LOCUS       sequence-475312-        8095 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express NanoLuc-JAK1 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 8095)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8095)
  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            "Unpublished"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-JAK1 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..8095
                     /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(5174..5192)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    5229..5350
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5240..5259)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5294..5313
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5595..5952
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5595..5786
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5618..5638)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5803..5938
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5967..6014
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6046..6840
                     /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(6100..6119)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6710..6729
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6904..6952
                     /note="synthetic polyadenylation signal"
     rep_origin      7109..7654
                     /direction=RIGHT
                     /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     7555..7574
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7771..7788
                     /label=L4440
                     /note="L4440 vector, forward primer"
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 gcagtatcta aatataaaag
     1621 aggactgcaa tgccatggct ttctgtgcta aaatgaggag ctccaagaag actgaggtga
     1681 acctggaggc ccctgagcca ggggtggaag tgatcttcta tctgtcggac agggagcccc
     1741 tccggctggg cagtggagag tacacagcag aggaactgtg catcagggct gcacaggcat
     1801 gccgtatctc tcctctttgt cacaacctct ttgccctgta tgacgagaac accaagctct
     1861 ggtatgctcc aaatcgcacc atcaccgttg atgacaagat gtccctccgg ctccactacc
     1921 ggatgaggtt ctatttcacc aattggcatg gaaccaacga caatgagcag tcagtgtggc
     1981 gtcattctcc aaagaagcag aaaaatggct acgagaaaaa aaagattcca gatgcaaccc
     2041 ctctccttga tgccagctca ctggagtatc tgtttgctca gggacagtat gatttggtga
     2101 aatgcctggc tcctattcga gaccccaaga ccgagcagga tggacatgat attgagaacg
     2161 agtgtctagg gatggctgtc ctggccatct cacactatgc catgatgaag aagatgcagt
     2221 tgccagaact gcccaaggac atcagctaca agcgatatat tccagaaaca ttgaataagt
     2281 ccatcagaca gaggaacctt ctcaccagga tgcggataaa taatgttttc aaggatttcc
     2341 taaaggaatt taacaacaag accatttgtg acagcagcgt gtccacgcat gacctgaagg
     2401 tgaaatactt ggctaccttg gaaactttga caaaacatta cggtgctgaa atatttgaga
     2461 cttccatgtt actgatttca tcagaaaatg agatgaattg gtttcattcg aatgacggtg
     2521 gaaacgttct ctactacgaa gtgatggtga ctgggaatct tggaatccag tggaggcata
     2581 aaccaaatgt tgtttctgtt gaaaaggaaa aaaataaact gaagcggaaa aaactggaaa
     2641 ataaacacaa gaaggatgag gagaaaaaca agatccggga agagtggaac aatttttctt
     2701 acttccctga aatcactcac attgtaataa aggagtctgt ggtcagcatt aacaagcagg
     2761 acaacaagaa aatggaactg aagctctctt cccacgagga ggccttgtcc tttgtgtccc
     2821 tggtagatgg ctacttccgg ctcacagcag atgcccatca ttacctctgc accgacgtgg
     2881 cccccccgtt gatcgtccac aacatacaga atggctgtca tggtccaatc tgtacagaat
     2941 acgccatcaa taaattgcgg caagaaggaa gcgaggaggg gatgtacgtg ctgaggtgga
     3001 gctgcaccga ctttgacaac atcctcatga ccgtcacctg ctttgagaag tctgagcagg
     3061 tgcagggtgc ccagaagcag ttcaagaact ttcagatcga ggtgcagaag ggccgctaca
     3121 gtctgcacgg ttcggaccgc agcttcccca gcttgggaga cctcatgagc cacctcaaga
     3181 agcagatcct gcgcacggat aacatcagct tcatgctaaa acgctgctgc cagcccaagc
     3241 cccgagaaat ctccaacctg ctggtggcta ctaagaaagc ccaggagtgg cagcccgtct
     3301 accccatgag ccagctgagt ttcgatcgga tcctcaagaa ggatctggtg cagggcgagc
     3361 accttgggag aggcacgaga acacacatct attctgggac cctgatggat tacaaggatg
     3421 acgaaggaac ttctgaagag aagaagataa aagtgatcct caaagtctta gaccccagcc
     3481 acagggatat ttccctggcc ttcttcgagg cagccagcat gatgagacag gtctcccaca
     3541 aacacatcgt gtacctctat ggcgtctgtg tccgcgacgt ggagaatatc atggtggaag
     3601 agtttgtgga agggggtcct ctggatctct tcatgcaccg gaaaagcgat gtccttacca
     3661 caccatggaa attcaaagtt gccaaacagc tggccagtgc cctgagctac ttggaggata
     3721 aagacctggt ccatggaaat gtgtgtacta aaaacctcct cctggcccgt gagggcatcg
     3781 acagtgagtg tggcccattc atcaagctca gtgaccccgg catccccatt acggtgctgt
     3841 ctaggcaaga atgcattgaa cgaatcccat ggattgctcc tgagtgtgtt gaggactcca
     3901 agaacctgag tgtggctgct gacaagtgga gctttggaac cacgctctgg gaaatctgct
     3961 acaatggcga gatccccttg aaagacaaga cgctgattga gaaagagaga ttctatgaaa
