LOCUS       sequence-474363-        5582 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express CSNK1A1-NanoLuc(R) 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 5582)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5582)
  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            "Unpublished"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: NanoBRET Assay Vector, CSNK1A1-NanoLuc
FEATURES             Location/Qualifiers
     source          1..5582
                     /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             2061..2573
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2661..2679)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2716..2837
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2727..2746)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2781..2800
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3082..3439
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3082..3273
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3105..3125)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3290..3425
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3454..3501
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3533..4327
                     /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(3587..3606)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4197..4216
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4391..4439
                     /note="synthetic polyadenylation signal"
     rep_origin      4596..5141
                     /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     5042..5061
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5258..5275
                     /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 agtagcagcg
     1081 gctccaaggc tgaattcatt gtcggaggga aatataaact ggtacggaag atcgggtctg
     1141 gctccttcgg ggacatctat ttggcgatca acatcaccaa cggcgaggaa gtggcagtga
     1201 agctagaatc tcagaaggcc aggcatcccc agttgctgta cgagagcaag ctctataaga
     1261 ttcttcaagg tggggttggc atcccccaca tacggtggta tggtcaggaa aaagactaca
     1321 atgtactagt catggatctt ctgggaccta gcctcgaaga cctcttcaat ttctgttcaa
     1381 gaaggttcac aatgaaaact gtacttatgt tagctgacca gatgatcagt agaattgaat
     1441 atgtgcatac aaagaatttt atacacagag acattaaacc agataacttc ctaatgggta
     1501 ttgggcgtca ctgtaataag ttattcctta ttgattttgg tttggccaaa aagtacagag
     1561 acaacaggac aaggcaacac ataccataca gagaagataa aaacctcact ggcactgccc
     1621 gatatgctag catcaatgca catcttggta ttgagcagag tcgccgagat gacatggaat
     1681 cattaggata tgttttgatg tattttaata gaaccagcct gccatggcaa gggctaaagg
     1741 ctgcaacaaa gaaacaaaaa tatgaaaaga ttagtgaaaa gaagatgtcc acgcctgttg
     1801 aagttttatg taaggggttt cctgcagaat ttgcgatgta cttaaactat tgtcgtgggc
     1861 tacgctttga ggaagcccca gattacatgt atctgaggca gctattccgc attcttttca
     1921 ggaccctgaa ccatcaatat gactacacat ttgattggac aatgttaaag cagaaagcag
     1981 cacagcaggc agcctcttcc agtgggcagg gtcagcaggc ccaaaccccc acaggtttcg
     2041 tttctctcgg ctcgagcggc gtcttcacac tcgaagattt cgttggggac tggcgacaga
     2101 cagccggcta caacctggac caagtccttg aacagggagg tgtgtccagt ttgtttcaga
     2161 atctcggggt gtccgtaact ccgatccaaa ggattgtcct gagcggtgaa aatgggctga
     2221 agatcgacat ccatgtcatc atcccgtatg aaggtctgag cggcgaccaa atgggccaga
     2281 tcgaaaaaat ttttaaggtg gtgtaccctg tggatgatca tcactttaag gtgatcctgc
     2341 actatggcac actggtaatc gacggggtta cgccgaacat gatcgactat ttcggacggc
     2401 cgtatgaagg catcgccgtg ttcgacggca aaaagatcac tgtaacaggg accctgtgga
     2461 acggcaacaa aattatcgac gagcgcctga tcaaccccga cggctccctg ctgttccgag
     2521 taaccatcaa cggagtgacc ggctggcggc tgtgcgaacg cattctggcg taaggccgcg
     2581 actctagagt cgacctgcag gcatgcaagc tgatccggct gctaacaaag cccgaaagga
     2641 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcggccgc
     2701 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     2761 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     2821 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     2881 agatgtggga ggttttttta agcaagtaaa acctctacaa atgtggtaaa atcgaatttt
     2941 aacaaaatat taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg
     3001 gtatttcaca ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa cctctgaaag
     3061 aggaacttgg ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     3121 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3181 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3241 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3301 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3361 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     3421 aggcctaggc ttttgcaaaa agcttaatta actgttgaca attaatcatc ggcatagtat
     3481 atcggcatag tataatacga caaggtgagg aactaaaccc aggaggcaga tcatgattga
     3541 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     3601 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     3661 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     3721 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     3781 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     3841 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     3901 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     3961 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4021 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     4081 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4141 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4201 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4261 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     4321 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     4381 cgatggccgc aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga
     4441 atcgatagcg ataaggatcc gctcatgcga tgataagctg tcaaacatga gaattacaac
     4501 ttatatcgta tggggctgac ttcaggtgct acatttgaag agataaattg cactgaaatc
     4561 tagaaatatt ttatctgatt aataagatga tcttcttgag atcgttttgg tctgcgcgta
     4621 atctcttgct ctgaaaacga aaaaaccgcc ttgcagggcg gtttttcgaa ggttctctga
     4681 gctaccaact ctttgaaccg aggtaactgg cttggaggag cgcagtcacc aaaacttgtc
     4741 ctttcagttt agccttaacc ggcgcatgac ttcaagacta actcctctaa atcaattacc
     4801 agtggctgct gccagtggtg cttttgcatg tctttccggg ttggactcaa gacgatagtt
     4861 accggataag gcgcagcggt cggactgaac ggggggttcg tgcatacagt ccagcttgga
     4921 gcgaactgcc tacccggaac tgagtgtcag gcgtggaatg agacaaacgc ggccataaca
     4981 gcggaatgac accggtaaac cgaaaggcag gaacaggaga gcgcacgagg gagccgccag
     5041 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccaccactga tttgagcgtc
     5101 agatttcgtg atgcttgtca ggggggcgga gcctatggaa aaacggcttt gccgcggccc
     5161 tctcacttcc ctgttaagta tcttcctggc atcttccagg aaatctccgc cccgttcgta
     5221 agccatttcc gctcgccgca gtcgaacgac cgagcgtagc gagtcagtga gcgaggaagc
     5281 ggaatatatc ctgtatcaca tattctgctg acgcaccggt gcagcctttt ttctcctgcc
     5341 acatgaagca cttcactgac accctcatca gtgccaacat agtaagccag tatacactcc
     5401 gctagcgctg atgtccggcg gtgcttttgc cgttacgcac caccccgtca tgagattacg
     5461 cgttttagcg gtgcgtacaa ttaagggatt atggtaaatc cacttactgt ctgccctcgt
     5521 agccatcgag ataaaccgca gtactccggc cacgatgcgt ccggcgtaga ggatcgagat
     5581 ct
//
