LOCUS       sequence-211568-        6526 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses a FLAG-tagged human ZYG11B protein in mamalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6526)
  AUTHORS   Balachandran RS, Heighington CS, Starostina NG, Anderson JW, Owen 
            DL, Vasudevan S, Kipreos ET
  TITLE     The ubiquitin ligase CRL2ZYG11 targets cyclin B1 for degradation in 
            a conserved pathway that facilitates mitotic slippage.
  JOURNAL   J Cell Biol. 2016 Oct 24;215(2):151-166. Epub 2016 Oct 17.
  PUBMED    27810909
REFERENCE   2  (bases 1 to 6526)
  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: "J Cell 
            Biol."; date: "2016-10-24"; volume: "215(2)"; pages: "151-166. Epub 
            2016 Oct 17"
COMMENT     Sequence Label: pFLAG-ZYG11B
FEATURES             Location/Qualifiers
     source          1..6526
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        86..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     544..564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        639..657
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      692..701
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             701..724
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3046..3065)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3047..3065)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3339..3460
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3376..3395)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3430..3449
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3467..3922)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3604..3625)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3816..3835
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3949..4053
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4055..4412
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4055..4246
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4078..4098)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4263..4398
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4447..5241
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4447..5241
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4501..4520)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5111..5130
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5429..5448)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5473..5520
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5849..6437
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6338..6357
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag
       61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      121 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      181 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      361 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
      601 tgaaccgtca gatccgctag cgattacgcc aagctcgaaa ttaaccctca ctaaagggaa
      661 caaaagctgg agctccaccg cggtggcggc cgccaccatg gattacaagg atgacgacga
      721 taagagcccg ggcggatccc ccgggctgca ggaattgcca gaggaccaag ccggcgcagc
      781 catggaggag gcgtctccct attccttact tgatatctgc ttgaatttct tgactactca
      841 ccttgagaag ttctgttcag ccagacaaga tggaacattg tgtctgcagg aacctggagt
      901 attcccacag gaggtggctg atcgactgct tcggaccatg gcttttcatg gtctattgaa
      961 tgatggaact gtgggtattt ttaggggcaa ccagatgcgc ttaaagcgag cctgcattcg
     1021 caaagcaaag atctctgctg ttgctttccg gaaagctttc tgccaccaca agttagtgga
     1081 acttgatgcc acaggtgtga atgctgatat cacgattaca gacattatca gtgggcttgg
     1141 cagtaacaaa tggatccagc agaatctcca gtgcctggtg ctgaattcat taactctctc
     1201 cctcgaggat ccttacgagc gctgcttcag ccggctttct ggccttcgag ctttaagcat
     1261 cacgaatgtt ctcttttaca atgaagacct ggctgaagtt gcctcattgc caagattaga
     1321 gagcttggat atttctaaca cctcaatcac agacatcact gctctactgg cctgcaaaga
     1381 ccgactcaag tctctaacca tgcaccactt gaaatgttta aaaatgacaa ctacccagat
     1441 actggatgta gttcgggaac tcaaacatct gaatcatctt gatatctcag atgataaaca
     1501 gtttacatca gacatagctc ttcgcttact agaacaaaaa gacatcctac ctaaccttgt
     1561 ttctctggat gtttctggga gaaagcacgt gacagataaa gccgttgaag cctttataca
     1621 acaacgtcca agcatgcaat ttgtaggttt gctggctact gatgctggtt actctgaatt
     1681 cctcacaggc gaaggacatt tgaaggtgtc tggggaagcc aatgaaactc agattgcaga
     1741 agcactgaag cgttacagtg aacgggcatt ctttgttcgg gaagctctat ttcatctttt
     1801 tagtctgact catgtgatgg aaaaaacaaa gccagaaatt ttaaagcttg tggttactgg
     1861 gatgagaaac caccctatga atttgccagt gcaactggct gcaagcgcct gtgtatttaa
     1921 cttaaccaag caggatcttg ctgcaggaat gcctgtccga ctcctggctg atgtgaccca
     1981 tttgctgctc aaagccatgg aacattttcc caatcaccag cagttacaga agaattgcct
     2041 cctttcactt tgcagtgacc ggatccttca agatgttcca tttaacaggt ttgaagcagc
     2101 caagcttgtc atgcagtggc tttgcaacca tgaggatcaa aacatgcaaa ggatggcagt
     2161 tgctatcatt tctatcctgg ctgccaagct ttctacagaa caaactgcac agcttggtac
     2221 tgagctcttc attgtcaggc aacttcttca aatagtgaag cagaaaacca atcaaaattc
     2281 agtggacact acattgaaat ttactttgag tgcactttgg aacctcacag atgaatctcc
     2341 aaccacttgt agacacttta ttgaaaacca agggttagaa ctcttcatga gggttctaga
     2401 gtctttccca actgagtcat ccattcagca gaaagttcta ggacttttga acaatatagc
     2461 tgaagtacaa gaattacatt ctgaattaat gtggaaagat tttatagacc acatcagtag
     2521 tctcctacac agtgtggaag tggaagtcag ttactttgca gctggaatta ttgcccattt
     2581 aatatccaga ggtgaacaag cttggacatt gagtcgtagc cagaggaatt ctctgctgga
     2641 tgatttgcat tcagctattt tgaaatggcc aactccagag tgtgagatgg tagcatacag
     2701 gtcctttaat ccatttttcc cattacttgg ctgtttcaca acaccaggag ttcagctatg
     2761 ggcagtttgg gccatgcaac atgtctgcag caagaatcct tcaaggtatt gcagcatgct
     2821 gattgaagaa ggaggattgc agcatttata caacatcaaa gatcatgaac atactgatcc
     2881 ccatgtccaa cagattgctg tggccattct ggatagctta gaaaaacaca ttgtgcgcca
     2941 tgggaggcca cctccctgta aaaaacagcc ccaagccaga ctaaattgat agtgagggcc
     3001 cggtacctta attaattaag gtaccaggta agtgtaccca attcgcccta tagtgagtcg
     3061 tattacaatt cactcgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggaga
     3121 tccaattttt aagtgtataa tgtgttaaac tactgattct aattgtttgt gtattttaga
     3181 ttcacagtcc caaggctcat ttcaggcccc tcagtcctca cagtctgttc atgatcataa
     3241 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     3301 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     3361 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3421 attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt
     3481 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     3541 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     3601 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     3661 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     3721 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     3781 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     3841 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     3901 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     3961 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4021 taaatgcttc aataatattg aaaaaggaag aatcctgagg cggaaagaac cagctgtgga
     4081 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4141 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4201 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4261 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4321 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     4381 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     4441 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4501 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4561 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4621 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4681 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4741 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4801 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4861 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4921 gacgaagaac atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     4981 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5041 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5101 caggacatag cgttggctac ccgtgatatt gctgaagaac ttggcggcga atgggctgac
     5161 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5221 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5281 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5341 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5401 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     5461 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     5521 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     5581 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     5641 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     5701 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5761 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5821 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5881 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5941 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6001 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6061 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6121 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6181 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6241 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6301 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6361 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6421 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6481 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgt
//
