LOCUS       sequence-211567-        7723 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses a FLAG-tagged human ZYG11A protein in mamalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7723)
  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 7723)
  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-ZYG11A
FEATURES             Location/Qualifiers
     source          1..7723
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        135..514
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        515..718
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     669..689
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     763..782
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        763..781
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      861..870
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             870..893
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3214..3231)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3220..3444
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3490..3918
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3577..3596)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3787..3808
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3927..3947)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3932..4261
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4112..4247
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4174..4193
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4328..5122
                     /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(4382..4401)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4992..5011
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5296..5417
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5333..5352)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5387..5406
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5466..5482)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5466..5482)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5479..5501)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5490..5506
                     /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(5514..5520,5521..5538,5539..5544))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5559..5580
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5697..5714)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5868..6456)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5948..5967)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6627..7418,7419..7487))
                     /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     7250..7269
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7488..7592)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7667..7686)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
       61 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
      121 cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
      181 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      241 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      301 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      361 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      421 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      481 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      541 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      601 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      661 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct
      721 ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag
      781 ggagacccaa gctggctagc gtttaaactt aagcttggta ccgagctcgg atccactagt
      841 ccagtgtggt ggaattcgcc gccaccatgg attacaagga tgacgacgat aagtctggcg
      901 ttcatttctt gcacccgggc cacacgcccc ggaacatcgt ccctcctgac gctcagaagg
      961 atgccctggg ctgctgcgtg gtacaggaag aggcatctcc ctacactttg gtcaacatct
     1021 gcctgaatgt cctgattgct aacctggaga aattgtgttc tgaaagacct gatggaacac
     1081 tgtgccttcc ggagcattgg agtttccctc aggaagtagc cgagcgattt ctcagggtga
     1141 tgacttggca aggcaagctg actgacagaa cagccagcat tttccgaggc aaccaaatga
     1201 aactgaagct ggtcaatatc caaaaagcta aaatctctac agctgcattc ataaaagcct
     1261 tctgccgtca taagctcatt gaactgaatg ctactgcagt gcacgctgac ctcccagttc
     1321 cagacatcat aagtggactc tgcagcaata ggtggatcca gcaaaacctc cagtgtctcc
     1381 tgttagactc gacaagcatc cctcaaaatt caagattact gttctttagt cagctcactg
     1441 gtcttcgcat tttaagtgtt tttaatgttt gttttcatac tgaagacctg gctaatgttt
     1501 ctcagttacc aagactggaa agcttagata tctctaatac tctagtcact gatatttctg
     1561 cactgcttac ctgtaaggat cgattgaagt ctctcacaat gcactatctg aaatgcctgg
     1621 ccatgaccaa atcacaaatt cttgcagtca ttagagaact taaatgtctg cttcaccttg
     1681 atatttctga tcacaggcaa ctcaaatcag acctagcttt tcatttgcta cagcagaagg
     1741 atatcctgcc caatgttgtg tcattggata tttctggggg caattgcatc actgatgaag
     1801 ctgtagaact gtttatacga ctgcggcctg ccatgcaatt tgtgggacta ttggccacgg
     1861 atgctggctc ttctgacttc tttactacaa agcaaggctt gagggttgct ggaggagcca
     1921 gtatgagtca gatttcagaa gcactgagcc gatacaggaa cagatcatgt tttgtgaagg
     1981 aagccctcca caggctgttc acagagacat tttcaatgga ggtaaccatg cctgctattt
     2041 taaagcttgt ggctatagga atgaggaatc acccattgga tttgcgagtg cagttcacag
     2101 ccagtgcttg cgctctcaac ctaacacgcc agggcctggc caaggggatg cctgttcgcc
     2161 tgttgtcaga ggtcacctgt ctacttttca aggctctgaa aaatttcccc cattaccagc
     2221 agttacagaa gaattgtctt ctctccttaa ccaattccag gattcttgtg gatgttccat
     2281 ttgacaggtt tgatgctgcc aagtttgtca tgagatggct ctgtaagcat gaaaacccca
     2341 agatgcaaac aatggcagtg agtgtcacct ctattctggc tctgcagctc tcacctgagc
     2401 aaacggcaca gcttgaagag cttttcatgg cagttaagga acttctagca atagtaaaac
     2461 aaaagactac tgagaattta gatgatgtca ccttcttgtt tactttgaaa gcactttgga
     2521 atcttacaga tgggtctcca gctgcctgca agcacttcat tgaaaatcaa ggattgcaaa
     2581 tcttcatcca agtcttggag accttttcag agtcagcaat acaaagcaaa gtacttggtc
     2641 ttttgaacaa catagcagaa gtcagagagc tctcttccaa gctggtgacc gaagatgtgc
     2701 tgaagcatat caacagttta ctctgtagca gggaaatgga agtcagctat tttgctgcag
     2761 gtatcatagc ccacctgaca tctgacagac agctttggat atcccgtgac ttccagaggc
     2821 gtactcttct ccaagatctg catgcaacca tacagaattg gccaagttca agttgtaaga
     2881 tgacagcatt ggtgacctat agatctttca agacattttt cccgctcctt ggcaacttct
     2941 ctcaaccaga ggttcagctc tgggcactat gggctatgta tcatgtctgc agtaaaaatc
     3001 ccagcaaata ctgcaaaatg ttagttgaag aagaaggatt gcagcttttg tgtgatatcc
     3061 aggagcacag tgaggcaacc cccaaagcac agcagattgc agcctccatt ctggatgact
     3121 tcagaatgca tttcatgaat tatcagaggc ccactctgtg tcaaatgccc ttctgactcg
     3181 agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     3241 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     3301 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     3361 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     3421 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     3481 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3541 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3601 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3661 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3721 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3781 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     3841 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     3901 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3961 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     4021 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4081 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4141 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4201 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4261 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     4321 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4381 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4441 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4501 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4561 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4621 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4681 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4741 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4801 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     4861 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4921 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4981 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     5041 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     5101 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     5161 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     5221 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     5281 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5341 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5401 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     5461 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5521 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5581 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5641 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5701 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5761 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5821 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5881 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5941 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6001 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6061 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6121 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6181 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6241 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6301 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6361 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6421 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6481 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6541 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6601 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6661 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6721 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6781 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6841 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6901 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6961 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7021 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7081 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7141 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7201 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7261 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7321 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7381 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7441 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7501 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7561 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7621 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
     7681 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gtt
//
