LOCUS       sequence-471658-        6907 bp    DNA     circular SYN 06-MAY-2025
DEFINITION  Express NanoLuc(R)-FBXO42 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 6907)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6907)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 6, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: NanoLuc-FBXO42 Vector
FEATURES             Location/Qualifiers
     source          1..6907
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1668..1686)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    1723..1844
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1734..1753)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1788..1807
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2089..2446
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2089..2280
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2112..2132)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2297..2432
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2527..3321
                     /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(2581..2600)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3191..3210
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3385..3433
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(3672..4260)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3752..3771)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4376..4659
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     enhancer        4850..5229
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5230..5433
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5384..5404
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          5569..5701
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     5684..5703
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     5745..5764
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5745..5763
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5777..6289
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
ORIGIN
        1 ctacaccagc cagagagatt ctttgatgaa atacacactt actcaccctc taaaaattgg
       61 tggaactgca ttgtgacaac ccatgggcca cctcccatgg ctggccactc ctcctgtgtg
      121 atagatgata aaatgattgt ctttggtggc tctttaggat cccggcaaat gagcaatgat
      181 gtctgggtcc ttgaccttga gcagtgggcg tggtccaagc cgaacatctc tggccccagt
      241 cctcatcctc gaggtggcca atctcagatt gtcatagatg atgcaactat cttaatcctc
      301 ggagggtgtg gcggtcccaa tgctctattc aaggatgctt ggttgttgca catgcattct
      361 ggtccttggg cctggcagcc actcaaggta gaaaatgaag agcatggggc cccagaactg
      421 tggtgccatc cagcttgccg ggtgggacag tgtgtggtgg tcttcagcca ggctcctagt
      481 gggagagccc cactcagccc cagtttgaac tctcgcccat cacctatcag tgccactcct
      541 ccagctctcg ttcctgaaac ccgagagtac cgctctcagt ctccagtaag aagcatggat
      601 gaagctcctt gtgttaacgg ccgctgggga acactgagac ccagggctca aaggcagact
      661 ccttcaggtt cccgggaagg gagcctttcc ccagccagag gagacggctc tcctatcctc
      721 aatggtggga gtttgtctcc aggaacggca gctgtgggtg gctcttcttt ggacagtcct
      781 gtacaggcca tatctccaag tactccatct gctcctgaag gatacgacct gaaaatagga
      841 ctttctttgg ccccccgacg aggatcacta ccagatcaga aagatctgag attaggatcc
      901 atagatctga attgggatct gaaacccgct tccagtagta atcccatgga tggcatggac
      961 aataggacag ttgggggaag tatgagacac cctcctgaac agacaaatgg tgtgcatacc
     1021 ccacctcacg tggccagtgc ccttgcaggg gccgtctccc caggtgccct gcgtcggagt
     1081 ctggaagcca tcaaagcgat gtcctccaaa ggcccctcgg cctctgcagc actaagtcct
     1141 cctcttgggt cttctccagg ctctcctggg agccagagtt tgagcagtgg agaaacagtg
     1201 cccatccctc gcccagggcc tgcccaagga gatggacatt ccttacctcc cattgctcgc
     1261 cgcctgggcc accaccctcc acagtcccta aatgttggca aacccctata ccagagtatg
     1321 aactgcaagc ccatgcagat gtacgtgctg gacattaaag acaccaagga gaaggggcgg
     1381 gtcaaatgga aagtatttaa tagcagttct gtggttggac ctcctgaaac cagcctgcat
     1441 accgtggtac aaggcagggg tgaactcatc atatttggag gactcatgga caagaaacag
     1501 aatgtgaagt actatccaaa aacaaacgcc ttgtactttg tacgagcaaa gagagtttaa
     1561 acgaattcgg gctcggtacc cggggatcct ctagagtcga cctgcaggca tgcaagctga
     1621 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
     1681 actagcataa ccccttgggg cggccgcttc gagcagacat gataagatac attgatgagt
     1741 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     1801 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     1861 ttcattttat gtttcaggtt cagggggaga tgtgggaggt ttttttaagc aagtaaaacc
     1921 tctacaaatg tggtaaaatc gaattttaac aaaatattaa cgcttacaat ttcctgatgc
     1981 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgcgga tctgcgcagc
     2041 accatggcct gaaataacct ctgaaagagg aacttggtta ggtaccttct gaggcggaaa
     2101 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     2161 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     2221 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     2281 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     2341 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     2401 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttaattaact
     2461 gttgacaatt aatcatcggc atagtataat acgacaaggt gaggaactaa acccaggagg
