LOCUS       sequence-479786-        8005 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express NanoLuc-JAK3 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 8005)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8005)
  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            "Unpublished"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-JAK3 Vector
FEATURES             Location/Qualifiers
     source          1..8005
                     /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(5084..5102)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    5139..5260
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5150..5169)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5204..5223
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5505..5862
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5505..5696
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5528..5548)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5713..5848
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5877..5924
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5956..6750
                     /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(6010..6029)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6620..6639
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6814..6862
                     /note="synthetic polyadenylation signal"
     rep_origin      7019..7564
                     /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     7465..7484
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7681..7698
                     /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 ggcacctcca agtgaagaga
     1621 cgcccctgat ccctcagcgt tcatgcagcc tcttgtccac ggaggctggt gccctgcatg
     1681 tgctgctgcc cgctcggggc cccgggcccc cccagcgcct atctttctcc tttggggacc
     1741 acttggctga ggacctgtgc gtgcaggctg ccaaggccag cggcatcctg cctgtgtacc
     1801 actccctctt tgctctggcc acggaggacc tgtcctgctg gttccccccg agccacatct
     1861 tctccgtgga ggatgccagc acccaagtcc tgctgtacag gattcgcttt tacttcccca
     1921 attggtttgg gctggagaag tgccaccgct tcgggctacg caaggatttg gccagtgcta
     1981 tccttgacct gccagtcctg gagcacctct ttgcccagca ccgcagtgac ctggtgagtg
     2041 ggcgcctccc cgtgggcctc agtctcaagg agcagggtga gtgtctcagc ctggccgtgt
     2101 tggacctggc ccggatggcg cgagagcagg cccagcggcc gggagagctg ctgaagactg
     2161 tcagctacaa ggcctgccta cccccaagcc tgcgcgacct gatccagggc ctgagcttcg
     2221 tgacgcggag gcgtattcgg aggacggtgc gcagagccct gcgccgcgtg gccgcctgcc
     2281 aggcagaccg gcactcgctc atggccaagt acatcatgga cctggagcgg ctggatccag
     2341 ccggggccgc cgagaccttc cacgtgggcc tccctggggc ccttggtggc cacgacgggc
     2401 tggggctgct ccgcgtggct ggtgacggcg gcatcgcctg gacccaggga gaacaggagg
     2461 tcctccagcc cttctgcgac tttccagaaa tcgtagacat tagcatcaag caggccccgc
     2521 gcgttggccc ggccggagag caccgcctgg tcactgttac caggacagac aaccagattt
     2581 tagaggccga gttcccaggg ctgcccgagg ctctgtcgtt cgtggcgctc gtggacggct
     2641 acttccggct gaccacggac tcccagcact tcttctgcaa ggaggtggca ccgccgaggc
     2701 tgctggagga agtggccgag cagtgccacg gccccatcac tctggacttt gccatcaaca
     2761 agctcaagac tgggggctca cgtcctggct cctatgttct ccgccgcagc ccccaggact
     2821 ttgacagctt cctcctcact gtctgtgtcc agaaccccct tggtcctgat tataagggct
     2881 gcctcatccg gcgcagcccc acaggaacct tccttctggt tggcctcagc cgaccccaca
     2941 gcagtcttcg agagctcctg gcaacctgct gggatggggg gctgcacgta gatggggtgg
     3001 cagtgaccct cacttcctgc tgtatcccca gacccaaaga aaagtccaac ctgatcgtgg
     3061 tccagagagg tcacagccca cccacatcat ccttggttca gccccaatcc caataccagc
     3121 tgagtcagat gacatttcac aagatccctg ctgacagcct ggagtggcat gagaacctgg
     3181 gccatgggtc cttcaccaag atttaccggg gctgtcgcca tgaggtggtg gatggggagg
     3241 cccgaaagac agaggtgctg ctgaaggtca tggatgccaa gcacaagaac tgcatggagt
     3301 cattcctgga agcagcgagc ttgatgagcc aagtgtcgta ccggcatctc gtgctgctcc
     3361 acggcgtgtg catggctgga gacagcacca tggtgcagga atttgtacac ctgggggcca
     3421 tagacatgta tctgcgaaaa cgtggccacc tggtgccagc cagctggaag ctgcaggtgg
     3481 tcaaacagct ggcctacgcc ctcaactatc tggaggacaa aggcctgccc catggcaatg
     3541 tctctgcccg gaaggtgctc ctggctcggg agggggctga tgggagcccg cccttcatca
     3601 agctgagtga ccctggggtc agccccgctg tgttaagcct ggagatgctc accgacagga
     3661 tcccctgggt ggcccccgag tgtctccggg aggcgcagac acttagcttg gaagctgaca
     3721 agtggggctt cggcgccacg gtctgggaag tgtttagtgg cgtcaccatg cccatcagtg
     3781 ccctggatcc tgctaagaaa ctccaatttt atgaggaccg gcagcagctg ccggccccca
     3841 agtggacaga gctggccctg ctgattcaac agtgcatggc ctatgagccg gtccagaggc
     3901 cctccttccg agccgtcatt cgtgacctca atagcctcat ctcttcagac tatgagctcc
     3961 tctcagaccc cacacctggt gccctggcac ctcgtgatgg gctgtggaat ggtgcccagc
     4021 tctatgcctg ccaagacccc acgatcttcg aggagagaca cctcaagtac atctcacagc
