagtagtt ccctgtcaca
     2341 aagggatgcc aaggcttacc gatctgtctg tcaaaaccaa agatgtctgg gaaatccctc
     2401 gagaatccct gcagttgatc aagagactgg gaaatgggca gtttggggaa gtatggatgg
     2461 gtacctggaa tggaaacaca aaagtagcca taaagactct taaaccaggc acaatgtccc
     2521 ccgaatcatt ccttgaggaa gcgcagatca tgaagaagct gaagcacgac aagctggtcc
     2581 agctctatgc agtggtgtct gaggagccca tctacatcgt caccgagtat atgaacaaag
     2641 gaagtttact ggatttctta aaagatggag aaggaagagc tctgaaatta ccaaatcttg
     2701 tggacatggc agcacaggtg gctgcaggaa tggcttacat cgagcgcatg aattatatcc
     2761 atagagatct gcgatcagca aacattctag tggggaatgg actcatatgc aagattgctg
     2821 acttcggatt ggcccgattg atagaagaca atgagtacac agcaagacaa ggtgcaaagt
     2881 tccccatcaa gtggacggcc cccgaggcag ccctgtacgg gaggttcaca atcaagtctg
     2941 acgtgtggtc ttttggaatc ttactcacag agctggtcac caaaggaaga gtgccatacc
     3001 caggcatgaa caaccgggag gtgctggagc aggtggagcg aggctacagg atgccctgcc
     3061 cgcaggactg ccccatctct ctgcatgagc tcatgatcca ctgctggaaa aaggaccctg
     3121 aagaacgccc cacttttgag tacttgcaga gcttcctgga agactacttt accgcgacag
     3181 agccccagta ccaacctggt gaaaacctgg tttaaacgaa ttcgggctcg gtacccgggg
     3241 atcctctaga gtcgacctgc aggcatgcaa gctgatccgg ctgctaacaa agcccgaaag
     3301 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcggcc
     3361 gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     3421 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     3481 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     3541 ggagatgtgg gaggtttttt taagcaagta aaacctctac aaatgtggta aaatcgaatt
     3601 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
     3661 cggtatttca caccgcatac gcggatctgc gcagcaccat ggcctgaaat aacctctgaa
     3721 agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     3781 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3841 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3901 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3961 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4021 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     4081 ggaggcctag gcttttgcaa aaagcttaat taactgttga caattaatca tcggcatagt
     4141 atatcggcat agtataatac gacaaggtga ggaactaaac ccaggaggca gatcatgatt
     4201 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4261 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4321 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4381 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4441 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4501 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4561 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4621 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4681 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4741 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4801 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4861 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4921 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4981 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5041 cacgatggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt
     5101 gaatcgatag cgataaggat cctctttgcg cttgcgtttt cccttgtcca gatagcccag
     5161 tagctgacat tcatccgggg tcagcaccgt ttctgcggac tggctttcta cccggtatca
     5221 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5281 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5341 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5401 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5461 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5521 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5581 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5641 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5701 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5761 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5821 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5881 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatttcaaga agatcctttg
     5941 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6001 atgagattat caaaaaggat cttcacctag atccttttat agtccggaaa tacaggaacg
     6061 cacgctggat ggcccttcgc tgggatggtg aaaccatgaa aaatggcagc ttcagtggat
     6121 taagtggggg taatgtggcc tgtaccctct ggttgcatag gtattcatac ggttaaaatt
     6181 tatcaggcgc gattgcggca gtttttcggg tggtttgttg ccatttttac ctgtctgctg
     6241 ccgtgatcgc gctgaacgcg ttttagcggt gcgtacaatt aagggattat ggtaaatcca
     6301 cttactgtct gccctcgtag ccatcgagat aaaccgcagt actccggcca cgatgcgtcc
     6361 ggcgtagagg atcgagatct
//
