LOCUS       sequence-475317-        8029 bp    DNA     circular SYN 24-JUN-2025
DEFINITION  Express NanoLuc-JAK2 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 8029)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 8029)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 24, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: NanoLuc-JAK2 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..8029
                     /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(5108..5126)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    5163..5284
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5174..5193)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5228..5247
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5529..5886
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5529..5720
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5552..5572)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5737..5872
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5901..5948
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5980..6774
                     /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(6034..6053)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6644..6663
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6838..6886
                     /note="synthetic polyadenylation signal"
     rep_origin      7043..7588
                     /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     7489..7508
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7705..7722
                     /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 gggaatggcc tgccttacga
     1621 tgacagaaat ggagggaaca tccacctctt ctatatatca gaatggtgat atttctggaa
     1681 atgccaattc tatgaagcaa atagatccag ttcttcaggt gtatctttac cattcccttg
     1741 ggaaatctga ggcagattat ctgacctttc catctgggga gtatgttgca gaagaaatct
     1801 gtattgctgc ttctaaagct tgtggtatca cacctgtgta tcataatatg tttgctttaa
     1861 tgagtgaaac agaaaggatc tggtatccac ccaaccatgt cttccatata gatgagtcaa
     1921 ccaggcataa tgtactctac agaataagat tttactttcc tcgttggtat tgcagtggca
     1981 gcaacagagc ctatcggcat ggaatatctc gaggtgctga agctcctctt cttgatgact
     2041 ttgtcatgtc ttacctcttt gctcagtggc ggcatgattt tgtgcatgga tggataaaag
     2101 tacctgtgac tcatgaaaca caggaagaat gtcttgggat ggcagtgtta gatatgatga
     2161 gaatagccaa agaaaacgat caaaccccac tggccatcta taactctatc agctacaaga
     2221 cattcttacc aaaatgtatt cgagcaaaga tccaagacta tcatattttg acaaggaagc
     2281 gaataaggta cagatttcgc agatttattc agcaattcag ccaatgcaaa gccactgcca
     2341 gaaacttgaa acttaagtat cttataaatc tggaaactct gcagtctgcc ttctacacag
     2401 agaaatttga agtaaaagaa cctggaagtg gtccttcagg tgaggagatt tttgcaacca
     2461 ttataataac tggaaacggt ggaattcagt ggtcaagagg gaaacataaa gaaagtgaga
     2521 cactgacaga acaggattta cagttatatt gcgattttcc taatattatt gatgtcagta
     2581 ttaagcaagc aaaccaagag ggttcaaatg aaagccgagt tgtaactatc cataagcaag
     2641 atggtaaaaa tctggaaatt gaacttagct cattaaggga agctttgtct ttcgtgtcat
     2701 taattgatgg atattataga ttaactgcag atgcacatca ttacctctgt aaagaagtag
     2761 cacctccagc cgtgcttgaa aatatacaaa gcaactgtca tggcccaatt tcgatggatt
     2821 ttgccattag taaactgaag aaagcaggta atcagactgg actgtatgta cttcgatgca
     2881 gccctaagga ctttaataaa tattttttga cttttgctgt cgagcgagaa aatgtcattg
     2941 aatataaaca ctgtttgatt acaaaaaatg agaatgaaga gtacaacctc agtgggacaa
     3001 agaagaactt cagcagtctt aaagatcttt tgaattgtta ccagatggaa actgttcgct
     3061 cagacaatat aattttccag tttactaaat gctgtccccc aaagccaaaa gataaatcaa
     3121 accttctagt cttcagaacg aatggtgttt ctgatgtacc aacctcacca acattacaga
     3181 ggcctactca tatgaaccaa atggtgtttc acaaaatcag aaatgaagat ttgatattta
     3241 atgaaagcct tggccaaggc acttttacaa agatttttaa aggcgtacga agagaagtag
     3301 gagactacgg tcaactgcat gaaacagaag ttcttttaaa agttctggat aaagcacaca
     3361 gaaactattc agagtctttc tttgaagcag caagtatgat gagcaagctt tctcacaagc
     3421 atttggtttt aaattatgga gtatgtgtct gtggagacga gaatattctg gttcaggagt
     3481 ttgtaaaatt tggatcacta gatacatatc tgaaaaagaa taaaaattgt ataaatatat
     3541 tatggaaact tgaagttgct aaacagttgg catgggccat gcattttcta gaagaaaaca
     3601 cccttattca tgggaatgta tgtgccaaaa atattctgct tatcagagaa gaagacagga
     3661 agacaggaaa tcctcctttc atcaaactta gtgatcctgg cattagtatt acagttttgc
     3721 caaaggacat tcttcaggag agaataccat gggtaccacc tgaatgcatt gaaaatccta
     3781 aaaatttaaa tttggcaaca gacaaatgga gttttggtac cactttgtgg gaaatctgca
     3841 gtggaggaga taaacctcta agtgctctgg attctcaaag aaagctacaa ttttatgaag
     3901 ataggcatca gcttcctgca ccaaagtggg cagaattagc aaaccttata aataattgta
     3961 tggattatga accagatttc aggccttctt tcagagccat catacgagat cttaacagtt
     4021 tgtttactcc agattatgaa ctattaacag aaaatgacat gttaccaaat atgaggatag
