LOCUS       sequence-479785-        7979 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express JAK3-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 7979)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7979)
  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            "Unpublished"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: JAK3-NanoLuc Vector
FEATURES             Location/Qualifiers
     source          1..7979
                     /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             4458..4970
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(5058..5076)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    5113..5234
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5124..5143)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5178..5197
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5479..5836
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5479..5670
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5502..5522)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5687..5822
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5851..5898
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5930..6724
                     /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(5984..6003)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6594..6613
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6788..6836
                     /note="synthetic polyadenylation signal"
     rep_origin      6993..7538
                     /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     7439..7458
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7655..7672
                     /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 cgccatggca cctccaagtg
     1081 aagagacgcc cctgatccct cagcgttcat gcagcctctt gtccacggag gctggtgccc
     1141 tgcatgtgct gctgcccgct cggggccccg ggccccccca gcgcctatct ttctcctttg
     1201 gggaccactt ggctgaggac ctgtgcgtgc aggctgccaa ggccagcggc atcctgcctg
     1261 tgtaccactc cctctttgct ctggccacgg aggacctgtc ctgctggttc cccccgagcc
     1321 acatcttctc cgtggaggat gccagcaccc aagtcctgct gtacaggatt cgcttttact
     1381 tccccaattg gtttgggctg gagaagtgcc accgcttcgg gctacgcaag gatttggcca
     1441 gtgctatcct tgacctgcca gtcctggagc acctctttgc ccagcaccgc agtgacctgg
     1501 tgagtgggcg cctccccgtg ggcctcagtc tcaaggagca gggtgagtgt ctcagcctgg
     1561 ccgtgttgga cctggcccgg atggcgcgag agcaggccca gcggccggga gagctgctga
     1621 agactgtcag ctacaaggcc tgcctacccc caagcctgcg cgacctgatc cagggcctga
     1681 gcttcgtgac gcggaggcgt attcggagga cggtgcgcag agccctgcgc cgcgtggccg
     1741 cctgccaggc agaccggcac tcgctcatgg ccaagtacat catggacctg gagcggctgg
     1801 atccagccgg ggccgccgag accttccacg tgggcctccc tggggccctt ggtggccacg
     1861 acgggctggg gctgctccgc gtggctggtg acggcggcat cgcctggacc cagggagaac
     1921 aggaggtcct ccagcccttc tgcgactttc cagaaatcgt agacattagc atcaagcagg
     1981 ccccgcgcgt tggcccggcc ggagagcacc gcctggtcac tgttaccagg acagacaacc
     2041 agattttaga ggccgagttc ccagggctgc ccgaggctct gtcgttcgtg gcgctcgtgg
     2101 acggctactt ccggctgacc acggactccc agcacttctt ctgcaaggag gtggcaccgc
     2161 cgaggctgct ggaggaagtg gccgagcagt gccacggccc catcactctg gactttgcca
     2221 tcaacaagct caagactggg ggctcacgtc ctggctccta tgttctccgc cgcagccccc
     2281 aggactttga cagcttcctc ctcactgtct gtgtccagaa cccccttggt cctgattata
     2341 agggctgcct catccggcgc agccccacag gaaccttcct tctggttggc ctcagccgac
     2401 cccacagcag tcttcgagag ctcctggcaa cctgctggga tggggggctg cacgtagatg
     2461 gggtggcagt gaccctcact tcctgctgta tccccagacc caaagaaaag tccaacctga
     2521 tcgtggtcca gagaggtcac agcccaccca catcatcctt ggttcagccc caatcccaat
     2581 accagctgag tcagatgaca tttcacaaga tccctgctga cagcctggag tggcatgaga
     2641 acctgggcca tgggtccttc accaagattt accggggctg tcgccatgag gtggtggatg
     2701 gggaggcccg aaagacagag gtgctgctga aggtcatgga tgccaagcac aagaactgca
     2761 tggagtcatt cctggaagca gcgagcttga tgagccaagt gtcgtaccgg catctcgtgc
     2821 tgctccacgg cgtgtgcatg gctggagaca gcaccatggt gcaggaattt gtacacctgg
     2881 gggccataga catgtatctg cgaaaacgtg gccacctggt gccagccagc tggaagctgc
     2941 aggtggtcaa acagctggcc tacgccctca actatctgga ggacaaaggc ctgccccatg
     3001 gcaatgtctc tgcccggaag gtgctcctgg ctcgggaggg ggctgatggg agcccgccct
     3061 tcatcaagct gagtgaccct ggggtcagcc ccgctgtgtt aagcctggag atgctcaccg
     3121 acaggatccc ctgggtggcc cccgagtgtc tccgggaggc gcagacactt agcttggaag
     3181 ctgacaagtg gggcttcggc gccacggtct gggaagtgtt tagtggcgtc accatgccca
     3241 tcagtgccct ggatcctgct aagaaactcc aattttatga ggaccggcag cagctgccgg
     3301 cccccaagtg gacagagctg gccctgctga ttcaacagtg catggcctat gagccggtcc
     3361 agaggccctc cttccgagcc gtcattcgtg acctcaatag cctcatctct tcagactatg
     3421 agctcctctc agaccccaca cctggtgccc tggcacctcg tgatgggctg tggaatggtg
     3481 cccagctcta tgcctgccaa gaccccacga tcttcgagga gagacacctc aagtacatct
     3541 cacagctggg caagggcaac tttggcagcg tggagctgtg ccgctatgac ccgctaggcg
     3601 acaatacagg tgccctggtg gccgtgaaac agctgcagca cagcgggcca gaccagcaga
     3661 gggactttca gcgggagatt cagatcctca aagcactgca cagtgatttc attgtcaagt
