LOCUS       sequence-479505-        5696 bp    DNA     circular SYN 23-JUL-2025
DEFINITION  ExpressNanoLuc-CDK2 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 5696)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5696)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 23, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoBRET NanoLuc-CDK2 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    36..84
                     /note="synthetic polyadenylation signal"
     CDS             complement(148..942)
                     /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(259..278)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     869..888
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(974..1021)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(1036..1393)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1050..1185
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        1202..1393
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     1350..1370
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    1638..1759
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1675..1694)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1729..1748
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1796..1814
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(2876..3388)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(3401..3420)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3402..3420)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3462..3481)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          3464..3596
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(3732..3935)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3761..3781)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3936..4315
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    4506..4789
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     rep_origin      4905..5493
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5394..5413
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgcaagcgca aagaggatcc ttatcgctat cgattcacac aaaaaaccaa cacacagatg
       61 taatgaaaat aaagatattt tattgcggcc atcgtgatgg caggttgggc gtcgcttggt
      121 cggtcatttc gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc
      181 gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc
      241 gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc
      301 cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt
      361 cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgcgcgcctt
      421 gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg
      481 atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg
      541 gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat
      601 ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc
      661 caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac
      721 gcccgtcgtg gccagccacg atagccgcgc tgcctcgtcc tgcagttcat tcagggcacc
      781 ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc
      841 ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca
      901 agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgatctgcc tcctgggttt
      961 agttcctcac cttgtcgtat tatactatgc cgatatacta tgccgatgat taattgtcaa
     1021 cagttaatta agctttttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg
     1081 gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc
     1141 atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg
     1201 cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     1261 gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct
     1321 gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc acagctggtt
     1381 ctttccgcct cagaaggtac ctaaccaagt tcctctttca gaggttattt caggccatgg
     1441 tgctgcgcag atccgcgtat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     1501 cgcatcagga aattgtaagc gttaatattt tgttaaaatt cgattttacc acatttgtag
     1561 aggttttact tgcttaaaaa aacctcccac atctccccct gaacctgaaa cataaaatga
     1621 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     1681 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     1741 aactcatcaa tgtatcttat catgtctgct cgaagcggcc gccccaaggg gttatgctag
     1801 ttattgctca gcggtggcag cagccaactc agcttccttt cgggctttgt tagcagccgg
     1861 atcagcttgc atgcctgcag gtcgactcta gaggatcccc gggtaccgag cccgaattcg
     1921 tttaaacgag tcgaagatgg ggtactggct tggtcacatc ctggaagaaa gggtgagcca
