LOCUS       sequence-81465-e        7839 bp    DNA     circular SYN 14-APR-2025
DEFINITION  expresses human FbxO11-Delta-Fbox 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 7839)
  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 7839)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-Delta-Fbox-pcDNA3.1-
FEATURES             Location/Qualifiers
     source          1..7839
                     /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(3433..3450)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3439..3663
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3709..4137
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3796..3815)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4006..4027
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4146..4166)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4151..4480
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4331..4466
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4393..4412
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4547..5341
                     /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(4601..4620)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5211..5230
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5515..5636
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5552..5571)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5606..5625
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5685..5701)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5685..5701)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5698..5720)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5709..5725
                     /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(5733..5739,5740..5757,5758..5763))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5778..5799
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5916..5933)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6087..6672)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6167..6186)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6843..7634,7635..7703))
                     /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     7466..7485
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7704..7808)
                     /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 gtatatggaa gtatttgaat atactcgccc tatgatgcat cctgaacctg gaaaattcta
     1261 ccagattaat ccagaagagt atgaacatcc aaatccctgg aaagagagtt tccagcagtt
     1321 gtataaaggt gcacatgtaa agccaggatt tgctgaacat ttctacagta accctgcaag
     1381 atataaagga agagaaaata tgttgtatta tgatactatt gaagatgccc ttggtggggt
     1441 acaagaggct cattttgatg gacttatctt tgttcattct ggaatatata ctgatgaatg
     1501 gatatatatt gaatctccaa tcaccatgat tggtgcagca cctgggaaag tggcagacaa
     1561 agttataatt gaaaacacta gagattcaac cttcgttttt atggaaggct ctgaagatgc
     1621 ttatgttgga tatatgacaa taaggtttaa ccctgatgac aaatctgcac aacaccacaa
     1681 tgcacaccac tgcttagaga ttacagtaaa ttgtagccct attattgatc actgtatcat
     1741 ccgaagtaca tgtacagttg gttctgcagt atgtgttagt ggtcaaggag catgtcccac
     1801 catcaagcac tgtaacatca gtgactgtga aaatgttgga ctatatataa cagatcatgc
     1861 acagggaata tatgaggata atgaaatttc caataatgcg ttagctggga tttgggttaa
     1921 aaatcatgga aacccaatta ttagacggaa tcatattcat catggacgtg atgttggtgt
     1981 gttcacattt gatcatggca tgggttactt tgaaagttgc aatatacaca gaaataggat
     2041 agcaggcttt gaagtaaaag cctatgctaa ccctacagtg gttcgatgtg aaattcacca
     2101 tgggcagact ggaggaatat atgtccatga aaaaggaaga ggacaattca tagagaataa
     2161 aatctatgca aacaactttg caggtgtatg gattacctca aatagtgacc caacaataag
     2221 gggaaattct atatttaatg gaaatcaagg aggagtttac atctttggtg atggacgagg
     2281 ccttattgaa ggaaatgaca tttatggcaa tgcattagca ggaattcaaa ttaggacaaa
     2341 cagttgtcca attgttcggc ataacaaaat tcatgatggc cagcatggtg ggatttatgt
     2401 gcatgaaaag ggacaaggag taatagaaga gaatgaagtt tatagtaaca ctctagctgg
     2461 agtctgggtg acaactggca gcactccagt actgagaaga aaccggatac acagtggcaa
     2521 gcaggttggt gtttattttt atgacaatgg acatggagtg ctagaagaca atgatatcta
     2581 taatcatatg tattcagggg ttcagataag gactggaagc aaccccaaaa ttagacgcaa
     2641 caaaatctgg ggaggacaga atggtggaat tctagtttat aattctggtc taggctgtat
     2701 agaagacaat gaaatatttg acaatgcaat ggctggagtc tggattaaga cagatagtaa
     2761 tcctacacta agaagaaata aaatccatga tggaagagat ggtggcatct gtatatttaa
     2821 tgggggtcga ggtctccttg aagaaaatga tattttcagg aatgctcaag caggtgttct
     2881 catcagcact aatagtcatc caatcttaag gaaaaacaga atatttgatg gatttgccgc
     2941 aggtattgaa attacaaatc acgcaactgc aacactagaa ggcaatcaga tttttaacaa
     3001 ccggtttgga ggcttatttt tagcatctgg tgttaatgtg acaatgaaag ataacaaaat
     3061 aatgaacaat caagatgcca tagaaaaggc tgttagtaga ggccaatgtt tatataaaat
     3121 atcaagttat accagctatc ccatgcatga tttctacaga tgtcatactt gtaacaccac
     3181 agatcgaaat gccatatgtg tgaactgcat taagaagtgc catcagggac atgatgtaga
     3241 gtttattaga catgataggt ttttctgtga ctgtggtgct ggaacactgt ctaatccttg
     3301 tacattagct ggtgagccta cacatgatac agatacacta tatgactctg ctccacctat
     3361 agaatctaat acattgcagc acaactgagg atccgagctc ggtaccaagc ttaagtttaa
     3421 accgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     3481 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     3541 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     3601 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     3661 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     3721 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3781 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3841 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3901 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     3961 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4021 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     4081 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     4141 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     4201 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     4261 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     4321 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4381 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     4441 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     4501 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     4561 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4621 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4681 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     4741 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4801 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4861 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4921 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4981 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     5041 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     5101 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     5161 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     5221 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     5281 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     5341 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     5401 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5461 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     5521 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5581 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5641 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     5701 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     5761 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     5821 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5881 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5941 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6001 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6061 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6121 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6181 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6241 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     6301 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6361 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6421 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6481 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     6541 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6601 aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6661 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6721 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6781 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6841 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6901 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6961 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7021 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7081 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7141 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7201 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7261 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7321 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7381 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7441 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7501 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7561 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7621 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7681 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7741 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7801 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
