LOCUS       sequence-479805-        8330 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express NanoLuc-TYK2 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 8330)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8330)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoLuc-TYK2 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..8330
                     /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(5273..5291)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    5328..5449
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5339..5358)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5393..5412
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5694..6051
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5694..5885
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5717..5737)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5902..6037
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6066..6113
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6145..6939
                     /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(6199..6218)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6809..6828
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7003..7051
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(7290..7878)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7370..7389)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7994..8277
                     /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 gcctctgcgc cactggggga
     1621 tggccagggg cagtaagccc gttggggatg gagcccagcc catggctgcc atgggaggcc
     1681 tgaaggtgct tctgcactgg gctggtccag gcggcgggga gccctgggtc actttcagtg
     1741 agtcatcgct gacagctgag gaagtctgca tccacattgc acataaagtt ggtatcactc
     1801 ctccttgctt caatctcttt gccctcttcg atgctcaggc ccaagtctgg ttgcccccaa
     1861 accacatcct agagatcccc agagatgcaa gcctgatgct atatttccgc ataaggtttt
     1921 atttccggaa ctggcatggc atgaatcctc gggaaccggc tgtgtaccgt tgtgggcccc
     1981 caggaaccga ggcatcctca gatcagacag cacaggggat gcaactcctg gacccagcct
     2041 catttgagta cctctttgag cagggcaagc atgagtttgt gaatgacgtg gcatcactgt
     2101 gggagctgtc gaccgaggag gagatccacc actttaagaa tgagagcctg ggcatggcct
     2161 ttctgcacct ctgtcacctc gctctccgcc atggcatccc cctggaggag gtggccaaga
     2221 agaccagctt caaggactgc atcccgcgct ccttccgccg gcatatccgg cagcacagcg
     2281 ccctgacccg gctgcgcctt cggaacgtct tccgcaggtt cctgcgggac ttccagccgg
     2341 gccgactctc ccagcagatg gtcatggtca aatacctagc cacactcgag cggctggcac
     2401 cccgcttcgg cacagagcgt gtgcccgtgt gccacctgag gctgctggcc caggccgagg
     2461 gggagccctg ctacatccgg gacagtgggg tggcccctac agaccctggc cctgagtctg
     2521 ctgctgggcc cccaacccac gaggtgctgg tgacaggcac tggtggcatc cagtggtggc
     2581 cagtagagga ggaggtgaac aaggaggagg gttctagtgg cagcagtggc aggaaccccc
     2641 aagccagcct gtttgggaag aaggccaagg ctcacaaggc agtcggccag ccggcagaca
     2701 ggccgcggga gccactgtgg gcctacttct gtgacttccg ggacatcacc cacgtggtgc
     2761 tgaaagagca ctgtgtcagc atccaccggc aggacaacaa gtgcctggag ctgagcttgc
     2821 cttcccgggc tgcggcgctg tccttcgtgt cgctggtgga cggctatttc cgcctgacgg
     2881 ccgactccag ccactacctg tgccacgagg tggctccccc acggctggtg atgagcatcc
     2941 gggatgggat ccacggaccc ctgctggagc catttgtgca ggccaagctg cggcccgagg
     3001 acggcctgta cctcattcac tggagcacca gccaccccta ccgcctgatc ctcacagtgg
     3061 cccagcgtag ccaggcacca gacggcatgc agagcttgcg gctccgaaag ttccccattg
     3121 agcagcagga cggggccttc gtgctggagg gctggggccg gtccttcccc agcgttcggg
     3181 aacttggggc tgccttgcag ggctgcttgc tgagggccgg ggatgactgc ttctctctgc
     3241 gtcgctgttg cctgccccaa ccaggagaaa cctccaatct catcatcatg cggggggctc
     3301 gggccagccc caggacactc aacctcagcc agctcagctt ccaccgggtt gaccagaagg
     3361 agatcaccca gctgtcccac ttgggccagg gcacaaggac caacgtgtat gagggccgcc
     3421 tgcgagtgga gggcagcggg gaccctgagg agggcaagat ggatgacgag gaccccctcg
     3481 tgcctggcag ggaccgtggg caggagctac gagtggtgct caaagtgctg gaccctagtc
     3541 accatgacat cgccctggcc ttctacgaga cagccagcct catgagccag gtctcccaca
     3601 cgcacctggc cttcgtgcat ggcgtctgtg tgcgcggccc tgaaaatatc atggtgacag
     3661 agtacgtgga gcacggaccc ctggatgtgt ggctgcggag ggagcggggc catgtgccca
     3721 tggcttggaa gatggtggtg gcccagcagc tggccagcgc cctcagctac ctggagaaca
     3781 agaacctggt tcatggtaat gtgtgtggcc ggaacatcct gctggcccgg ctggggttgg
     3841 cagagggcac cagccccttc atcaagctga gtgatcctgg cgtgggcctg ggcgccctct
     3901 ccagggagga gcgggtggag aggatcccct ggctggcccc cgaatgccta ccaggtgggg
     3961 ccaacagcct aagcaccgcc atggacaagt gggggtttgg cgccaccctc ctggagatct
     4021 gctttgacgg agaggcccct ctgcagagcc gcagtccctc cgagaaggag catttctacc
     4081 agaggcagca ccggctgccc gagccctcct gcccacagct ggccacactc accagccagt
     4141 gtctgaccta tgagccaacc cagaggccat cattccgcac catcctgcgt gacctcaccc
