LOCUS       sequence-207918-        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        67..370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        371..574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     525..545
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        620..638
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      673..682
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             682..705
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3027..3046)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3028..3046)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3320..3441
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3357..3376)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3411..3430
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3448..3903)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3585..3606)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3797..3816
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3930..4034
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4036..4393
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4036..4227
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4059..4079)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4244..4379
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4428..5222
                     /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             4428..5222
                     /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(4482..4501)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5092..5111
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5410..5429)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5454..5501
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5830..6418
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6319..6338
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
       61 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      541 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      601 gcgattacgc caagctcgaa attaaccctc actaaaggga acaaaagctg gagctccacc
      661 gcggtggcgg ccgccaccat ggattacaag gatgacgacg ataagagccc gggcggatcc
      721 cccgggctgc aggaattgcc agaggaccaa gccggcgcag ccatggagga ggcgtctccc
      781 tattccttac ttgatatctg cttgaatttc ttgactactc accttgagaa gttctgttca
      841 gccagacaag atggaacatt gtgtctgcag gaacctggag tattcccaca ggaggtggct
      901 gatcgactgc ttcggaccat ggcttttcat ggtctattga atgatggaac tgtgggtatt
      961 tttaggggca accagatgcg cttaaagcga gcctgcattc gcaaagcaaa gatctctgct
     1021 gttgctttcc ggaaagcttt ctgccaccac aagttagtgg aacttgatgc cacaggtgtg
     1081 aatgctgata tcacgattac agacattatc agtgggcttg gcagtaacaa atggatccag
     1141 cagaatctcc agtgcctggt gctgaattca ttaactctct ccctcgagga tccttacgag
     1201 cgctgcttca gccggctttc tggccttcga gctttaagca tcacgaatgt tctcttttac
     1261 aatgaagacc tggctgaagt tgcctcattg ccaagattag agagcttgga tatttctaac
     1321 acctcaatca cagacatcac tgctctactg gcctgcaaag accgactcaa gtctctaacc
     1381 atgcaccact tgaaatgttt aaaaatgaca actacccaga tactggatgt agttcgggaa
     1441 ctcaaacatc tgaatcatct tgatatctca gatgataaac agtttacatc agacatagct
     1501 cttcgcttac tagaacaaaa agacatccta cctaaccttg tttctctgga tgtttctggg
     1561 agaaagcacg tgacagataa agccgttgaa gcctttatac aacaacgtcc aagcatgcaa
     1621 tttgtaggtt tgctggctac tgatgctggt tactctgaat tcctcacagg cgaaggacat
     1681 ttgaaggtgt ctggggaagc caatgaaact cagattgcag aagcactgaa gcgttacagt
     1741 gaacgggcat tctttgttcg ggaagctcta tttcatcttt ttagtctgac tcatgtgatg
     1801 gaaaaaacaa agccagaaat tttaaagctt gtggttactg ggatgagaaa ccaccctatg
     1861 aatttgccag tgcaactggc tgcaagcgcc tgtgtattta acttaaccaa gcaggatctt
     1921 gctgcaggga tgcctgtccg actcctggct gatgtgaccc atttgctgct caaagccatg
     1981 gaacattttc ccaatcacca gcagttacag aagaattgcc tcctttcact ttgcagtgac
     2041 cggatccttc aagatgttcc atttaacagg tttgaagcag ccaagcttgt catgcagtgg
     2101 ctttgcaacc atgaggatca aaacatgcaa aggatggcag ttgctatcat ttctatcctg
     2161 gctgccaagc tttctacaga acaaactgca cagcttggta ctgagctctt cattgtcagg
     2221 caacttcttc aaatagtgaa gcagaaaacc aatcaaaatt cagtggacac tacattgaaa
     2281 tttactttga gtgcactttg gaacctcaca gatgaatctc caaccacttg tagacacttt
     2341 attgaaaacc aagggttaga actcttcatg agggttctag agtctttccc aactgagtca
     2401 tccattcagc agaaagttct aggacttttg aacaatatag ctgaagtaca agaattacat
     2461 tctgaattaa tgtggaaaga ttttatagac cacatcagta gtctcctaca cagtgtggaa
     2521 gtggaagtca gttactttgc agctggaatt attgcccatt taatatccag aggtgaacaa
     2581 gcttggacat tgagtcgtag ccagaggaat tctctgctgg atgatttgca ttcagctatt
     2641 ttgaaatggc caactccaga gtgtgagatg gtagcataca ggtcctttaa tccatttttc
     2701 ccattacttg gctgtttcac aacaccagga gttcagctat gggcagtttg ggccatgcaa
     2761 catgtctgca gcaagaatcc ttcaaggtat tgcagcatgc tgattgaaga aggaggattg
     2821 cagcatttat acaacatcaa agatcatgaa catactgatc cccatgtcca acagattgct
     2881 gtggccattc tggatagctt agaaaaacac attgtgcgcc atgggaggcc acctccctgt
     2941 aaaaaacagc cccaagccag actaaattga tagtgagggc ccggtacctt aattaattaa
     3001 ggtaccaggt aagtgtaccc aattcgccct atagtgagtc gtattacaat tcactcgatc
     3061 gcccttccca acagttgcgc agcctgaatg gcgaatggag atccaatttt taagtgtata
     3121 atgtgttaaa ctactgattc taattgtttg tgtattttag attcacagtc ccaaggctca
     3181 tttcaggccc ctcagtcctc acagtctgtt catgatcata atcagccata ccacatttgt
     3241 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     3301 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     3361 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     3421 caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     3481 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     3541 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     3601 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     3661 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     3721 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     3781 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     3841 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     3901 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3961 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4021 gaaaaaggaa gaatcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     4081 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4141 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4201 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     4261 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     4321 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     4381 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     4441 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4501 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4561 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     4621 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4681 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4741 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4801 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4861 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagaa catcaggggc
     4921 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     4981 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     5041 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     5101 cccgtgatat tgctgaagaa cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     5161 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     5221 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5281 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5341 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     5401 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     5461 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     5521 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     5581 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     5641 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     5701 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5761 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5821 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5881 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5941 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     6001 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6061 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6121 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6181 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6241 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6301 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6361 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6421 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6481 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcc
//
