LOCUS       sequence-486457-        8069 bp    DNA     circular SYN 17-SEP-2025
DEFINITION  Express JAK1-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 8069)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8069)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: JAK1-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..8069
                     /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             4548..5060
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(5148..5166)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    5203..5324
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5214..5233)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5268..5287
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5569..5926
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5569..5760
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5592..5612)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5777..5912
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5941..5988
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6020..6814
                     /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(6074..6093)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6684..6703
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6878..6926
                     /note="synthetic polyadenylation signal"
     rep_origin      7083..7628
                     /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     7529..7548
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7745..7762
                     /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 ggctagcgat cgccatgcag tatctaaata
     1081 taaaagagga ctgcaatgcc atggctttct gtgctaaaat gaggagctcc aagaagactg
     1141 aggtgaacct ggaggcccct gagccagggg tggaagtgat cttctatctg tcggacaggg
     1201 agcccctccg gctgggcagt ggagagtaca cagcagagga actgtgcatc agggctgcac
     1261 aggcatgccg tatctctcct ctttgtcaca acctctttgc cctgtatgac gagaacacca
     1321 agctctggta tgctccaaat cgcaccatca ccgttgatga caagatgtcc ctccggctcc
     1381 actaccggat gaggttctat ttcaccaatt ggcatggaac caacgacaat gagcagtcag
     1441 tgtggcgtca ttctccaaag aagcagaaaa atggctacga gaaaaaaaag attccagatg
     1501 caacccctct ccttgatgcc agctcactgg agtatctgtt tgctcaggga cagtatgatt
     1561 tggtgaaatg cctggctcct attcgagacc ccaagaccga gcaggatgga catgatattg
     1621 agaacgagtg tctagggatg gctgtcctgg ccatctcaca ctatgccatg atgaagaaga
     1681 tgcagttgcc agaactgccc aaggacatca gctacaagcg atatattcca gaaacattga
     1741 ataagtccat cagacagagg aaccttctca ccaggatgcg gataaataat gttttcaagg
     1801 atttcctaaa ggaatttaac aacaagacca tttgtgacag cagcgtgtcc acgcatgacc
     1861 tgaaggtgaa atacttggct accttggaaa ctttgacaaa acattacggt gctgaaatat
     1921 ttgagacttc catgttactg atttcatcag aaaatgagat gaattggttt cattcgaatg
     1981 acggtggaaa cgttctctac tacgaagtga tggtgactgg gaatcttgga atccagtgga
     2041 ggcataaacc aaatgttgtt tctgttgaaa aggaaaaaaa taaactgaag cggaaaaaac
     2101 tggaaaataa acacaagaag gatgaggaga aaaacaagat ccgggaagag tggaacaatt
     2161 tttcttactt ccctgaaatc actcacattg taataaagga gtctgtggtc agcattaaca
     2221 agcaggacaa caagaaaatg gaactgaagc tctcttccca cgaggaggcc ttgtcctttg
     2281 tgtccctggt agatggctac ttccggctca cagcagatgc ccatcattac ctctgcaccg
     2341 acgtggcccc cccgttgatc gtccacaaca tacagaatgg ctgtcatggt ccaatctgta
     2401 cagaatacgc catcaataaa ttgcggcaag aaggaagcga ggaggggatg tacgtgctga
     2461 ggtggagctg caccgacttt gacaacatcc tcatgaccgt cacctgcttt gagaagtctg
     2521 agcaggtgca gggtgcccag aagcagttca agaactttca gatcgaggtg cagaagggcc
     2581 gctacagtct gcacggttcg gaccgcagct tccccagctt gggagacctc atgagccacc
     2641 tcaagaagca gatcctgcgc acggataaca tcagcttcat gctaaaacgc tgctgccagc
     2701 ccaagccccg agaaatctcc aacctgctgg tggctactaa gaaagcccag gagtggcagc
     2761 ccgtctaccc catgagccag ctgagtttcg atcggatcct caagaaggat ctggtgcagg
     2821 gcgagcacct tgggagaggc acgagaacac acatctattc tgggaccctg atggattaca
     2881 aggatgacga aggaacttct gaagagaaga agataaaagt gatcctcaaa gtcttagacc
     2941 ccagccacag ggatatttcc ctggccttct tcgaggcagc cagcatgatg agacaggtct
     3001 cccacaaaca catcgtgtac ctctatggcg tctgtgtccg cgacgtggag aatatcatgg
     3061 tggaagagtt tgtggaaggg ggtcctctgg atctcttcat gcaccggaaa agcgatgtcc
     3121 ttaccacacc atggaaattc aaagttgcca aacagctggc cagtgccctg agctacttgg
     3181 aggataaaga cctggtccat ggaaatgtgt gtactaaaaa cctcctcctg gcccgtgagg
     3241 gcatcgacag tgagtgtggc ccattcatca agctcagtga ccccggcatc cccattacgg
     3301 tgctgtctag gcaagaatgc attgaacgaa tcccatggat tgctcctgag tgtgttgagg
     3361 actccaagaa cctgagtgtg gctgctgaca agtggagctt tggaaccacg ctctgggaaa
     3421 tctgctacaa tggcgagatc cccttgaaag acaagacgct gattgagaaa gagagattct
     3481 atgaaagccg gtgcaggcca gtgacaccat catgtaagga gctggctgac ctcatgaccc
     3541 gctgcatgaa ctatgacccc aatcagaggc ctttcttccg agccatcatg agagacatta
     3601 ataagcttga agagcagaat ccagatattg tttcagaaaa aaaaccagca actgaagtgg
     3661 accccacaca ttttgaaaag cgcttcctaa agaggatccg tgacttggga gagggccact
     3721 ttgggaaggt tgagctctgc aggtatgacc ccgaagggga caatacaggg gagcaggtgg
     3781 ctgttaaatc tctgaagcct gagagtggag gtaaccacat agctgatctg aaaaaggaaa
