LOCUS       sequence-475293-        8687 bp    DNA     circular SYN 30-JUL-2025
DEFINITION  Express TNIK-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 8687)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8687)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 30, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: TNIK-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..8687
                     /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             5166..5678
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(5766..5784)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    5821..5942
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5832..5851)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5886..5905
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        6187..6544
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6187..6378
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6210..6230)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6395..6530
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6559..6606
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6638..7432
                     /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(6692..6711)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7302..7321
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7496..7544
                     /note="synthetic polyadenylation signal"
     rep_origin      7701..8246
                     /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     8147..8166
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8363..8380
                     /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 cgccatggcg agcgactccc
     1081 cggctcgaag cctggatgaa atagatctct cggctctgag ggaccctgca gggatctttg
     1141 aattggtgga acttgttgga aatggaacat acgggcaagt ttataagggt cgtcatgtca
     1201 aaacgggcca gcttgcagcc atcaaggtta tggatgtcac aggggatgaa gaggaagaaa
     1261 tcaaacaaga aattaacatg ttgaagaaat attctcatca ccggaatatt gctacatact
     1321 atggtgcttt tatcaaaaag aacccaccag gcatggatga ccaactttgg ttggtgatgg
     1381 agttttgtgg tgctggctct gtcaccgacc tgatcaagaa cacaaaaggt aacacgttga
     1441 aagaggagtg gattgcatac atctgcaggg aaatcttacg ggggctgagt cacctgcacc
     1501 agcataaagt gattcatcga gatattaaag ggcaaaatgt cttgctgact gaaaatgcag
     1561 aagttaaact agtggacttt ggagtcagtg ctcagcttga tcgaacagtg ggcaggagga
     1621 atactttcat tggaactccc tactggatgg caccagaagt tattgcctgt gatgaaaacc
     1681 cagatgccac atatgatttc aagagtgact tgtggtcttt gggtatcacc gccattgaaa
     1741 tggcagaagg tgctccccct ctctgtgaca tgcaccccat gagagctctc ttcctcatcc
     1801 cccggaatcc agcgcctcgg ctgaagtcta agaagtggtc aaaaaaattc cagtcattta
     1861 ttgagagctg cttggtaaag aatcacagcc agcgaccagc aacagaacaa ttgatgaagc
     1921 atccatttat acgagaccaa cctaatgagc gacaggtccg cattcaactc aaggaccata
     1981 ttgatagaac aaagaagaag cgaggagaaa aagatgagac agagtatgag tacagtggaa
     2041 gtgaggaaga agaggaggag aatgactcag gagagcccag ctccatcctg aatctgccag
     2101 gggagtcgac gctgcggagg gactttctga ggctgcagct ggccaacaag gagcgttctg
     2161 aggccctacg gaggcagcag ctggagcagc agcagcggga gaatgaggag cacaagcggc
     2221 agctgctggc cgagcgtcag aagcgcatcg aggagcagaa agagcagagg cggcggctgg
     2281 aggagcaaca aaggcgagag aaggagctgc ggaagcagca ggagagggag cagcgccggc
     2341 actatgagga gcagatgcgc cgggaggagg agaggaggcg tgcggagcat gaacaggaat
     2401 acatcaggcg acagttagag gaggagcaga gacagttaga gatcttgcag cagcagctac
     2461 tgcatgaaca agctctactt ctggaatata agcgcaaaca attggaagaa cagagacaag
     2521 cagaaagact gcagaggcag ctaaagcaag aaagagacta cttagtttcc cttcagcatc
     2581 agcggcagga gcagaggcct gtggagaaga agccactgta ccattacaaa gaaggaatga