     3721 atcgtggtgt cagctatggc ccgggccgcc agagcctgcg gctggtcatg gagtacctgc
     3781 ccagcggctg cttgcgcgac ttcctgcagc ggcaccgcgc gcgcctcgat gccagccgcc
     3841 tccttctcta ttcctcgcag atctgcaagg gcatggagta cctgggctcc cgccgctgcg
     3901 tgcaccgcga cctggccgcc cgaaacatcc tcgtggagag cgaggcacac gtcaagatcg
     3961 ctgacttcgg cctagctaag ctgctgccgc ttgacaaaga ctactacgtg gtccgcgagc
     4021 caggccagag ccccattttc tggtatgccc ccgaatccct ctcggacaac atcttctctc
     4081 gccagtcaga cgtctggagc ttcggggtcg tcctgtacga gctcttcacc tactgcgaca
     4141 aaagctgcag cccctcggcc gagttcctgc ggatgatggg atgtgagcgg gatgtccccg
     4201 ccctctgccg cctcttggaa ctgctggagg agggccagag gctgccggcg cctcctgcct
     4261 gccctgctga ggttcacgag ctcatgaagc tgtgctgggc ccctagccca caggaccggc
     4321 catcattcag cgccctgggc ccccagctgg acatgctgtg gagcggaagc cgggggtgtg
     4381 agactcatgc cttcactgct cacccagagg gcaaacacca ctccctgtcc ttttcagttt
     4441 ctctcggctc gagcggcgtc ttcacactcg aagatttcgt tggggactgg cgacagacag
     4501 ccggctacaa cctggaccaa gtccttgaac agggaggtgt gtccagtttg tttcagaatc
     4561 tcggggtgtc cgtaactccg atccaaagga ttgtcctgag cggtgaaaat gggctgaaga
     4621 tcgacatcca tgtcatcatc ccgtatgaag gtctgagcgg cgaccaaatg ggccagatcg
     4681 aaaaaatttt taaggtggtg taccctgtgg atgatcatca ctttaaggtg atcctgcact
     4741 atggcacact ggtaatcgac ggggttacgc cgaacatgat cgactatttc ggacggccgt
     4801 atgaaggcat cgccgtgttc gacggcaaaa agatcactgt aacagggacc ctgtggaacg
     4861 gcaacaaaat tatcgacgag cgcctgatca accccgacgg ctccctgctg ttccgagtaa
     4921 ccatcaacgg agtgaccggc tggcggctgt gcgaacgcat tctggcgtaa ggccgcgact
     4981 ctagagtcga cctgcaggca tgcaagctga tccggctgct aacaaagccc gaaaggaagc
     5041 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cggccgcttc
     5101 gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     5161 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     5221 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga
     5281 tgtgggaggt ttttttaagc aagtaaaacc tctacaaatg tggtaaaatc gaattttaac
     5341 aaaatattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta
     5401 tttcacaccg catacgcgga tctgcgcagc accatggcct gaaataacct ctgaaagagg
     5461 aacttggtta ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     5521 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     5581 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     5641 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     5701 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     5761 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     5821 cctaggcttt tgcaaaaagc ttaattaact gttgacaatt aatcatcggc atagtatatc
     5881 ggcatagtat aatacgacaa ggtgaggaac taaacccagg aggcagatca tgattgaaca
     5941 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     6001 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     6061 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     6121 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     6181 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     6241 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     6301 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     6361 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     6421 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     6481 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     6541 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     6601 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     6661 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     6721 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     6781 tggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgaatc
     6841 gatagcgata aggatccgct catgcgatga taagctgtca aacatgagaa ttacaactta
     6901 tatcgtatgg ggctgacttc aggtgctaca tttgaagaga taaattgcac tgaaatctag
     6961 aaatatttta tctgattaat aagatgatct tcttgagatc gttttggtct gcgcgtaatc
     7021 tcttgctctg aaaacgaaaa aaccgccttg cagggcggtt tttcgaaggt tctctgagct
     7081 accaactctt tgaaccgagg taactggctt ggaggagcgc agtcaccaaa acttgtcctt
     7141 tcagtttagc cttaaccggc gcatgacttc aagactaact cctctaaatc aattaccagt
     7201 ggctgctgcc agtggtgctt ttgcatgtct ttccgggttg gactcaagac gatagttacc
     7261 ggataaggcg cagcggtcgg actgaacggg gggttcgtgc atacagtcca gcttggagcg
     7321 aactgcctac ccggaactga gtgtcaggcg tggaatgaga caaacgcggc cataacagcg
     7381 gaatgacacc ggtaaaccga aaggcaggaa caggagagcg cacgagggag ccgccagggg
     7441 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca ccactgattt gagcgtcaga
     7501 tttcgtgatg cttgtcaggg gggcggagcc tatggaaaaa cggctttgcc gcggccctct
     7561 cacttccctg ttaagtatct tcctggcatc ttccaggaaa tctccgcccc gttcgtaagc
     7621 catttccgct cgccgcagtc gaacgaccga gcgtagcgag tcagtgagcg aggaagcgga
     7681 atatatcctg tatcacatat tctgctgacg caccggtgca gccttttttc tcctgccaca
     7741 tgaagcactt cactgacacc ctcatcagtg ccaacatagt aagccagtat acactccgct
     7801 agcgctgatg tccggcggtg cttttgccgt tacgcaccac cccgtcatga gattacgcgt
     7861 tttagcggtg cgtacaatta agggattatg gtaaatccac ttactgtctg ccctcgtagc
     7921 catcgagata aaccgcagta ctccggccac gatgcgtccg gcgtagagga tcgagatct
//