     2521 cagatcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     2581 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     2641 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     2701 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     2761 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     2821 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     2881 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     2941 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3001 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     3061 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     3121 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     3181 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     3241 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     3301 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     3361 ccaacctgcc atcacgatgg ccgcaataaa atatctttat tttcattaca tctgtgtgtt
     3421 ggttttttgt gtgaatcgat agcgataagg atcctctttg cgcttgcgtt ttcccttgtc
     3481 cagatagccc agtagctgac attcatccgg ggtcagcacc gtttctgcgg actggctttc
     3541 tacccggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3601 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3661 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3721 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3781 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3841 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3901 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3961 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4021 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4081 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     4141 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4201 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatttcaa
     4261 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4321 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt atagtccgga
     4381 aatacaggaa cgcacgctgg atggcccttc gctgggatgg tgaaaccatg aaaaatggca
     4441 gcttcagtgg attaagtggg ggtaatgtgg cctgtaccct ctggttgcat aggtattcat
     4501 acggttaaaa tttatcaggc gcgattgcgg cagtttttcg ggtggtttgt tgccattttt
     4561 acctgtctgc tgccgtgatc gcgctgaacg cgttttagcg gtgcgtacaa ttaagggatt
     4621 atggtaaatc cacttactgt ctgccctcgt agccatcgag ataaaccgca gtactccggc
     4681 cacgatgcgt ccggcgtaga ggatcgagat cttcaatatt ggccattagc catattattc
     4741 attggttata tagcataaat caatattggc tattggccat tgcatacgtt gtatctatat
     4801 cataatatgt acatttatat tggctcatgt ccaatatgac cgccatgttg gcattgatta
     4861 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     4921 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     4981 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     5041 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     5101 atgccaagtc cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     5161 cagtacatga ccttacggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     5221 attaccatgg tgatgcggtt ttggcagtac accaatgggc gtggatagcg gtttgactca
     5281 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     5341 caacgggact ttccaaaatg tcgtaataac cccgccccgt tgacgcaaat gggcggtagg
     5401 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcactaga
     5461 agctttattg cggtagttta tcacagttaa attgctaacg cagtcagtgc ttctgacaca
     5521 acagtctcga acttaagctg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag
     5581 gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc
     5641 ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca
     5701 ggtgtccact cccagttcaa ttacagctct taaggctaga gtattaatac gactcactat
     5761 agggctagcg ctcaccatgg tcttcacact cgaagatttc gttggggact ggcgacagac
     5821 agccggctac aacctggacc aagtccttga acagggaggt gtgtccagtt tgtttcagaa
     5881 tctcggggtg tccgtaactc cgatccaaag gattgtcctg agcggtgaaa atgggctgaa
     5941 gatcgacatc catgtcatca tcccgtatga aggtctgagc ggcgaccaaa tgggccagat
     6001 cgaaaaaatt tttaaggtgg tgtaccctgt ggatgatcat cactttaagg tgatcctgca
     6061 ctatggcaca ctggtaatcg acggggttac gccgaacatg atcgactatt tcggacggcc
     6121 gtatgaaggc atcgccgtgt tcgacggcaa aaagatcact gtaacaggga ccctgtggaa
     6181 cggcaacaaa attatcgacg agcgcctgat caaccccgac ggctccctgc tgttccgagt
     6241 aaccatcaac ggagtgaccg gctggcggct gtgcgaacgc attctggcgg gctcgagcgg
     6301 cgcgatcgcc atggccagct cctcggacag tgaagatgac agtttcatgg ctgtggacca
     6361 ggaagaaact gtgctggaag ggacaatgga tcaagatgag gagccccacc cagtattgga
     6421 ggctgaggag actagacata ataggtccat gtcggagctg ccagaagagg ttttggagta
     6481 tatcctgtcc tttctctcac cgtatcagga acacaaaact gcggcccttg tctgcaaaca
     6541 gtggtatcga cttatcaaag gtgtagccca tcagtgttat catggtttca tgaaggctgt
     6601 ccaggaagga aacattcagt gggagagccg tacctatcct tatcctggaa ccccaatcac
     6661 tcagcgcttc tcgcacagtg catgctatta tgatgctaat cagtctatgt atgtgtttgg
     6721 aggctgtacc cagagcagct gcaatgctgc tttcaatgac ctctggagac ttgacctaaa
     6781 cagcaaagag tggatccgac ctttggcttc agggtcctat ccttccccca aagctggagc
     6841 aactctggtc gtgtacaagg acttgctagt gctgtttggt ggctggacgc ggccaagccc
     6901 ttatccc
//