     3841 tcgagatctt aaggaacctc tatcatgaga acattgtgaa gtacaaagga atctgcacag
     3901 aagacggagg aaatggtatt aagctcatca tggaatttct gccttcggga agccttaagg
     3961 aatatcttcc aaagaataag aacaaaataa acctcaaaca gcagctaaaa tatgccgttc
     4021 agatttgtaa ggggatggac tatttgggtt ctcggcaata cgttcaccgg gacttggcag
     4081 caagaaatgt ccttgttgag agtgaacacc aagtgaaaat tggagacttc ggtttaacca
     4141 aagcaattga aaccgataag gagtattaca ccgtcaagga tgaccgggac agccctgtgt
     4201 tttggtatgc tccagaatgt ttaatgcaat ctaaatttta tattgcctct gacgtctggt
     4261 cttttggagt cactctgcat gagctgctga cttactgtga ttcagattct agtcccatgg
     4321 ctttgttcct gaaaatgata ggcccaaccc atggccagat gacagtcaca agacttgtga
     4381 atacgttaaa agaaggaaaa cgcctgccgt gcccacctaa ctgtccagat gaggtttatc
     4441 aacttatgag gaaatgctgg gaattccaac catccaatcg gacaagcttt cagaacctta
     4501 ttgaaggatt tgaagcactt ttaaaagttt ctctcggctc gagcggcgtc ttcacactcg
     4561 aagatttcgt tggggactgg cgacagacag ccggctacaa cctggaccaa gtccttgaac
     4621 agggaggtgt gtccagtttg tttcagaatc tcggggtgtc cgtaactccg atccaaagga
     4681 ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca tgtcatcatc ccgtatgaag
     4741 gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt taaggtggtg taccctgtgg
     4801 atgatcatca ctttaaggtg atcctgcact atggcacact ggtaatcgac ggggttacgc
     4861 cgaacatgat cgactatttc ggacggccgt atgaaggcat cgccgtgttc gacggcaaaa
     4921 agatcactgt aacagggacc ctgtggaacg gcaacaaaat tatcgacgag cgcctgatca
     4981 accccgacgg ctccctgctg ttccgagtaa ccatcaacgg agtgaccggc tggcggctgt
     5041 gcgaacgcat tctggcgtaa ggccgcgact ctagagtcga cctgcaggca tgcaagctga
     5101 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
     5161 actagcataa ccccttgggg cggccgcttc gagcagacat gataagatac attgatgagt
     5221 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     5281 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     5341 ttcattttat gtttcaggtt cagggggaga tgtgggaggt ttttttaagc aagtaaaacc
     5401 tctacaaatg tggtaaaatc gaattttaac aaaatattaa cgcttacaat ttcctgatgc
     5461 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgcgga tctgcgcagc
     5521 accatggcct gaaataacct ctgaaagagg aacttggtta ggtaccttct gaggcggaaa
     5581 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     5641 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     5701 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     5761 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     5821 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     5881 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttaattaact
     5941 gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac
     6001 taaacccagg aggcagatca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     6061 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     6121 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     6181 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     6241 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     6301 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     6361 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     6421 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     6481 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     6541 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     6601 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     6661 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     6721 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     6781 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     6841 gaccaagcga cgcccaacct gccatcacga tggccgcaat aaaatatctt tattttcatt
     6901 acatctgtgt gttggttttt tgtgtgaatc gatagcgata aggatccgct catgcgatga
     6961 taagctgtca aacatgagaa ttacaactta tatcgtatgg ggctgacttc aggtgctaca
     7021 tttgaagaga taaattgcac tgaaatctag aaatatttta tctgattaat aagatgatct
     7081 tcttgagatc gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg
     7141 cagggcggtt tttcgaaggt tctctgagct accaactctt tgaaccgagg taactggctt
     7201 ggaggagcgc agtcaccaaa acttgtcctt tcagtttagc cttaaccggc gcatgacttc
     7261 aagactaact cctctaaatc aattaccagt ggctgctgcc agtggtgctt ttgcatgtct
     7321 ttccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg actgaacggg
     7381 gggttcgtgc atacagtcca gcttggagcg aactgcctac ccggaactga gtgtcaggcg
     7441 tggaatgaga caaacgcggc cataacagcg gaatgacacc ggtaaaccga aaggcaggaa
     7501 caggagagcg cacgagggag ccgccagggg gaaacgcctg gtatctttat agtcctgtcg
     7561 ggtttcgcca ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc
     7621 tatggaaaaa cggctttgcc gcggccctct cacttccctg ttaagtatct tcctggcatc
     7681 ttccaggaaa tctccgcccc gttcgtaagc catttccgct cgccgcagtc gaacgaccga
     7741 gcgtagcgag tcagtgagcg aggaagcgga atatatcctg tatcacatat tctgctgacg
     7801 caccggtgca gccttttttc tcctgccaca tgaagcactt cactgacacc ctcatcagtg
     7861 ccaacatagt aagccagtat acactccgct agcgctgatg tccggcggtg cttttgccgt
     7921 tacgcaccac cccgtcatga gattacgcgt tttagcggtg cgtacaatta agggattatg
     7981 gtaaatccac ttactgtctg ccctcgtagc catcgagata aaccgcagta ctccggccac
     8041 gatgcgtccg gcgtagagga tcgagatct
//
