LOCUS       sequence-471659-        6894 bp    DNA     circular SYN 06-MAY-2025
DEFINITION  Express FBXO42-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 6894)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6894)
  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: FBXO42-NanoLuc Vector
FEATURES             Location/Qualifiers
     source          1..6894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     61..81
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          246..378
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     361..380
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     422..441
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        422..440
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2626..3138
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(3226..3244)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3281..3402
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3292..3311)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3346..3365
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3647..4004
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3647..3838
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3670..3690)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3855..3990
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4019..4066
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4098..4892
                     /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(4152..4171)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4762..4781
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4956..5004
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5243..5831)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5323..5342)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5947..6230
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     enhancer        6421..6800
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(6801..6894,1..110)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga
       61 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
      121 accgtcagat cactagaagc tttattgcgg tagtttatca cagttaaatt gctaacgcag
      181 tcagtgcttc tgacacaaca gtctcgaact taagctgcag aagttggtcg tgaggcactg
      241 ggcaggtaag tatcaaggtt acaagacagg tttaaggaga ccaatagaaa ctgggcttgt
      301 cgagacagag aagactcttg cgtttctgat aggcacctat tggtcttact gacatccact
      361 ttgcctttct ctccacaggt gtccactccc agttcaatta cagctcttaa ggctagagta
      421 ttaatacgac tcactatagg gctagcgatc gccatggcca gctcctcgga cagtgaagat
      481 gacagtttca tggctgtgga ccaggaagaa actgtgctgg aagggacaat ggatcaagat
      541 gaggagcccc acccagtatt ggaggctgag gagactagac ataataggtc catgtcggag
      601 ctgccagaag aggttttgga gtatatcctg tcctttctct caccgtatca ggaacacaaa
      661 actgcggccc ttgtctgcaa acagtggtat cgacttatca aaggtgtagc ccatcagtgt
      721 tatcatggtt tcatgaaggc tgtccaggaa ggaaacattc agtgggagag ccgtacctat
      781 ccttatcctg gaaccccaat cactcagcgc ttctcgcaca gtgcatgcta ttatgatgct
      841 aatcagtcta tgtatgtgtt tggaggctgt acccagagca gctgcaatgc tgctttcaat
      901 gacctctgga gacttgacct aaacagcaaa gagtggatcc gacctttggc ttcagggtcc
      961 tatccttccc ccaaagctgg agcaactctg gtcgtgtaca aggacttgct agtgctgttt
     1021 ggtggctgga cgcggccaag cccttatccc ctacaccagc cagagagatt ctttgatgaa
     1081 atacacactt actcaccctc taaaaattgg tggaactgca ttgtgacaac ccatgggcca
     1141 cctcccatgg ctggccactc ctcctgtgtg atagatgata aaatgattgt ctttggtggc
     1201 tctttaggat cccggcaaat gagcaatgat gtctgggtcc ttgaccttga gcagtgggcg
     1261 tggtccaagc cgaacatctc tggccccagt cctcatcctc gaggtggcca atctcagatt
     1321 gtcatagatg atgcaactat cttaatcctc ggagggtgtg gcggtcccaa tgctctattc
     1381 aaggatgctt ggttgttgca catgcattct ggtccttggg cctggcagcc actcaaggta
     1441 gaaaatgaag agcatggggc cccagaactg tggtgccatc cagcttgccg ggtgggacag
     1501 tgtgtggtgg tcttcagcca ggctcctagt gggagagccc cactcagccc cagtttgaac
     1561 tctcgcccat cacctatcag tgccactcct ccagctctcg ttcctgaaac ccgagagtac
     1621 cgctctcagt ctccagtaag aagcatggat gaagctcctt gtgttaacgg ccgctgggga
     1681 acactgagac ccagggctca aaggcagact ccttcaggtt cccgggaagg gagcctttcc
     1741 ccagccagag gagacggctc tcctatcctc aatggtggga gtttgtctcc aggaacggca
     1801 gctgtgggtg gctcttcttt ggacagtcct gtacaggcca tatctccaag tactccatct
     1861 gctcctgaag gatacgacct gaaaatagga ctttctttgg ccccccgacg aggatcacta
     1921 ccagatcaga aagatctgag attaggatcc atagatctga attgggatct gaaacccgct
     1981 tccagtagta atcccatgga tggcatggac aataggacag ttgggggaag tatgagacac
     2041 cctcctgaac agacaaatgg tgtgcatacc ccacctcacg tggccagtgc ccttgcaggg
     2101 gccgtctccc caggtgccct gcgtcggagt ctggaagcca tcaaagcgat gtcctccaaa
     2161 ggcccctcgg cctctgcagc actaagtcct cctcttgggt cttctccagg ctctcctggg
     2221 agccagagtt tgagcagtgg agaaacagtg cccatccctc gcccagggcc tgcccaagga
     2281 gatggacatt ccttacctcc cattgctcgc cgcctgggcc accaccctcc acagtcccta
     2341 aatgttggca aacccctata ccagagtatg aactgcaagc ccatgcagat gtacgtgctg
