LOCUS       sequence-479854-        6648 bp    DNA     circular SYN 05-AUG-2025
DEFINITION  Express Syk-NanoLuc 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 6648)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6648)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoBRET Assay Vector, Syk-NanoLuc
FEATURES             Location/Qualifiers
     source          1..6648
                     /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             2991..3503
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(3591..3609)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3646..3767
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3657..3676)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3711..3730
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4012..4369
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4012..4203
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4035..4055)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4220..4355
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4384..4431
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4463..5257
                     /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(4517..4536)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5127..5146
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5321..5369
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5608..6196)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5688..5707)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6312..6595
                     /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 ggctagcgat cgccatggcc agcagcggca
     1081 tggctgacag cgccaaccac ctgcccttct ttttcggcaa catcacccgg gaggaggcag
     1141 aagattacct ggtccagggg ggcatgagtg atgggcttta tttgctgcgc cagagccgca
     1201 actacctggg tggcttcgcc ctgtccgtgg cccacgggag gaaggcacac cactacacca
     1261 tcgagcggga gctgaatggc acctacgcca tcgccggtgg caggacccat gccagccccg
     1321 ccgacctctg ccactaccac tcccaggagt ctgatggcct ggtctgcctc ctcaagaagc
     1381 ccttcaaccg gccccaaggg gtgcagccca agactgggcc ctttgaggat ttgaaggaaa
     1441 acctcatcag ggaatatgtg aagcagacat ggaacctgca gggtcaggct ctggagcagg
     1501 ccatcatcag tcagaagcct cagctggaga agctgatcgc taccacagcc catgaaaaaa
     1561 tgccttggtt ccatggaaaa atctctcggg aagaatctga gcaaattgtc ctgataggat
     1621 caaagacaaa tggaaagttc ctgatccgag ccagagacaa caacggctcc tacgccctgt
     1681 gcctgctgca cgaagggaag gtgctgcact atcgcatcga caaagacaag acagggaagc
     1741 tctccatccc cgagggaaag aagttcgaca cgctctggca gctagtcgag cattattctt
     1801 ataaagcaga tggtttgtta agagttctta ctgtcccatg tcaaaaaatc ggcacacagg
     1861 gaaatgttaa ttttggaggc cgtccacaac ttccaggttc ccatcctgcg acttggtcag
     1921 cgggtggaat aatctcaaga atcaaatcat actccttccc aaagcctggc cacagaaagt
     1981 cctcccctgc ccaagggaac cggcaagaga gtactgtgtc attcaatccg tatgagccag
     2041 aacttgcacc ctgggctgca gacaaaggcc cccagagaga agccctaccc atggacacag
     2101 aggtgtacga gagcccctac gcggaccccg aggagatcag gcccaaggag gtttacctgg
     2161 accgaaagct gctgacgctg gaagacaaag aactgggctc tggtaatttt ggaactgtga
     2221 aaaagggcta ctaccaaatg aaaaaagttg tgaaaaccgt ggctgtgaaa atactgaaaa
     2281 acgaggccaa tgaccccgct cttaaagatg agttattagc agaagcaaat gtcatgcagc
     2341 agctggacaa cccgtacatc gtgcggatga tcgggatatg cgaggccgag tcctggatgc
     2401 tggttatgga gatggcagaa cttggtcccc tcaataagta tttgcagcag aacagacatg
     2461 tcaaggataa gaacatcata gaactggttc atcaggtttc catgggcatg aagtacttgg
     2521 aggagagcaa ttttgtgcac agagatctgg ctgcaagaaa tgtgttgcta gttacccaac
     2581 attacgccaa gatcagtgat ttcggacttt ccaaagcact gcgtgctgat gaaaactact
     2641 acaaggccca gacccatgga aagtggcctg tcaagtggta cgctccggaa tgcatcaact
     2701 actacaagtt ctccagcaaa agcgatgtct ggagctttgg agtgttgatg tgggaagcat
     2761 tctcctatgg gcagaagcca tatcgaggga tgaaaggaag tgaagtcacc gctatgttag
     2821 agaaaggaga gcggatgggg tgccctgcag ggtgtccaag agagatgtac gatctcatga
     2881 atctgtgctg gacatacgat gtggaaaaca ggcccggatt cgcagcagtg gaactgcggc
     2941 tgcgcaatta ctactatgac gtggtgaacg tttctctcgg ctcgagcggc gtcttcacac
     3001 tcgaagattt cgttggggac tggcgacaga cagccggcta caacctggac caagtccttg
     3061 aacagggagg tgtgtccagt ttgtttcaga atctcggggt gtccgtaact ccgatccaaa
     3121 ggattgtcct gagcggtgaa aatgggctga agatcgacat ccatgtcatc atcccgtatg
     3181 aaggtctgag cggcgaccaa atgggccaga tcgaaaaaat ttttaaggtg gtgtaccctg
     3241 tggatgatca tcactttaag gtgatcctgc actatggcac actggtaatc gacggggtta
     3301 cgccgaacat gatcgactat ttcggacggc cgtatgaagg catcgccgtg ttcgacggca
     3361 aaaagatcac tgtaacaggg accctgtgga acggcaacaa aattatcgac gagcgcctga
     3421 tcaaccccga cggctccctg ctgttccgag taaccatcaa cggagtgacc ggctggcggc
     3481 tgtgcgaacg cattctggcg taaggccgcg actctagagt cgacctgcag gcatgcaagc
     3541 tgatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     3601 ataactagca taaccccttg gggcggccgc ttcgagcaga catgataaga tacattgatg
     3661 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     3721 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     3781 gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttta agcaagtaaa
     3841 acctctacaa atgtggtaaa atcgaatttt aacaaaatat taacgcttac aatttcctga
     3901 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgc ggatctgcgc
     3961 agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct tctgaggcgg
     4021 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     4081 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     4141 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     4201 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4261 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     4321 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttaatta
     4381 actgttgaca attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg
     4441 aactaaaccc aggaggcaga tcatgattga acaagatgga ttgcacgcag gttctccggc
     4501 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4561 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4621 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     4681 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4741 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4801 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4861 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4921 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4981 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     5041 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     5101 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     5161 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5221 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     5281 accgaccaag cgacgcccaa cctgccatca cgatggccgc aataaaatat ctttattttc
     5341 attacatctg tgtgttggtt ttttgtgtga atcgatagcg ataaggatcc tctttgcgct
     5401 tgcgttttcc cttgtccaga tagcccagta gctgacattc atccggggtc agcaccgttt
     5461 ctgcggactg gctttctacc cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5521 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5581 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5641 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5701 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5761 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5821 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5881 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5941 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6001 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     6061 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6121 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6181 aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6241 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6301 ccttttatag tccggaaata caggaacgca cgctggatgg cccttcgctg ggatggtgaa
     6361 accatgaaaa atggcagctt cagtggatta agtgggggta atgtggcctg taccctctgg
     6421 ttgcataggt attcatacgg ttaaaattta tcaggcgcga ttgcggcagt ttttcgggtg
     6481 gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc tgaacgcgtt ttagcggtgc
     6541 gtacaattaa gggattatgg taaatccact tactgtctgc cctcgtagcc atcgagataa
     6601 accgcagtac tccggccacg atgcgtccgg cgtagaggat cgagatct
//