     1981 gggctgcctt ggccgaaatc cgcttgttag ggtcgtagtg cagcatttgc gataacaagc
     2041 tccgtccatc ttcatccagg ggaggtacaa ctttactaaa atcttgccgg gcccacttgg
     2101 ggaaacttgg cttgtaatca ggcatagaag taactcctgg ccacaccacc tcatctgggg
     2161 tccccagagt ccgaaagatc cggaagagct ggtcaatctc agaatctcca gggaataggg
     2221 cccggcgagt caccatctca gcaaagatgc agcccaggct ccagatgtcc acagctgtgg
     2281 aataatattt gcagcccagg aggatttcag gagctcggta ccacagggtc accacctcat
     2341 gggtgtaagt acgaacaggg actccaaaag ctctggctag tccaaagtct gctagcttga
     2401 tggccccctc tgtgttaata agcagattct gaggtttaag gtctcggtgg aggacccgat
     2461 gagaatggca gaaagctagg ccctggagca gctggaacag atagctcttg atgaggggaa
     2521 gaggaatgcc agtgagagca gaggcatcca tgaatttctt gagatcttgg tgcagaaatt
     2581 caaaaaccag gtagagttta ttttctgtgt gaatgacatc cagcagcttg acaatattag
     2641 gatggttaag ctccttaagc agagagatct ctcggatggc agtactgggc acaccctcag
     2701 tctcagtgtc caggcggatt ttcttaagcg ccaccacctc tcccgtcaac ttgtttctgg
     2761 ctttgtacac aactccgtac gtgccctctc cgatcttttc caccttttgg aagttctcca
     2821 tggcgatcgc tccaccgctg ctccctccac cgctcccgcc accacctgaa ccacccgcca
     2881 gaatgcgttc gcacagccgc cagccggtca ctccgttgat ggttactcgg aacagcaggg
     2941 agccgtcggg gttgatcagg cgctcgtcga taattttgtt gccgttccac agggtccctg
     3001 ttacagtgat ctttttgccg tcgaacacgg cgatgccttc atacggccgt ccgaaatagt
     3061 cgatcatgtt cggcgtaacc ccgtcgatta ccagtgtgcc atagtgcagg atcaccttaa
     3121 agtgatgatc atccacaggg tacaccacct taaaaatttt ttcgatctgg cccatttggt
     3181 cgccgctcag accttcatac gggatgatga catggatgtc gatcttcagc ccattttcac
     3241 cgctcaggac aatcctttgg atcggagtta cggacacccc gagattctga aacaaactgg
     3301 acacacctcc ctgttcaagg acttggtcca ggttgtagcc ggctgtctgt cgccagtccc
     3361 caacgaaatc ttcgagtgtg aagaccatgg tgagcgctag ccctatagtg agtcgtatta
     3421 atactctagc cttaagagct gtaattgaac tgggagtgga cacctgtgga gagaaaggca
     3481 aagtggatgt cagtaagacc aataggtgcc tatcagaaac gcaagagtct tctctgtctc
     3541 gacaagccca gtttctattg gtctccttaa acctgtcttg taaccttgat acttacctgc
     3601 ccagtgcctc acgaccaact tctgcagctt aagttcgaga ctgttgtgtc agaagcactg
     3661 actgcgttag caatttaact gtgataaact accgcaataa agcttctagt gatctgacgg
     3721 ttcactaaac cagctctgct tatatagacc tcccaccgta cacgcctacc gcccatttgc
     3781 gtcaacgggg cggggttatt acgacatttt ggaaagtccc gttgattttg gtgccaaaac
     3841 aaactcccat tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc aaaccgctat
     3901 ccacgcccat tggtgtactg ccaaaaccgc atcaccatgg taatagcgat gactaatacg
     3961 tagatgtact gccaagtagg aaagtcccgt aaggtcatgt actgggcata atgccaggcg
     4021 ggccatttac cgtcattgac gtcaataggg ggcggacttg gcatatgata cacttgatgt
     4081 actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct
     4141 attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc
     4201 ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta
     4261 tgaactaatg accccgtaat tgattactat taataactag tcaataatca atgccaacat
     4321 ggcggtcata ttggacatga gccaatataa atgtacatat tatgatatag atacaacgta
     4381 tgcaatggcc aatagccaat attgatttat gctatataac caatgaataa tatggctaat
     4441 ggccaatatt gaagatctcg atcctctacg ccggacgcat cgtggccgga gtactgcggt
     4501 ttatctcgat ggctacgagg gcagacagta agtggattta ccataatccc ttaattgtac
     4561 gcaccgctaa aacgcgttca gcgcgatcac ggcagcagac aggtaaaaat ggcaacaaac
     4621 cacccgaaaa actgccgcaa tcgcgcctga taaattttaa ccgtatgaat acctatgcaa
     4681 ccagagggta caggccacat tacccccact taatccactg aagctgccat ttttcatggt
     4741 ttcaccatcc cagcgaaggg ccatccagcg tgcgttcctg tatttccgga ctataaaagg
     4801 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     4861 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaaa tccttttttt
     4921 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4981 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5041 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5101 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5161 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5221 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5281 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5341 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5401 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5461 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5521 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     5581 gtggataacc gtattaccgc ctttgagtga gctgataccg ggtagaaagc cagtccgcag
     5641 aaacggtgct gaccccggat gaatgtcagc tactgggcta tctggacaag ggaaaa
//
