LOCUS       sequence-81660-e        7977 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses human FbxO11-Jeff mutant 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 7977)
  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 7977)
  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-FbxO11-Jeff-pcDNA3.1-
FEATURES             Location/Qualifiers
     source          1..7977
                     /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(3571..3588)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3577..3801
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3847..4275
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3934..3953)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4144..4165
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4284..4304)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4289..4618
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4469..4604
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4531..4550
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4685..5479
                     /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(4739..4758)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5349..5368
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5653..5774
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5690..5709)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5744..5763
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5823..5839)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5823..5839)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5836..5858)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5847..5863
                     /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(5871..5877,5878..5895,5896..5901))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5916..5937
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6054..6071)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6225..6810)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6305..6324)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6981..7772,7773..7841))
                     /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     7604..7623
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7842..7946)
                     /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 tagaatggtt gcagaagaat caggtcctgg
     1021 tgcacaaaat agtccatacc aacttcgtag aaaaactctt ttgccgaaaa gaacagcgtg
     1081 tcccacaaag aacagtatgg agggcgcctc aacttcaact acagaaaact ttggtcatcg
     1141 tgcaaaacgt gcaagagtgt ctggaaaatc acaagatcta tcagcagcac ctgctgaaca
     1201 gtatcttcag gagaaactgc cagatgaagt ggttctaaaa atcttctctt acttgctgga
     1261 acaggatctt tgtagagcag cttgtgtatg taaacgcttc agtgaacttg ctaatgatcc
     1321 aattttgtgg aaacgattat atatggaagt atttgaatat actcgcccta tgatgcatcc
     1381 tgaacctgga aaattctacc agattaatcc agaagagtat gaacatccaa atccctggaa
     1441 agagagtttc cagcagttgt ataaaggtgc acatgtaaag ccaggatttg ctgaacattt
     1501 ctacagtaac cctgcaagat ataaaggaag agaaaatatg ttgtattatg atactattga
     1561 agatgccctt ggtggggtac aagaggctca ttttgatgga cttatctttg ttcattctgg
     1621 aatatatact gatgaatgga tatatattga atctccaatc accatgattg gtgcagcacc
     1681 tgggaaagtg gcagacaaag ttataattga aaacactaga gattcaacct tcgtttttat
     1741 ggaaggctct gaagatgctt atgttggata tatgacaata aggtttaacc ctgatgacaa
     1801 atctgcacaa caccacaatg cacaccactg cttagagatt acagtaaatt gtagccctat
     1861 tattgatcac tgtatcatcc gaagtacatg tacagttggt tctgcagtat gtgttagtgg
     1921 tcaaggagca tgtcccacca tcaagcactg taacatcagt gactgtgaaa atgttggact
     1981 atatataaca gatcatgcac agggaatata tgaggataat gaaatttcca ataatgcgtt
     2041 agctgggatt tgggttaaaa atcatggaaa cccaattatt agacggaatc atattcatca
     2101 tggacgtgat gttggtgtgt tcacatttga tcatggcatg ggttactttg aaagttgcaa
     2161 tatacacaga aataggatag caggctttga agtaaaagcc tatgctaacc ctacagtggt
     2221 tcgatgtgaa attcaccatg ggcagactgg aggaatatat gtccatgaaa aaggaagagg
     2281 acaattcata gagaataaaa tctatgcaaa caactttgca ggtgtatgga ttacctcaaa
     2341 tagtgaccca acaataaggg gaaattctat atttaatgga aatcaaggag gagtttacat
     2401 ctttggtgat ggacgaggcc ttattgaagg aaatgacatt tatggcaatg cattagcagg
     2461 aattctaatt aggacaaaca gttgtccaat tgttcggcat aacaaaattc atgatggcca
     2521 gcatggtggg atttatgtgc atgaaaaggg acaaggagta atagaagaga atgaagttta
     2581 tagtaacact ctagctggag tctgggtgac aactggcagc actccagtac tgagaagaaa
     2641 ccggatacac agtggcaagc aggttggtgt ttatttttat gacaatggac atggagtgct
     2701 agaagacaat gatatctata atcatatgta ttcaggggtt cagataagga ctggaagcaa
     2761 ccccaaaatt agacgcaaca aaatctgggg aggacagaat ggtggaattc tagtttataa
     2821 ttctggtcta ggctgtatag aagacaatga aatatttgac aatgcaatgg ctggagtctg
     2881 gattaagaca gatagtaatc ctacactaag aagaaataaa atccatgatg gaagagatgg
     2941 tggcatctgt atatttaatg ggggtcgagg tctccttgaa gaaaatgata ttttcaggaa
     3001 tgctcaagca ggtgttctca tcagcactaa tagtcatcca atcttaagga aaaacagaat
     3061 atttgatgga tttgccgcag gtattgaaat tacaaatcac gcaactgcaa cactagaagg
     3121 caatcagatt tttaacaacc ggtttggagg cttattttta gcatctggtg ttaatgtgac
     3181 aatgaaagat aacaaaataa tgaacaatca agatgccata gaaaaggctg ttagtagagg
     3241 ccaatgttta tataaaatat caagttatac cagctatccc atgcatgatt tctacagatg
     3301 tcatacttgt aacaccacag atcgaaatgc catatgtgtg aactgcatta agaagtgcca
     3361 tcagggacat gatgtagagt ttattagaca tgataggttt ttctgtgact gtggtgctgg
     3421 aacactgtct aatccttgta cattagctgg tgagcctaca catgatacag atacactata
     3481 tgactctgct ccacctatag aatctaatac attgcagcac aactgaggat ccgagctcgg
     3541 taccaagctt aagtttaaac cgctgatcag cctcgactgt gccttctagt tgccagccat
     3601 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     3661 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     3721 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     3781 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     3841 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     3901 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     3961 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4021 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     4081 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     4141 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     4201 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     4261 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     4321 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     4381 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4441 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4501 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     4561 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     4621 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     4681 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     4741 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     4801 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4861 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4921 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4981 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     5041 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     5101 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     5161 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     5221 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     5281 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     5341 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     5401 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     5461 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     5521 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     5581 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     5641 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5701 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5761 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     5821 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     5881 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5941 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     6001 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     6061 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     6121 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     6181 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6241 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6301 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6361 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     6421 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6481 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6541 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6601 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6661 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6721 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag
     6781 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6841 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6901 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6961 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     7021 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     7081 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7141 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7201 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     7261 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     7321 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7381 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7441 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7501 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7561 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     7621 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7681 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7741 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7801 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7861 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7921 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