     4021 gccggtgcag gccagtgaca ccatcatgta aggagctggc tgacctcatg acccgctgca
     4081 tgaactatga ccccaatcag aggcctttct tccgagccat catgagagac attaataagc
     4141 ttgaagagca gaatccagat attgtttcag aaaaaaaacc agcaactgaa gtggacccca
     4201 cacattttga aaagcgcttc ctaaagagga tccgtgactt gggagagggc cactttggga
     4261 aggttgagct ctgcaggtat gaccccgaag gggacaatac aggggagcag gtggctgtta
     4321 aatctctgaa gcctgagagt ggaggtaacc acatagctga tctgaaaaag gaaatcgaga
     4381 tcttaaggaa cctctatcat gagaacattg tgaagtacaa aggaatctgc acagaagacg
     4441 gaggaaatgg tattaagctc atcatggaat ttctgccttc gggaagcctt aaggaatatc
     4501 ttccaaagaa taagaacaaa ataaacctca aacagcagct aaaatatgcc gttcagattt
     4561 gtaaggggat ggactatttg ggttctcggc aatacgttca ccgggacttg gcagcaagaa
     4621 atgtccttgt tgagagtgaa caccaagtga aaattggaga cttcggttta accaaagcaa
     4681 ttgaaaccga taaggagtat tacaccgtca aggatgaccg ggacagccct gtgttttggt
     4741 atgctccaga atgtttaatg caatctaaat tttatattgc ctctgacgtc tggtcttttg
     4801 gagtcactct gcatgagctg ctgacttact gtgattcaga ttctagtccc atggctttgt
     4861 tcctgaaaat gataggccca acccatggcc agatgacagt cacaagactt gtgaatacgt
     4921 taaaagaagg aaaacgcctg ccgtgcccac ctaactgtcc agatgaggtt tatcaactta
     4981 tgaggaaatg ctgggaattc caaccatcca atcggacaag ctttcagaac cttattgaag
     5041 gatttgaagc acttttaaaa gtttaaacga attcgggctc ggtacccggg gatcctctag
     5101 agtcgacctg caggcatgca agctgatccg gctgctaaca aagcccgaaa ggaagctgag
     5161 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcggc cgcttcgagc
     5221 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     5281 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     5341 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg
     5401 ggaggttttt ttaagcaagt aaaacctcta caaatgtggt aaaatcgaat tttaacaaaa
     5461 tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     5521 acaccgcata cgcggatctg cgcagcacca tggcctgaaa taacctctga aagaggaact
     5581 tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     5641 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5701 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5761 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     5821 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5881 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     5941 ggcttttgca aaaagcttaa ttaactgttg acaattaatc atcggcatag tatatcggca
     6001 tagtataata cgacaaggtg aggaactaaa cccaggaggc agatcatgat tgaacaagat
     6061 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     6121 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     6181 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     6241 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     6301 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     6361 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     6421 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     6481 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     6541 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     6601 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     6661 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     6721 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     6781 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     6841 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgatggc
     6901 cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgata
     6961 gcgataagga tccgctcatg cgatgataag ctgtcaaaca tgagaattac aacttatatc
     7021 gtatggggct gacttcaggt gctacatttg aagagataaa ttgcactgaa atctagaaat
     7081 attttatctg attaataaga tgatcttctt gagatcgttt tggtctgcgc gtaatctctt
     7141 gctctgaaaa cgaaaaaacc gccttgcagg gcggtttttc gaaggttctc tgagctacca
     7201 actctttgaa ccgaggtaac tggcttggag gagcgcagtc accaaaactt gtcctttcag
     7261 tttagcctta accggcgcat gacttcaaga ctaactcctc taaatcaatt accagtggct
     7321 gctgccagtg gtgcttttgc atgtctttcc gggttggact caagacgata gttaccggat
     7381 aaggcgcagc ggtcggactg aacggggggt tcgtgcatac agtccagctt ggagcgaact
     7441 gcctacccgg aactgagtgt caggcgtgga atgagacaaa cgcggccata acagcggaat
     7501 gacaccggta aaccgaaagg caggaacagg agagcgcacg agggagccgc cagggggaaa
     7561 cgcctggtat ctttatagtc ctgtcgggtt tcgccaccac tgatttgagc gtcagatttc
     7621 gtgatgcttg tcaggggggc ggagcctatg gaaaaacggc tttgccgcgg ccctctcact
     7681 tccctgttaa gtatcttcct ggcatcttcc aggaaatctc cgccccgttc gtaagccatt
     7741 tccgctcgcc gcagtcgaac gaccgagcgt agcgagtcag tgagcgagga agcggaatat
     7801 atcctgtatc acatattctg ctgacgcacc ggtgcagcct tttttctcct gccacatgaa
     7861 gcacttcact gacaccctca tcagtgccaa catagtaagc cagtatacac tccgctagcg
     7921 ctgatgtccg gcggtgcttt tgccgttacg caccaccccg tcatgagatt acgcgtttta
     7981 gcggtgcgta caattaaggg attatggtaa atccacttac tgtctgccct cgtagccatc
     8041 gagataaacc gcagtactcc ggccacgatg cgtccggcgt agaggatcga gatct
//
