LOCUS       sequence-471727-        8158 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of N-terminal tagged FLAG-FBXL17 (H. sapiens, 
            codon-optimized) in human cell lines..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8158)
  AUTHORS   Cao S, Garcia SF, Shi H, James EI, Kito Y, Shi H, Mao H, Kaisari S, 
            Rona G, Deng S, Goldberg HV, Ponce J, Ueberheide B, Lignitto L, 
            Guttman M, Pagano M, Zheng N
  TITLE     Recognition of BACH1 quaternary structure degrons by two F-box 
            proteins under oxidative stress.
  JOURNAL   Cell. 2024 Oct 30:S0092-8674(24)01188-7. doi: 
            10.1016/j.cell.2024.10.012.
  PUBMED    39504958
REFERENCE   2  (bases 1 to 8158)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2024 
            Oct 30:S0092-8674(24)01188-7. doi: 10.1016/j.cell.2024.10.012."
COMMENT     Sequence Label: FLAG-FBXL17
FEATURES             Location/Qualifiers
     source          1..8158
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     50..69
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(129..145)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(129..145)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(142..164)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    153..169
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(177..183,184..201,202..207))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    222..243
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(360..377)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(531..1119)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(611..630)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1290..2081,2082..2150))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1913..1932
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2151..2255)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2330..2349)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2521..2900
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2901..3104
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3055..3075
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3101..3125
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      3216..3225
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3225..3248
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    5406..5994
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5459..5479)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5877..5888)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6015..6034)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6059..6107
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6309..6737
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6396..6415)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6606..6627
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6746..6766)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6751..7080
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6931..7066
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6993..7012
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             7147..7941
                     /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(7201..7220)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7811..7830
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    join(8117..8158,1..80)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(join(8154..8158,1..15))
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
       61 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
      121 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
      181 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
      241 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
      301 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
      361 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
      421 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
      481 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
      541 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
      601 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
      661 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
      721 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
      781 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
      841 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
      901 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
      961 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     1021 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     1081 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     1141 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     1201 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     1261 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     1321 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     1381 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     1441 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     1501 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     1561 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     1621 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     1681 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     1741 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     1801 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     1861 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     1921 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     1981 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     2041 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     2101 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     2161 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     2221 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     2281 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
     2341 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     2401 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     2461 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
     2521 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     2581 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     2641 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     2701 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     2761 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     2821 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     2881 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
     2941 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     3001 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     3061 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
     3121 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
     3181 ccagcctccg gactctagag gatcgaaccc ttgccgccac catggactac aaggacgacg
     3241 atgacaaagg ctctggcggc catctgctga gcaaagagcc cagaaacaga cccagccaga
     3301 aacggcctag atgctgcagc tggtgcagaa gaagaaggcc cctgctgcgg ctgcctagaa
     3361 gaacaccagc taaagtgcct ccacagcctg ccgctcctag aagcagagat tgcttcttca
     3421 gaggcccttg catgctgtgc ttcatcgtgc atagtcctgg cgctccagct cctgccggac
     3481 ctgaagaaga acctcctctg agccctccac ctagagatgg cgcttatgcc gctgcctcta
     3541 gctctcagca cctggccaga agatatgccg ctctggccgc tgaagattgt gccgccgctg
     3601 caagaagatt tctgctgtct agtgctgccg ctgctgctgc agccgctgca tctgcatctt
     3661 ctccagccag ctgttgcaaa gaactgggac ttgcagctgc tgccgcatgg gaacaacagg
     3721 gcagatctct gttcctggcc tctctgggcc ctgtcagatt tcttggacct cctgccgccg
     3781 tgcagctgtt tagaggacct acaccatctc cagccgaact gcccacacct cctgagatgg
     3841 tctgcaagag aaaaggcgca ggcgtgccag cctgtacacc ttgcaaacaa cctagatgtg
     3901 gcggcggagg ttgtggtggt ggcggaggtg gcggtggcgg aggcggacct gctggtggcg
     3961 gagcttctcc acctagacct ccagatgctg gatgttgtca ggcccctgaa cagcctcctc
     4021 aacctctgtg tcctccacca agcagcccta catctgaagg cgctcctact gaagctggcg
     4081 gagatgctgt tagagcaggc ggaacagctc ctctgtctgc ccagcaacag catgaatgtg
     4141 gcgacgccga ttgcagagag agccccgaga atccttgcga ctgtcacaga gagccacctc
     4201 cagagacacc cgacatcaat cagctgccac ctagcatcct gctgaagatc ttttccaacc
     4261 tgagcctgga cgagagatgc ctgtctgcca gcctcgtgtg caagtattgg agagatctgt
     4321 gcctggactt ccagttctgg aagcagctgg acctgagcag cagacagcaa gtgaccgacg
     4381 agctgctcga gaagatcgcc agcagaagcc agaacatcat cgagatcaac atcagcgact
     4441 gccggtccat gagcgacaat ggcgtgtgtg tgctggcctt caagtgtccc ggcctgctga
     4501 gatacaccgc ctacagatgc aagcagctga gcgacacctc tatcattgcc gtggcctctc
     4561 actgcccact gctgcagaaa gtgcacgtgg gcaaccagga caagctgacc gatgagggac
     4621 tgaaacagct gggcagcaag tgcagagagc tgaaggatat ccacttcggc cagtgctaca
     4681 agatcagcga cgagggcatg atcgtgatcg ccaagggatg cctgaagctg cagcggatct
     4741 acatgcaaga aaacaagctg gtcaccgacc agagcgtgaa ggcctttgcc gaacactgtc
     4801 ctgagctgca gtacgtgggc ttcatgggct gtagcgtgac aagcaaaggc gtgatccacc
     4861 tgaccaagct gagaaacctg tccagcctgg atctgcggca catcaccgag ctggacaacg
     4921 agacagtgat ggaaatcgtg aagcggtgca agaatctgag cagcctgaac ctgtgcctga
     4981 actggatcat caacgacaga tgcgtggaag tgattgccaa agagggccag aacctgaaag
     5041 aactgtacct ggtgtcctgc aagatcaccg actacgccct gatcgccatc ggcagataca
     5101 gcatgaccat cgaaaccgtg gacgtcggct ggtgtaaaga gatcacagac cagggcgcca
     5161 cactgatcgc ccagtctagc aagtctctga gatacctggg cctgatgaga tgcgacaaag
     5221 tgaacgaagt gaccgtggaa cagctggtcc agcagtaccc tcacatcacc tttagcaccg
     5281 tgctgcagga ctgcaagagg accctggaaa gggcctatca gatgggctgg acccctaata
     5341 tgagcgccgc cagctcttga aagggttcga tccctaccgg ttagtaatga gtttgatatc
     5401 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     5461 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     5521 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     5581 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     5641 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     5701 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     5761 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
     5821 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     5881 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     5941 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa
     6001 cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     6061 gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt
     6121 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     6181 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     6241 agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg
     6301 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     6361 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     6421 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     6481 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     6541 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     6601 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     6661 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     6721 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     6781 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     6841 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     6901 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     6961 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     7021 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     7081 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     7141 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     7201 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     7261 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     7321 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     7381 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     7441 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     7501 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     7561 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     7621 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     7681 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     7741 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     7801 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     7861 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     7921 cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac
     7981 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     8041 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     8101 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaat
//