     2641 gtcctagtga gaagccagca tgggccaagg aggtagaaga acggtcaagg ctcaaccggc
     2701 aaagttcccc tgccatgcct cacaaggttg ccaacaggat atctgacccc aacctgcccc
     2761 caaggtcgga gtccttcagc attagtggag ttcagcctgc tcgaacaccc cccatgctca
     2821 gaccagtcga tccccagatc ccacatctgg tagctgtaaa atcccaggga cctgccttga
     2881 ccgcctccca gtcagtgcac gagcagccca caaagggcct ctctgggttt caggaggctc
     2941 tgaacgtgac ctcccaccgc gtggagatgc cacgccagaa ctcagatccc acctcggaaa
     3001 atcctcctct ccccactcgc attgaaaagt ttgaccgaag ctcttggtta cgacaggaag
     3061 aagacattcc accaaaggtg cctcaaagaa caacttctat atccccagca ttagccagaa
     3121 agaattctcc tgggaatggt agtgctctgg gacccagact aggatctcaa cccatcagag
     3181 caagcaaccc tgatctccgg agaactgagc ccatcttgga gagccccttg cagaggacca
     3241 gcagtggcag ttcctccagc tccagcaccc ctagctccca gcccagctcc caaggaggct
     3301 cccagcctgg atcacaagca ggatccagtg aacgcaccag agttcgagcc aacagtaagt
     3361 cagaaggatc acctgtgctt ccccatgagc ctgccaaggt gaaaccagaa gaatccaggg
     3421 acattacccg gcccagtcga ccagctagct acaaaaaagc tatagatgag gatctgacgg
     3481 cattagccaa agaactaaga gaactccgga ttgaagaaac aaaccgccca atgaagaagg
     3541 tgactgatta ctcctcctcc agtgaggagt cagaaagtag cgaggaagag gaggaagatg
     3601 gagagagcga gacccatgat gggacagtgg ctgtcagcga catacccaga ctgataccaa
     3661 caggagctcc aggcagcaac gagcagtaca atgtgggaat ggtggggacg catgggctgg
     3721 agacctctca tgcggacagt ttcagcggca gtatttcaag agaaggaacc ttgatgatta
     3781 gagagacgtc tggagagaag aagcgatctg gccacagtga cagcaatggc tttgctggcc
     3841 acatcaacct ccctgacctg gtgcagcaga gccattctcc agctggaacc ccgactgagg
     3901 gactggggcg cgtctcaacc cattcccagg agatggactc tgggactgaa tatggcatgg
     3961 ggagcagcac caaagcctcc ttcaccccct ttgtggaccc cagagtatac cagacgtctc
     4021 ccactgatga agatgaagag gatgaggaat catcagccgc agctctgttt actagcgaac
     4081 ttcttaggca agaacaggcc aaactcaatg aagcaagaaa gatttcggtg gtaaatgtaa
     4141 acccaaccaa cattcggcct catagcgaca caccagaaat cagaaaatac aagaaacgat
     4201 tcaactcaga aatactttgt gcagctctgt ggggtgtaaa ccttctggtg gggactgaaa
     4261 atggcctgat gcttttggac cgaagtgggc aaggcaaagt ctataatctg atcaaccgga
     4321 ggcgatttca gcagatggat gtgctagagg gactgaatgt ccttgtgaca atttcaggaa
     4381 agaagaataa gctacgagtt tactatcttt catggttaag aaacagaata ctacataatg
     4441 acccagaagt agaaaagaaa caaggctgga tcactgttgg ggacttggaa ggctgtatac
     4501 attataaagt tgttaaatat gaaaggatca aatttttggt gattgcctta aagaatgctg
     4561 tggaaatata tgcttgggct cctaaaccgt atcataaatt catggcattt aagtcttttg
     4621 cagatctcca gcacaagcct ctgctagttg atctcacggt agaagaaggt caaagattaa
     4681 aggttatttt tggttcacac actggtttcc atgtaattga tgttgattca ggaaactctt
     4741 atgatatcta cataccatct catattcagg gcaatatcac tcctcatgct attgtcatct
     4801 tgcctaaaac agatggaatg gaaatgcttg tttgctatga ggatgagggg gtgtatgtaa
     4861 acacctatgg ccggataact aaggatgtgg tgctccaatg gggagaaatg cccacgtctg
     4921 tggcctacat tcattccaat cagataatgg gctggggcga gaaagctatt gagatccggt
     4981 cagtggaaac aggacatttg gatggagtat ttatgcataa gcgagctcaa aggttaaagt
     5041 ttctatgtga aagaaatgat aaggtatttt ttgcatccgt gcgatctgga ggaagtagcc
     5101 aagtgttttt catgaccctc aacagaaatt ccatgatgaa ctgggtttct ctcggctcga
     5161 gcggcgtctt cacactcgaa gatttcgttg gggactggcg acagacagcc ggctacaacc
     5221 tggaccaagt ccttgaacag ggaggtgtgt ccagtttgtt tcagaatctc ggggtgtccg
     5281 taactccgat ccaaaggatt gtcctgagcg gtgaaaatgg gctgaagatc gacatccatg
     5341 tcatcatccc gtatgaaggt ctgagcggcg accaaatggg ccagatcgaa aaaattttta
     5401 aggtggtgta ccctgtggat gatcatcact ttaaggtgat cctgcactat ggcacactgg
     5461 taatcgacgg ggttacgccg aacatgatcg actatttcgg acggccgtat gaaggcatcg
     5521 ccgtgttcga cggcaaaaag atcactgtaa cagggaccct gtggaacggc aacaaaatta