     4201 ggctgcagcc ccacaatctt gctgacgtct tgactgtgaa cccggactca ccggcgtcgg
     4261 accctacggt tttccacaag cgctatttga aaaagatccg agatctgggc gagggtcact
     4321 tcggcaaggt cagcttgtac tgctacgatc cgaccaacga cggcactggc gagatggtgg
     4381 cggtgaaagc cctcaaggca gactgcggcc cccagcaccg ctcgggctgg aagcaggaga
     4441 ttgacattct gcgcacgctc taccacgagc acatcatcaa gtacaagggc tgctgcgagg
     4501 accaaggcga gaagtcgctg cagctggtca tggagtacgt gcccctgggc agcctccgag
     4561 actacctgcc ccggcacagc atcgggctgg cccagctgct gctcttcgcc cagcagatct
     4621 gcgagggcat ggcctatctg cacgcgcagc actacatcca ccgagaccta gccgcgcgca
     4681 acgtgctgct ggacaacgac aggctggtca agatcgggga ctttggccta gccaaggccg
     4741 tgcccgaagg ccacgagtac taccgcgtgc gcgaggatgg ggacagcccc gtgttctggt
     4801 atgccccaga gtgcctgaag gagtataagt tctactatgc gtcagatgtc tggtccttcg
     4861 gggtgaccct gtatgagctg ctgacgcact gtgactccag ccagagcccc cccacgaaat
     4921 tccttgagct cataggcatt gctcagggtc agatgacagt tctgagactc actgagttgc
     4981 tggaacgagg ggagaggctg ccacggcccg acaaatgtcc ctgtgaggtc tatcatctca
     5041 tgaagaactg ctgggagaca gaggcgtcct ttcgcccaac cttcgagaac ctcataccca
     5101 ttctgaagac agtccatgag aagtaccaag gccaggcccc ttcagtgttc agcgtgtgcg
     5161 tttaaacgaa ttcgggctcg gtacccgggg atcctctaga gtcgacctgc aggcatgcaa
     5221 gctgatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
     5281 caataactag cataacccct tggggcggcc gcttcgagca gacatgataa gatacattga
     5341 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     5401 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     5461 ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt taagcaagta
     5521 aaacctctac aaatgtggta aaatcgaatt ttaacaaaat attaacgctt acaatttcct
     5581 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gcggatctgc
     5641 gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttaggtac cttctgaggc
     5701 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     5761 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     5821 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     5881 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     5941 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     6001 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagcttaat
     6061 taactgttga caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga
     6121 ggaactaaac ccaggaggca gatcatgatt gaacaagatg gattgcacgc aggttctccg
     6181 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     6241 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     6301 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     6361 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     6421 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     6481 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     6541 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     6601 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     6661 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     6721 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     6781 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     6841 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     6901 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     6961 tgaccgacca agcgacgccc aacctgccat cacgatggcc gcaataaaat atctttattt
     7021 tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag cgataaggat cctctttgcg
     7081 cttgcgtttt cccttgtcca gatagcccag tagctgacat tcatccgggg tcagcaccgt
     7141 ttctgcggac tggctttcta cccggtatca gctcactcaa aggcggtaat acggttatcc
     7201 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     7261 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     7321 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     7381 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     7441 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     7501 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     7561 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     7621 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     7681 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     7741 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     7801 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     7861 agaaaaaaag gatttcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     7921 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     7981 atccttttat agtccggaaa tacaggaacg cacgctggat ggcccttcgc tgggatggtg
     8041 aaaccatgaa aaatggcagc ttcagtggat taagtggggg taatgtggcc tgtaccctct
     8101 ggttgcatag gtattcatac ggttaaaatt tatcaggcgc gattgcggca gtttttcggg
     8161 tggtttgttg ccatttttac ctgtctgctg ccgtgatcgc gctgaacgcg ttttagcggt
     8221 gcgtacaatt aagggattat ggtaaatcca cttactgtct gccctcgtag ccatcgagat
     8281 aaaccgcagt actccggcca cgatgcgtcc ggcgtagagg atcgagatct
//
