LOCUS       sequence-81661-e        8347 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses human FbxO10 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 8347)
  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 8347)
  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-FbxO10-pcDNA3.1-
FEATURES             Location/Qualifiers
     source          1..8347
                     /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(3941..3958)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3947..4171
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4217..4645
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4304..4323)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4514..4535
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4654..4674)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4659..4988
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4839..4974
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4901..4920
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5055..5849
                     /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(5109..5128)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5719..5738
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6023..6144
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6060..6079)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6114..6133
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6193..6209)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6193..6209)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6206..6228)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6217..6233
                     /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(6241..6247,6248..6265,6266..6271))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6286..6307
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6424..6441)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6595..7180)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6675..6694)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7351..8142,8143..8211))
                     /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     7974..7993
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8212..8316)
                     /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 tagaatggag gctggtggcc tccccttgga
     1021 gctgtggcgc atgatcttag cctacttgca ccttcccgac ctgggccgct gcagcctggt
     1081 atgcagggcc tggtatgaac tgatcctcag tctcgacagc acccgctggc ggcagctgtg
     1141 tctgggttgc accgagtgcc gccatcccaa ttggcccaac cagccagatg tggagcctga
     1201 gtcttggaga gaagccttca agcagcatta ccttgcatcc aagacatgga ccaagaatgc
     1261 cttggacttg gagtcttcca tctgcttttc tctattccgc cggaggaggg aacgacgtac
     1321 cctgagtgtt gggccaggcc gtgagtttga cagcctgggc agtgccttgg ccatggccag
     1381 cctgtatgac cgaattgtgc tcttcccagg tgtgtacgaa gagcaaggtg aaatcatctt
     1441 gaaggtgcct gtggagattg tagggcaggg gaagttgggt gaagtggccc tgctggccag
     1501 cattgatcag cactgctcaa ccacacgcct gtgcaacctc gtcttcacgc cagcctggtt
     1561 ctcacccatc atgtataaga caacatcagg tcacgtccag tttgacaact gcaactttga
     1621 gaacgggcac atccaggtcc atggcccggg tacttgccaa gtgaagttct gtaccttcaa
     1681 aaacacccat atcttcctgc acaacgtgcc cctgtgtgtc ctggaaaact gtgaatttgt
     1741 gggcagtgaa aacaactctg tgactgttga gggtcaccca tctgcagata agaactgggc
     1801 ctacaagtat ctactagggc ttatcaagtc ctcacccact tttctcccca cagaggactc
     1861 tgacttttta atgtccctgg acctagagag ccgggaccag gcctggagcc caaagacctg
     1921 tgacattgtt atcgagggca gccagagccc taccagccca gcttctagct ccccaaagcc
     1981 aggctccaag gctggctcac aggaggcaga ggtgggtagt gatggtgaaa gggtggccca
     2041 gaccccggac agcagcgatg gaggcctgag tcccagcggt gaggatgaag atgaggacca
     2101 gctgatgtac agactatcct accaagtgca gggcccacgc cctgtattgg ggggctcatt
     2161 tctgggccca cctctaccag gagcatccat tcagctgccc agctgcctag tgctgaactc
     2221 actgcagcag gagctgcaga aggataagga ggccatggca ctggccaact ccgtgcaggg
     2281 ctgcctcatc cgcaagtgcc tcttccggga cgggaaggga ggcgtcttcg tctgctccca
     2341 cggcagagcc aagatggaag gaaacatctt ccggaacctg acttacgcag tgcggtgtat
     2401 acataatagc aagatcatca tgctcaggaa cgacatttac cgctgccgag cgtcaggcat
     2461 ctttcttcgc ttggagggcg gtggcttgat tgccggcaac aacatttacc acaatgcaga
     2521 ggctggtgta gacatccgga aaaagtccaa cccactcata ctgtgtaacc agatccacca
     2581 tggccttcgc tctggcattg tcgtccttgg caatgggaaa ggcatcatcc ggaacaatca
     2641 aatcttttcc aataaggagg ctggcattta catcctgtac cacggaaacc ccgttgtgag
     2701 cgggaaccac atcttcaagg gccgtgctgc cggcatagca gtgaatgaga acggcaaagg
     2761 cctcatcaca gaaaatgtca tccgtgagaa tcagtgggga ggtgtggaca tccgccgtgg
     2821 agggatcccc gttctcagga gtaacctcat ctgctttggc tattcagatg gtgtggttgt
     2881 gggagacgaa ggcaaaggcc tcatagaagg aaataccatc tacgctaaca agggctgtgg
     2941 tgtgtggatg atgtcgtcca gcctccccca tgtcaccagc aatcacgtca gctacaatgg
     3001 cctgtatgga gtggcagtat ttagccagaa ggatggctcc agcgagttac ctcgaggcca
     3061 cagggctcaa gagaacttca gcgaggatgg ggacgccatc ctctgggaga cagagctgga
     3121 gaaggaggac gacccactgc gccggcccat caccatagct cttgttgagt ctaacagtat
     3181 taatcacaat ggagcctcag gactctatgt ccagagcagc gaggcactgc atgtcatcac
     3241 caatgtgatc cacgcgaatg gggacagagg cattactgtg gcccagagca gccaacccac
     3301 ccgagtggcc aacaacagca tctcctgcaa ccggcaaagt ggggtcaagg ttgaggccca
     3361 gtgcaaagtg gagctccggg gcaatggtat ctatgacaac agaggccacg gcattatcac
     3421 caagggcgac agcaccatcg tcattgaaaa cgatatcatt ggcaaccggg gcagcgggct
     3481 gcagctgctg cccaggtccg acactaaagt aataaagaac cggatccact cgttccgggc
     3541 ctacggcatc gccgtgcggg gccgtgccaa ggccctggtg caggaaaaca tcatcttcca
     3601 gggcaaaacc agtaagacca tctttcagca gatctcaaac aaccgagaat gcatcatgca
     3661 aaacaacaag ttcctggtct tcaagaaaaa gtctgatacg tggcgcctgg tgaacccacc
     3721 agcacggccc caccttgaaa attctctcag acgtccctcg gcagcccaca atgggcagaa
     3781 ggtgacagcc atggcaacga ggatcacagc ccgggtggaa ggtggttacc acagcaaccg
     3841 cagtgtcttc tgcaccatcc tgtgagatct gcagaattcc accacactgg actagtggat
     3901 ccgagctcgg taccaagctt aagtttaaac cgctgatcag cctcgactgt gccttctagt
     3961 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     4021 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     4081 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     4141 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     4201 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4261 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4321 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4381 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4441 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4501 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4561 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4621 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     4681 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4741 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4801 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4861 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4921 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4981 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     5041 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     5101 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     5161 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     5221 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     5281 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     5341 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     5401 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     5461 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     5521 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     5581 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     5641 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5701 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5761 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5821 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     5881 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5941 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     6001 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     6061 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     6121 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     6181 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     6241 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     6301 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     6361 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     6421 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     6481 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6541 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6601 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6661 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6721 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6781 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6841 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6901 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6961 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     7021 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     7081 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt
     7141 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7201 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7261 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7321 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7381 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7441 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7501 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7561 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7621 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7681 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7741 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7801 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7861 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7921 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7981 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     8041 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8101 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8161 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8221 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8281 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8341 tgacgtc
//