     5581 tcgacgagcg cctgatcaac cccgacggct ccctgctgtt ccgagtaacc atcaacggag
     5641 tgaccggctg gcggctgtgc gaacgcattc tggcgtaagg ccgcgactct agagtcgacc
     5701 tgcaggcatg caagctgatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     5761 tgccaccgct gagcaataac tagcataacc ccttggggcg gccgcttcga gcagacatga
     5821 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     5881 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     5941 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
     6001 ttttaagcaa gtaaaacctc tacaaatgtg gtaaaatcga attttaacaa aatattaacg
     6061 cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     6121 tacgcggatc tgcgcagcac catggcctga aataacctct gaaagaggaa cttggttagg
     6181 taccttctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     6241 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     6301 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     6361 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     6421 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     6481 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     6541 caaaaagctt aattaactgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     6601 tacgacaagg tgaggaacta aacccaggag gcagatcatg attgaacaag atggattgca
     6661 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     6721 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     6781 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     6841 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     6901 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     6961 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     7021 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     7081 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     7141 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     7201 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     7261 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     7321 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     7381 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     7441 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgatg gccgcaataa
     7501 aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagcgataag
     7561 gatccgctca tgcgatgata agctgtcaaa catgagaatt acaacttata tcgtatgggg
     7621 ctgacttcag gtgctacatt tgaagagata aattgcactg aaatctagaa atattttatc
     7681 tgattaataa gatgatcttc ttgagatcgt tttggtctgc gcgtaatctc ttgctctgaa
     7741 aacgaaaaaa ccgccttgca gggcggtttt tcgaaggttc tctgagctac caactctttg
     7801 aaccgaggta actggcttgg aggagcgcag tcaccaaaac ttgtcctttc agtttagcct
     7861 taaccggcgc atgacttcaa gactaactcc tctaaatcaa ttaccagtgg ctgctgccag
     7921 tggtgctttt gcatgtcttt ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     7981 gcggtcggac tgaacggggg gttcgtgcat acagtccagc ttggagcgaa ctgcctaccc
     8041 ggaactgagt gtcaggcgtg gaatgagaca aacgcggcca taacagcgga atgacaccgg
     8101 taaaccgaaa ggcaggaaca ggagagcgca cgagggagcc gccaggggga aacgcctggt
     8161 atctttatag tcctgtcggg tttcgccacc actgatttga gcgtcagatt tcgtgatgct
     8221 tgtcaggggg gcggagccta tggaaaaacg gctttgccgc ggccctctca cttccctgtt
     8281 aagtatcttc ctggcatctt ccaggaaatc tccgccccgt tcgtaagcca tttccgctcg
     8341 ccgcagtcga acgaccgagc gtagcgagtc agtgagcgag gaagcggaat atatcctgta
     8401 tcacatattc tgctgacgca ccggtgcagc cttttttctc ctgccacatg aagcacttca
     8461 ctgacaccct catcagtgcc aacatagtaa gccagtatac actccgctag cgctgatgtc
     8521 cggcggtgct tttgccgtta cgcaccaccc cgtcatgaga ttacgcgttt tagcggtgcg
     8581 tacaattaag ggattatggt aaatccactt actgtctgcc ctcgtagcca tcgagataaa
     8641 ccgcagtact ccggccacga tgcgtccggc gtagaggatc gagatct
//
