LOCUS       sequence-473102-        6062 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of human ZRANB3 NZF (619-650).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6062)
  AUTHORS   Michel MA, Scutts S, Komander D
  TITLE     Secondary interactions in ubiquitin-binding domains achieve linkage 
            or substrate specificity.
  JOURNAL   Cell Rep. 2024 Aug 27;43(8):114545. doi: 
            10.1016/j.celrep.2024.114545. Epub 2024 Jul 23.
  PUBMED    39052481
REFERENCE   2  (bases 1 to 6062)
  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: "Cell Rep.";
            date: "2024-08-27"; volume: "43(8):114545. doi"; pages: " 
            10.1016/j.celrep.2024.114545. Epub 2024 Jul 23"
COMMENT     Sequence Label: pOPINK-ZRANB3 NZF (619-650)
FEATURES             Location/Qualifiers
     source          1..6062
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(109..156)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     152..170
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(223..240)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(360..383)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(402..1055)
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             complement(1056..1073)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1086..1108
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1123..1147
                     /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)."
     primer_bind     complement(1147..1166)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1148..1166)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1250..1269)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1475..1552
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1553..2635
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(1572..1591)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3444..3635
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3629..3651)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3737..3876
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3792..3811
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3891..3908)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4062..4650)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4142..4161)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4761..4780)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4772..5587
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             4772..5587
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4845..4864)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(join(5679..6062,1..72))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5816..5837)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6028..6047
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
       61 tacagggcgc gtcccattcg ccaatccgga tatagttcct cctttcagca aaaaacccct
      121 caagacccgt ttagaggccc caaggggtta tgctagttat tgctcagcgg tggcagcagc
      181 caactcagct tcctttcggg ctttgttagc agccggatct cagtggtggt ggtggtggtg
      241 tttaaatggt ctagaaagct ttaactgcct tgaggagtct cacacatttc acaataaggt
      301 aactctgaat tattgatata ggtgcagaga ctacattgcc agccctctac aggaaaggcc
      361 gggccctgaa acagaacttc cagaccgctg ctcagatccg attttggagg atggtcgcca
      421 ccaccaaacg tggcttgcca gccctgcaaa ggccatgcta tatacttgct ggatttcaag
      481 tacttatcaa tttgtgggat agcttcaata cgttttttaa aacaaactaa ttttgggaac
      541 gcatccaggc acattgggtc catgtataaa acaacatcaa gagcgtcata caacatgaag
      601 tcaggatggg ttacatgatc accatttaaa tatgttttat gacataaacg atcttcgaac
      661 attttcagca tttcaggtag cttgctaaga aaatcaactt tgagagtttc aaagtcttta
      721 ctatatgcaa ttctcgaaac accgtatcta atatccaaaa ccgctccttc aagcattgaa
      781 atctctgcac gctcttttgg acaaccaccc aacatgttgt gcttgtcagc tatataacgt
      841 atgatggcca tagactgtgt taatttaaca tcaccatcaa tataataagg aagattggga
      901 aactccaaac ccaattcaaa ctttttgttt cgccatttat caccttcatc gcgctcatac
      961 aaatgctctt catatttttc ttcaagatat tccaaaagaa gtcgagtggg ttgcacaagg
     1021 cccttaattt tccaataacc tagtataggg gacatgtgat ggtgatggtg atgtgccatg
     1081 gtatatctcc ttcttaaagt taaacaaaat tatttctaga ggggaattgt tatccgctca
     1141 caattcccct atagtgagtc gtattaattt cgcgggatcg agatctcgat cctctacgcc
     1201 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
     1261 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc
     1321 gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca
     1381 ccattccttg cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg
     1441 caggagtcgc ataagggaga gcgtcgagat cccggacacc atcgaatggc gcaaaacctt
     1501 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc
     1561 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt
     1621 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc
     1681 ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct
     1741 gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat
     1801 taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg
     1861 cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat
     1921 cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt
     1981 tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca
     2041 tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc
     2101 gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa
     2161 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat
     2221 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct
     2281 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg
     2341 cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat
     2401 cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg
     2461 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact
     2521 ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2581 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2641 acgcaattaa tgtaagttag ctcactcatt aggcaccggg atctcgaccg atgcccttga
     2701 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac
     2761 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca
     2821 ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat
     2881 tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg
     2941 gcgagaagca ggccattatc gccggcatgg cggccccacg ggtgcgcatg atcgtgctcc
     3001 tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag aatgaatcac
     3061 cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa
     3121 catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct
     3181 gcaccattat gttccggatc tgcatcgcag gatgctgctg gctaccctgt ggaacaccta
     3241 catctgtatt aacgaagcgc tggcattgac cctgagtgat ttttctctgg tcccgccgca
     3301 tccataccgc cagttgttta ccctcacaac gttccagtaa ccgggcatgt tcatcatcag
     3361 taacccgtat cgtgagcatc ctctctcgtt tcatcggtat cattaccccc atgaacagaa
     3421 atccccctta cacggaggca tcagtgacca aacaggaaaa aaccgccctt aacatggccc
     3481 gctttatcag aagccagaca ttaacgcttc tggagaaact caacgagctg gacgcggatg
     3541 aacaggcaga catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc
     3601 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     3661 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     3721 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg
     3781 gcttaactat gcggcatcag agcagattgt actgagagtg caccattgcg gtgtgaaata
     3841 ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact
     3901 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3961 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4021 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4081 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4141 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4201 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4261 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4321 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4381 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4441 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4501 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttgtt
     4561 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4621 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4681 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgaacaa taaaactgtc
     4741 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcttg
     4801 ctctaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     4861 cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc
     4921 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     4981 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     5041 tcctgatgat gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt
     5101 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     5161 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     5221 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     5281 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     5341 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     5401 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     5461 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     5521 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     5581 tttctaagaa ttaattcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5641 taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt
     5701 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     5761 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     5821 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     5881 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     5941 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     6001 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     6061 gc
//
