LOCUS       sequence-473350-        6015 bp    DNA     circular SYN 06-JUN-2025
DEFINITION  Express Skp2-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 6015)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6015)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 6, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: NanoBRET Assay Vector, Skp2
FEATURES             Location/Qualifiers
     source          1..6015
                     /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             2358..2870
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2958..2976)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3013..3134
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3024..3043)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3078..3097
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3379..3736
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3379..3570
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3402..3422)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3587..3722
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3751..3798
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3830..4624
                     /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(3884..3903)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4494..4513
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4688..4736
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4975..5563)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5055..5074)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5679..5962
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
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 cgccatgcac aggaagcacc
     1081 tccaggagat tccagacctg agtagcaacg ttgccaccag cttcacgtgg ggatgggatt
     1141 ccagcaagac ttctgaactg ctgtcaggca tgggggtctc cgccctggag aaagaggagc
     1201 ccgacagtga gaacatcccc caggaactgc tctcaaacct gggccacccg gagagccccc
     1261 cacggaaacg gctgaagagc aaagggagtg acaaagactt tgtgattgtc cgcaggccta
     1321 agctaaatcg agagaacttt ccaggtgttt catgggactc ccttccggat gagctgctct
     1381 tgggaatctt ttcctgtctg tgcctccctg agctgctaaa ggtctctggt gtttgtaaga
     1441 ggtggtatcg cctagcgtct gatgagtctc tatggcagac cttagacctc acaggtaaaa
     1501 atctgcaccc ggatgtgact ggtcggttgc tgtctcaagg ggtgattgcc ttccgctgcc
     1561 cacgatcatt tatggaccaa ccattggctg aacatttcag cccttttcgt gtacagcaca
     1621 tggacctatc gaactcagtt atagaagtgt ccaccctcca cggcatactg tctcagtgtt
     1681 ccaagttgca gaatctaagc ctggaaggcc tgcggctttc ggatcccatt gtcaatactc
     1741 tcgcaaaaaa ctcaaattta gtgcgactta acctttctgg gtgttctgga ttctctgaat
     1801 ttgccctgca gactttgcta agcagctgtt ccagactgga tgagctgaac ctctcctggt
     1861 gttttgattt cactgaaaag catgtacagg tggctgttgc gcatgtgtca gagaccatca
     1921 cccagctgaa tcttagcggc tacagaaaga atctccagaa atcagatctc tctactttag
     1981 ttagaagatg ccccaatctt gtccatctag acttaagtga tagtgtcatg ctaaagaatg
     2041 actgctttca ggaatttttc cagctcaact acctccaaca cctatcactc agtcggtgct
     2101 atgatataat acctgaaact ttacttgaac ttggagaaat tcccacacta aaaacactac
     2161 aagtttttgg aatcgtgcca gatggtaccc ttcaactgtt aaaggaagcc cttcctcatc
     2221 tacagattaa ttgctcccat ttcaccacca ttgccaggcc aactattggc aacaaaaaga
     2281 accaggagat atggggcatc aaatgccgac tgacactgca aaagcccagt tgtctagttt
     2341 ctctcggctc gagcggcgtc ttcacactcg aagatttcgt tggggactgg cgacagacag
     2401 ccggctacaa cctggaccaa gtccttgaac agggaggtgt gtccagtttg tttcagaatc
     2461 tcggggtgtc cgtaactccg atccaaagga ttgtcctgag cggtgaaaat gggctgaaga
     2521 tcgacatcca tgtcatcatc ccgtatgaag gtctgagcgg cgaccaaatg ggccagatcg
     2581 aaaaaatttt taaggtggtg taccctgtgg atgatcatca ctttaaggtg atcctgcact
     2641 atggcacact ggtaatcgac ggggttacgc cgaacatgat cgactatttc ggacggccgt
     2701 atgaaggcat cgccgtgttc gacggcaaaa agatcactgt aacagggacc ctgtggaacg
     2761 gcaacaaaat tatcgacgag cgcctgatca accccgacgg ctccctgctg ttccgagtaa
     2821 ccatcaacgg agtgaccggc tggcggctgt gcgaacgcat tctggcgtaa ggccgcgact
     2881 ctagagtcga cctgcaggca tgcaagctga tccggctgct aacaaagccc gaaaggaagc
     2941 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cggccgcttc
     3001 gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     3061 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     3121 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga
     3181 tgtgggaggt ttttttaagc aagtaaaacc tctacaaatg tggtaaaatc gaattttaac
     3241 aaaatattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta
     3301 tttcacaccg catacgcgga tctgcgcagc accatggcct gaaataacct ctgaaagagg
     3361 aacttggtta ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     3421 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3481 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3541 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3601 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3661 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3721 cctaggcttt tgcaaaaagc ttaattaact gttgacaatt aatcatcggc atagtatatc
     3781 ggcatagtat aatacgacaa ggtgaggaac taaacccagg aggcagatca tgattgaaca
     3841 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     3901 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     3961 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     4021 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     4081 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     4141 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     4201 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     4261 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     4321 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     4381 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4441 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4501 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     4561 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     4621 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     4681 tggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgaatc
     4741 gatagcgata aggatcctct ttgcgcttgc gttttccctt gtccagatag cccagtagct
     4801 gacattcatc cggggtcagc accgtttctg cggactggct ttctacccgg tatcagctca
     4861 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4921 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4981 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5041 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5101 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5161 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5221 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5281 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5341 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5401 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5461 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5521 tgtttgcaag cagcagatta cgcgcagaaa aaaaggattt caagaagatc ctttgatctt
     5581 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5641 attatcaaaa aggatcttca cctagatcct tttatagtcc ggaaatacag gaacgcacgc
     5701 tggatggccc ttcgctggga tggtgaaacc atgaaaaatg gcagcttcag tggattaagt
     5761 gggggtaatg tggcctgtac cctctggttg cataggtatt catacggtta aaatttatca
     5821 ggcgcgattg cggcagtttt tcgggtggtt tgttgccatt tttacctgtc tgctgccgtg
     5881 atcgcgctga acgcgtttta gcggtgcgta caattaaggg attatggtaa atccacttac
     5941 tgtctgccct cgtagccatc gagataaacc gcagtactcc ggccacgatg cgtccggcgt
     6001 agaggatcga gatct
//