     4081 gtgccctagg gttttctggt gcctttgaag accgggatcc tacacagttt gaagagagac
     4141 atttgaaatt tctacagcaa cttggcaagg gtaattttgg gagtgtggag atgtgccggt
     4201 atgaccctct acaggacaac actggggagg tggtcgctgt aaaaaagctt cagcatagta
     4261 ctgaagagca cctaagagac tttgaaaggg aaattgaaat cctgaaatcc ctacagcatg
     4321 acaacattgt aaagtacaag ggagtgtgct acagtgctgg tcggcgtaat ctaaaattaa
     4381 ttatggaata tttaccatat ggaagtttac gagactatct tcaaaaacat aaagaacgga
     4441 tagatcacat aaaacttctg cagtacacat ctcagatatg caagggtatg gagtatcttg
     4501 gtacaaaaag gtatatccac agggatctgg caacgagaaa tatattggtg gagaacgaga
     4561 acagagttaa aattggagat tttgggttaa ccaaagtctt gccacaagac aaagaatact
     4621 ataaagtaaa agaacctggt gaaagtccca tattctggta tgctccagaa tcactgacag
     4681 agagcaagtt ttctgtggcc tcagatgttt ggagctttgg agtggttctg tatgaacttt
     4741 tcacatacat tgagaagagt aaaagtccac cagcggaatt tatgcgtatg attggcaatg
     4801 acaaacaagg acagatgatc gtgttccatt tgatagaact tttgaagaat aatggaagat
     4861 taccaagacc agatggatgc ccagatgaga tctatatgat catgacagaa tgctggaaca
     4921 ataatgtaaa tcaacgcccc tcctttaggg atctagctct tcgagtggat caaataaggg
     4981 ataacatggc tggagtttaa acgaattcgg gctcggtacc cggggatcct ctagagtcga
     5041 cctgcaggca tgcaagctga tccggctgct aacaaagccc gaaaggaagc tgagttggct
     5101 gctgccaccg ctgagcaata actagcataa ccccttgggg cggccgcttc gagcagacat
     5161 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     5221 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     5281 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt
     5341 ttttttaagc aagtaaaacc tctacaaatg tggtaaaatc gaattttaac aaaatattaa
     5401 cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     5461 catacgcgga tctgcgcagc accatggcct gaaataacct ctgaaagagg aacttggtta
     5521 ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     5581 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     5641 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     5701 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     5761 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     5821 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     5881 tgcaaaaagc ttaattaact gttgacaatt aatcatcggc atagtatatc ggcatagtat
     5941 aatacgacaa ggtgaggaac taaacccagg aggcagatca tgattgaaca agatggattg
     6001 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     6061 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     6121 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     6181 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     6241 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     6301 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     6361 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     6421 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     6481 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     6541 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     6601 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     6661 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     6721 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     6781 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga tggccgcaat
     6841 aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgaatc gatagcgata
     6901 aggatccgct catgcgatga taagctgtca aacatgagaa ttacaactta tatcgtatgg
     6961 ggctgacttc aggtgctaca tttgaagaga taaattgcac tgaaatctag aaatatttta
     7021 tctgattaat aagatgatct tcttgagatc gttttggtct gcgcgtaatc tcttgctctg
     7081 aaaacgaaaa aaccgccttg cagggcggtt tttcgaaggt tctctgagct accaactctt
     7141 tgaaccgagg taactggctt ggaggagcgc agtcaccaaa acttgtcctt tcagtttagc
     7201 cttaaccggc gcatgacttc aagactaact cctctaaatc aattaccagt ggctgctgcc
     7261 agtggtgctt ttgcatgtct ttccgggttg gactcaagac gatagttacc ggataaggcg
     7321 cagcggtcgg actgaacggg gggttcgtgc atacagtcca gcttggagcg aactgcctac
     7381 ccggaactga gtgtcaggcg tggaatgaga caaacgcggc cataacagcg gaatgacacc
     7441 ggtaaaccga aaggcaggaa caggagagcg cacgagggag ccgccagggg gaaacgcctg
     7501 gtatctttat agtcctgtcg ggtttcgcca ccactgattt gagcgtcaga tttcgtgatg
     7561 cttgtcaggg gggcggagcc tatggaaaaa cggctttgcc gcggccctct cacttccctg
     7621 ttaagtatct tcctggcatc ttccaggaaa tctccgcccc gttcgtaagc catttccgct
     7681 cgccgcagtc gaacgaccga gcgtagcgag tcagtgagcg aggaagcgga atatatcctg
     7741 tatcacatat tctgctgacg caccggtgca gccttttttc tcctgccaca tgaagcactt
     7801 cactgacacc ctcatcagtg ccaacatagt aagccagtat acactccgct agcgctgatg
     7861 tccggcggtg cttttgccgt tacgcaccac cccgtcatga gattacgcgt tttagcggtg
     7921 cgtacaatta agggattatg gtaaatccac ttactgtctg ccctcgtagc catcgagata
     7981 aaccgcagta ctccggccac gatgcgtccg gcgtagagga tcgagatct
//