     4081 tgggcaaggg caactttggc agcgtggagc tgtgccgcta tgacccgcta ggcgacaata
     4141 caggtgccct ggtggccgtg aaacagctgc agcacagcgg gccagaccag cagagggact
     4201 ttcagcggga gattcagatc ctcaaagcac tgcacagtga tttcattgtc aagtatcgtg
     4261 gtgtcagcta tggcccgggc cgccagagcc tgcggctggt catggagtac ctgcccagcg
     4321 gctgcttgcg cgacttcctg cagcggcacc gcgcgcgcct cgatgccagc cgcctccttc
     4381 tctattcctc gcagatctgc aagggcatgg agtacctggg ctcccgccgc tgcgtgcacc
     4441 gcgacctggc cgcccgaaac atcctcgtgg agagcgaggc acacgtcaag atcgctgact
     4501 tcggcctagc taagctgctg ccgcttgaca aagactacta cgtggtccgc gagccaggcc
     4561 agagccccat tttctggtat gcccccgaat ccctctcgga caacatcttc tctcgccagt
     4621 cagacgtctg gagcttcggg gtcgtcctgt acgagctctt cacctactgc gacaaaagct
     4681 gcagcccctc ggccgagttc ctgcggatga tgggatgtga gcgggatgtc cccgccctct
     4741 gccgcctctt ggaactgctg gaggagggcc agaggctgcc ggcgcctcct gcctgccctg
     4801 ctgaggttca cgagctcatg aagctgtgct gggcccctag cccacaggac cggccatcat
     4861 tcagcgccct gggcccccag ctggacatgc tgtggagcgg aagccggggg tgtgagactc
     4921 atgccttcac tgctcaccca gagggcaaac accactccct gtccttttca gtttaaacga
     4981 attcgggctc ggtacccggg gatcctctag agtcgacctg caggcatgca agctgatccg
     5041 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     5101 gcataacccc ttggggcggc cgcttcgagc agacatgata agatacattg atgagtttgg
     5161 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     5221 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     5281 ttttatgttt caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta
     5341 caaatgtggt aaaatcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta
     5401 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgcggatctg cgcagcacca
     5461 tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg cggaaagaac
     5521 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     5581 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     5641 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     5701 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     5761 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     5821 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttaa ttaactgttg
     5881 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
     5941 cccaggaggc agatcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     6001 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     6061 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     6121 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     6181 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     6241 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     6301 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     6361 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     6421 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     6481 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     6541 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     6601 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     6661 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     6721 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     6781 aagcgacgcc caacctgcca tcacgatggc cgcaataaaa tatctttatt ttcattacat
     6841 ctgtgtgttg gttttttgtg tgaatcgata gcgataagga tccgctcatg cgatgataag
     6901 ctgtcaaaca tgagaattac aacttatatc gtatggggct gacttcaggt gctacatttg
     6961 aagagataaa ttgcactgaa atctagaaat attttatctg attaataaga tgatcttctt
     7021 gagatcgttt tggtctgcgc gtaatctctt gctctgaaaa cgaaaaaacc gccttgcagg
     7081 gcggtttttc gaaggttctc tgagctacca actctttgaa ccgaggtaac tggcttggag
     7141 gagcgcagtc accaaaactt gtcctttcag tttagcctta accggcgcat gacttcaaga
     7201 ctaactcctc taaatcaatt accagtggct gctgccagtg gtgcttttgc atgtctttcc
     7261 gggttggact caagacgata gttaccggat aaggcgcagc ggtcggactg aacggggggt
     7321 tcgtgcatac agtccagctt ggagcgaact gcctacccgg aactgagtgt caggcgtgga
     7381 atgagacaaa cgcggccata acagcggaat gacaccggta aaccgaaagg caggaacagg
     7441 agagcgcacg agggagccgc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     7501 tcgccaccac tgatttgagc gtcagatttc gtgatgcttg tcaggggggc ggagcctatg
     7561 gaaaaacggc tttgccgcgg ccctctcact tccctgttaa gtatcttcct ggcatcttcc
     7621 aggaaatctc cgccccgttc gtaagccatt tccgctcgcc gcagtcgaac gaccgagcgt
     7681 agcgagtcag tgagcgagga agcggaatat atcctgtatc acatattctg ctgacgcacc
     7741 ggtgcagcct tttttctcct gccacatgaa gcacttcact gacaccctca tcagtgccaa
     7801 catagtaagc cagtatacac tccgctagcg ctgatgtccg gcggtgcttt tgccgttacg
     7861 caccaccccg tcatgagatt acgcgtttta gcggtgcgta caattaaggg attatggtaa
     7921 atccacttac tgtctgccct cgtagccatc gagataaacc gcagtactcc ggccacgatg
     7981 cgtccggcgt agaggatcga gatct
//
