LOCUS       sequence-175029-        5828 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-tagged ubc-9, with a TEV protease cleavage site 
            between the tag and the ORF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5828)
  AUTHORS   Pelisch F, Sonneville R, Pourkarimi E, Agostinho A, Blow JJ, Gartner
            A, Hay RT
  TITLE     Dynamic SUMO modification regulates mitotic chromosome assembly and 
            cell cycle progression in Caenorhabditis elegans.
  JOURNAL   Nat Commun. 2014 Dec 5;5:5485. doi: 10.1038/ncomms6485.
  PUBMED    25475837
REFERENCE   2  (bases 1 to 5828)
  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: "Nat 
            Commun."; date: "2014-12-5"; volume: "5:5485. doi"; pages: " 
            10.1038/ncomms6485"
COMMENT     Sequence Label: 6xHis-TEV-UBC-9
FEATURES             Location/Qualifiers
     source          1..5828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      19..607
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     508..527
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     761..778
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    793..932
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(858..877)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1018..1040
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1034..1225)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2034..3116)
                     /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     3078..3097
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3117..3194)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3400..3419
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3507..3526
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3507..3525
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3526..3550
                     /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)."
     RBS             3565..3587
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3607..3624
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3646..3666
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             4195..4212
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4265..4283)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4279..4326
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4363..4818
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4450..4469)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4660..4681
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4910..5725)
                     /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             complement(4910..5725)
                     /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     5633..5652
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5717..5736
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
       61 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
      121 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
      181 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      241 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
      301 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
      361 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
      421 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
      481 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
      541 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
      601 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
      661 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
      721 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
      781 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
      841 caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
      901 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
      961 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     1021 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     1081 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     1141 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     1201 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg
     1261 gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac
     1321 atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc
     1381 agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac
     1441 agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact
     1501 tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg
     1561 tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct
     1621 gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca
     1681 tgcgcacccg tggggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg
     1741 tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat accgcaagcg
     1801 acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg acccagagcg
     1861 ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga
     1921 tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg
     1981 gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc
     2041 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     2101 ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc
     2161 aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg
     2221 gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag
     2281 ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg caccaacgcg cagcccggac
     2341 tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg
     2401 ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag
     2461 tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca
     2521 gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg
     2581 tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata
     2641 atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag
     2701 gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg
     2761 acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct
     2821 accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca
     2881 atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt
     2941 ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct
     3001 tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg
     3061 gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc
     3121 accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc
     3181 cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaagca
     3241 gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga
     3301 gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc
     3361 gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc
     3421 gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat
     3481 cgagatcgat ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga
     3541 taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat acatatgtcg
     3601 tactaccatc accatcacca tcacgattac gatatcccaa cgaccgaaaa cctgtatttt
     3661 cagggcgcca tgggatcggg aattgctgca ggacgcctcg cggaagaaag gaaacactgg
     3721 cgaaaggatc atccattcgg attcattgcc aaaccagtca agaacgccga cggaacattg
     3781 aacctcttca attgggaatg tgcaatccca gggagaaagg atacgatttg ggaaggcgga
     3841 ttatacagaa ttcggatgct cttcaaggac gatttcccgt caacgccacc aaagtgcaag
     3901 ttcgagccac cactcttcca tccaaatgtg tacccatcag gtaccgtgtg cttatctctt
     3961 ctggatgaaa acaaggattg gaagccgtca atctcaatca agcaacttct cattggaatt
     4021 caagatttgc tgaaccatcc aaatattgaa gatccagctc aggctgaagc ctatcagatc
     4081 tactgtcaga atagggctga atatgagaag cgagtgaaga aggaagctgt gaagtatgct
     4141 gccgaactcg tccagaagca aatgctcgag tagaagcttg cggccgcact cgagcaccac
     4201 caccaccacc actgagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
     4261 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
     4321 tttttgctga aaggaggaac tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg
     4381 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4441 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4501 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4561 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4621 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4681 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     4741 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     4801 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4861 tttatttttc taaatacatt caaatatgta tccgctcatg aattaattct tagaaaaact
     4921 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
     4981 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
     5041 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
     5101 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
     5161 agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct
     5221 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
     5281 gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     5341 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     5401 cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     5461 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
     5521 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     5581 catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
     5641 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctagagcaag
     5701 acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca
     5761 gttttattgt tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5821 cccgtaga
//
