LOCUS       sequence-471847-        6245 bp    DNA     circular SYN 16-JUN-2025
DEFINITION  Express NanoLuc-SOS1cat in Mammalian Cells under a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6245)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6245)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: NanoBRET Assay Vector, NanoLuc-SOS1cat
FEATURES             Location/Qualifiers
     source          1..6245
                     /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(3188..3206)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3243..3364
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3254..3273)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3308..3327
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3609..3966
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3609..3800
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3632..3652)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3817..3952
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3981..4028
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4060..4854
                     /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(4114..4133)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4724..4743
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4918..4966
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5205..5793)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5285..5304)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5909..6192
                     /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 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 gctacaggaa gagaaagagg
     1621 agcagatgag gctgcctagt gctgatgttt atagatttgc agagcctgac tctgaagaga
     1681 atattatatt tgaagagaac atgcagccca aggctggaat tccaattatc aaagcaggaa
     1741 ctgttattaa acttatagag aggcttacgt accatatgta cgcagatccc aattttgttc
     1801 ggacatttct tacaacatac agatcctttt gcaaacctca agaactactg agtcttataa
     1861 tagaaaggtt tgaaattcca gagcctgagc caacagaagc tgatcgcata gctatagaga
     1921 atggagatca acccttgagt gcagaactga aaagatttag aaaagaatat atacagcctg
     1981 tgcaactgcg agtattaaat gtatgtcggc actgggtaga gcaccacttc tatgattttg
     2041 aaagagatgc atatcttttg caacgaatgg aagaatttat tggaacagta agaggtaaag
     2101 caatgaaaaa atgggttgaa tccatcacta aaataatcca aaggaaaaaa attgcaagag
     2161 acaatggacc aggtcataat attacatttc agagttcacc tcccacagtt gagtggcata
     2221 taagcagacc tgggcacata gagacttttg acctgctcac cttacaccca atagaaattg
     2281 ctcgacaact cactttactt gaatcagatc tataccgagc tgtacagcca tcagaattag
     2341 ttggaagtgt gtggacaaaa gaagacaaag aaattaactc tcctaatctt ctgaaaatga
     2401 ttcgacatac caccaacctc actctgtggt ttgagaaatg tattgtagaa actgaaaatt
     2461 tagaagaaag agtagctgtg gtgagtcgaa ttattgagat tctacaagtc tttcaagagt
     2521 tgaacaactt taatggtgtc cttgaggttg tcagtgctat gaattcatca cctgtttaca
     2581 gactagacca cacatttgag caaataccaa gtcgccagaa gaaaatttta gaagaagctc
     2641 atgaattgag tgaagatcac tataagaaat atttggcaaa actcaggtct attaatccac
     2701 catgtgtgcc tttctttgga atttatctca ctaatatctt gaaaacagaa gaaggcaacc
     2761 ctgaggtcct aaaaagacat ggaaaagagc ttataaactt tagcaaaagg aggaaagtag
     2821 cagaaataac aggagagatc cagcagtacc aaaatcagcc ttactgttta cgagtagaat
     2881 cagatatcaa aaggttcttt gaaaacttga atccgatggg aaatagcatg gagaaggaat
     2941 ttacagatta tcttttcaac aaatccctag aaatagaacc acgaaaccct aagcctctcc
     3001 caagatttcc aaaaaaatat agctatcccc taaaatctcc tggtgttcgt ccatcaaacc
     3061 caagaccagg taccgtttaa acgaattcgg gctcggtacc cggggatcct ctagagtcga
     3121 cctgcaggca tgcaagctga tccggctgct aacaaagccc gaaaggaagc tgagttggct
     3181 gctgccaccg ctgagcaata actagcataa ccccttgggg cggccgcttc gagcagacat
     3241 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     3301 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     3361 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt
     3421 ttttttaagc aagtaaaacc tctacaaatg tggtaaaatc gaattttaac aaaatattaa
     3481 cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     3541 catacgcgga tctgcgcagc accatggcct gaaataacct ctgaaagagg aacttggtta
     3601 ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     3661 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3721 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3781 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3841 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3901 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3961 tgcaaaaagc ttaattaact gttgacaatt aatcatcggc atagtatatc ggcatagtat
     4021 aatacgacaa ggtgaggaac taaacccagg aggcagatca tgattgaaca agatggattg
     4081 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4141 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4201 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     4261 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     4321 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4381 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4441 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     4501 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     4561 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     4621 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     4681 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     4741 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     4801 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     4861 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga tggccgcaat
     4921 aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgaatc gatagcgata
     4981 aggatcctct ttgcgcttgc gttttccctt gtccagatag cccagtagct gacattcatc
     5041 cggggtcagc accgtttctg cggactggct ttctacccgg tatcagctca ctcaaaggcg
     5101 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5161 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5221 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5281 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5341 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5401 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5461 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5521 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5581 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5641 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5701 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5761 cagcagatta cgcgcagaaa aaaaggattt caagaagatc ctttgatctt ttctacgggg
     5821 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5881 aggatcttca cctagatcct tttatagtcc ggaaatacag gaacgcacgc tggatggccc
     5941 ttcgctggga tggtgaaacc atgaaaaatg gcagcttcag tggattaagt gggggtaatg
     6001 tggcctgtac cctctggttg cataggtatt catacggtta aaatttatca ggcgcgattg
     6061 cggcagtttt tcgggtggtt tgttgccatt tttacctgtc tgctgccgtg atcgcgctga
     6121 acgcgtttta gcggtgcgta caattaaggg attatggtaa atccacttac tgtctgccct
     6181 cgtagccatc gagataaacc gcagtactcc ggccacgatg cgtccggcgt agaggatcga
     6241 gatct
//
