LOCUS       sequence-81662-e        6840 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses human FbxO5 in mammalian cells with FLAG-HA tag at 
            N-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6840)
  AUTHORS   Horn M, Geisen C, Cermak L, Becker B, Nakamura S, Klein C, Pagano M,
            Antebi A
  TITLE     DRE-1/FBXO11-Dependent Degradation of BLMP-1/BLIMP-1 Governs C. 
            elegans Developmental Timing and Maturation.
  JOURNAL   Dev Cell. 2014 Mar 4. pii: S1534-5807(14)00069-0. doi: 
            10.1016/j.devcel.2014.01.028.
  PUBMED    24613396
REFERENCE   2  (bases 1 to 6840)
  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: "Dev Cell. 
            2014 Mar 4. pii: S1534-5807(14)00069-0. doi: 
            10.1016/j.devcel.2014.01.028."
COMMENT     Sequence Label: FLAG-HA-FbxO5-pcDNA3.1-
FEATURES             Location/Qualifiers
     source          1..6840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      899..908
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             908..931
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             947..973
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2434..2451)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2440..2664
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2710..3138
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2797..2816)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3007..3028
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3147..3167)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3152..3481
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3332..3467
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3394..3413
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3548..4342
                     /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(3602..3621)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4212..4231
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4516..4637
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4553..4572)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4607..4626
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4686..4702)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4686..4702)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4699..4721)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4710..4726
                     /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(4734..4740,4741..4758,4759..4764))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4779..4800
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4917..4934)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5088..5673)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5168..5187)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5844..6635,6636..6704))
                     /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     6467..6486
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6705..6809)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 caccatggac tacaaagacg atgacgataa agggggcggt ggaggttacc catacgatgt
      961 tccagattac gctggtggag gtggaggttc tagaatgagc cggcgcccct gcagctgcgc
     1021 cctacggcca ccccgctgct cctgcagcgc cagccccagc gcagtgacag ccgccgggcg
     1081 ccctcgaccc tcggatagtt gtaaagaaga aagttctacc ctttctgtca aaatgaagtg
     1141 tgattttaat tgtaaccatg ttcattccgg acttaaactg gtaaaacctg atgacattgg
     1201 aagactagtt tcctacaccc ctgcatattt ggaaggttcc tgtaaagact gcattaaaga
     1261 ctatgaaagg ctgtcatgta ttgggtcacc gattgtgagc cctaggattg tagaacttga
     1321 aactgaaagc aagcgcttgc ataacaagga aaatcaacat gtgcaacaga cacttaatag
     1381 tacaaatgaa atagaagcac tagagaccag tagactttat gaagacagtg gctattcctc
     1441 attttctcta caaagtggcc tcagtgaaca tgaagaaggt agcctcctgg aggagaattt
     1501 cggtgacagt ctacaatcct gcctgctaca aatacaaagc ccagaccaat atcccaacaa
     1561 aaacttgctg ccagttcttc attttgaaaa agtggtttgt tcaacattaa aaaagaatgc
     1621 aaaacgaaat cctaaagtag atcgggagat gctgaaggaa attatagcca gaggaaattt
     1681 tagactgcag aatataattg gcagaaaaat gggcctagaa tgtgtagata ttctcagcga
     1741 actctttcga aggggactca gacatgtctt agcaactatt ttagcacaac tcagtgacat
     1801 ggacttaatc aatgtgtcta aagtgagcac aacttggaag aagatcctag aagatgataa
     1861 gggggcattc cagttgtaca gtaaagcaat acaaagagtt accgaaaaca acaataaatt
     1921 ttcacctcat gcttcaacca gagaatatgt tatgttcaga accccactgg cttctgttca
     1981 gaaatcagca gcccagactt ctctcaaaaa agatgctcaa accaagttat ccaatcaagg
     2041 tgatcagaaa ggttctactt atagtcgaca caatgaattc tctgaggttg ccaagacatt
     2101 gaaaaagaac gaaagcctca aagcctgtat tcgctgtaat tcacctgcaa aatatgattg
     2161 ctatttacaa cgggcaacct gcaaacgaga aggctgtgga tttgattatt gtacgaagtg
     2221 tctctgtaat tatcatacta ctaaagactg ttcagatggc aagctcctca aagccagttg
     2281 taaaataggt cccctgcctg gtacaaagaa aagcaaaaag aatttacgaa gattgtgagc
     2341 ggccgccact gtgctggata tctgcagaat tccaccacac tggactagtg gatccgagct
     2401 cggtaccaag cttaagttta aaccgctgat cagcctcgac tgtgccttct agttgccagc
     2461 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     2521 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     2581 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     2641 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     2701 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     2761 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     2821 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     2881 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     2941 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3001 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     3061 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     3121 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3181 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3241 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3301 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3361 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3421 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3481 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     3541 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3601 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3661 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3721 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3781 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3841 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     3901 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     3961 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4021 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     4081 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4141 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4201 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4261 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4321 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4381 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4441 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4501 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     4561 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     4621 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     4681 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4741 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4801 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4861 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4921 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4981 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5041 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5101 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5161 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5221 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5281 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5341 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5401 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5461 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5521 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5581 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt tgtttgcaag
     5641 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5701 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5761 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5821 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5881 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5941 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6001 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6061 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6121 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6181 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6241 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6301 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6361 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6421 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6481 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6541 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6601 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6661 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6721 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6781 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