     2401 gacattaaag acaccaagga gaaggggcgg gtcaaatgga aagtatttaa tagcagttct
     2461 gtggttggac ctcctgaaac cagcctgcat accgtggtac aaggcagggg tgaactcatc
     2521 atatttggag gactcatgga caagaaacag aatgtgaagt actatccaaa aacaaacgcc
     2581 ttgtactttg tacgagcaaa gagagtttct ctcggctcga gcggcgtctt cacactcgaa
     2641 gatttcgttg gggactggcg acagacagcc ggctacaacc tggaccaagt ccttgaacag
     2701 ggaggtgtgt ccagtttgtt tcagaatctc ggggtgtccg taactccgat ccaaaggatt
     2761 gtcctgagcg gtgaaaatgg gctgaagatc gacatccatg tcatcatccc gtatgaaggt
     2821 ctgagcggcg accaaatggg ccagatcgaa aaaattttta aggtggtgta ccctgtggat
     2881 gatcatcact ttaaggtgat cctgcactat ggcacactgg taatcgacgg ggttacgccg
     2941 aacatgatcg actatttcgg acggccgtat gaaggcatcg ccgtgttcga cggcaaaaag
     3001 atcactgtaa cagggaccct gtggaacggc aacaaaatta tcgacgagcg cctgatcaac
     3061 cccgacggct ccctgctgtt ccgagtaacc atcaacggag tgaccggctg gcggctgtgc
     3121 gaacgcattc tggcgtaagg ccgcgactct agagtcgacc tgcaggcatg caagctgatc
     3181 cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac
     3241 tagcataacc ccttggggcg gccgcttcga gcagacatga taagatacat tgatgagttt
     3301 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     3361 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     3421 cattttatgt ttcaggttca gggggagatg tgggaggttt ttttaagcaa gtaaaacctc
     3481 tacaaatgtg gtaaaatcga attttaacaa aatattaacg cttacaattt cctgatgcgg
     3541 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgcggatc tgcgcagcac
     3601 catggcctga aataacctct gaaagaggaa cttggttagg taccttctga ggcggaaaga
     3661 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3721 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3781 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3841 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3901 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     3961 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt aattaactgt
     4021 tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta
     4081 aacccaggag gcagatcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4141 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4201 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4261 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4321 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4381 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4441 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4501 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4561 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4621 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4681 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4741 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4801 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4861 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     4921 ccaagcgacg cccaacctgc catcacgatg gccgcaataa aatatcttta ttttcattac
     4981 atctgtgtgt tggttttttg tgtgaatcga tagcgataag gatcctcttt gcgcttgcgt
     5041 tttcccttgt ccagatagcc cagtagctga cattcatccg gggtcagcac cgtttctgcg
     5101 gactggcttt ctacccggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5161 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5221 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5281 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5341 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5401 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5461 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5521 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5581 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5641 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     5701 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5761 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5821 aaggatttca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5881 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5941 tatagtccgg aaatacagga acgcacgctg gatggccctt cgctgggatg gtgaaaccat
     6001 gaaaaatggc agcttcagtg gattaagtgg gggtaatgtg gcctgtaccc tctggttgca
     6061 taggtattca tacggttaaa atttatcagg cgcgattgcg gcagtttttc gggtggtttg
     6121 ttgccatttt tacctgtctg ctgccgtgat cgcgctgaac gcgttttagc ggtgcgtaca
     6181 attaagggat tatggtaaat ccacttactg tctgccctcg tagccatcga gataaaccgc
     6241 agtactccgg ccacgatgcg tccggcgtag aggatcgaga tcttcaatat tggccattag
     6301 ccatattatt cattggttat atagcataaa tcaatattgg ctattggcca ttgcatacgt
     6361 tgtatctata tcataatatg tacatttata ttggctcatg tccaatatga ccgccatgtt
     6421 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     6481 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     6541 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     6601 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     6661 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     6721 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
     6781 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
     6841 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtt
//
