LOCUS       sequence-175030-        6484 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-tagged gei-17 (isoform f), 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 6484)
  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 6484)
  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-GEI-17(133-509)
FEATURES             Location/Qualifiers
     source          1..6484
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      29..617
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     518..537
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     771..788
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    803..942
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(868..887)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1028..1050
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1044..1235)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2044..3126)
                     /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     3088..3107
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3127..3204)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3410..3429
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3517..3536
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3517..3535
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3536..3560
                     /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             3575..3597
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3617..3634
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3656..3676
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             4861..4878
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4931..4949)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4945..4992
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      5029..5484
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5116..5135)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5326..5347
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5576..6391)
                     /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(5576..6391)
                     /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     6299..6318
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6383..6402
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
       61 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
      121 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
      181 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
      241 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
      301 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
      361 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
      421 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
      481 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
      541 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
      601 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
      661 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
      721 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
      781 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
      841 tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
      901 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
      961 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     1021 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     1081 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     1141 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     1201 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     1261 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
     1321 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
     1381 cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta
     1441 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag
     1501 ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt
     1561 gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt
     1621 gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg
     1681 agcacgatca tgcgcacccg tggggccgcc atgccggcga taatggcctg cttctcgccg
     1741 aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat
     1801 accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg
     1861 acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt
     1921 gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
     1981 aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg
     2041 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     2101 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
     2161 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
     2221 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
     2281 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg caccaacgcg
     2341 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
     2401 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     2461 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     2521 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
     2581 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
     2641 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
     2701 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
     2761 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
     2821 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
     2881 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
     2941 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     3001 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
     3061 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
     3121 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
     3181 ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca
     3241 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg
     3301 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     3361 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     3421 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     3481 cgtagaggat cgagatcgat ctcgatcccg cgaaattaat acgactcact ataggggaat
     3541 tgtgagcgga taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat
     3601 acatatgtcg tactaccatc accatcacca tcacgattac gatatcccaa cgaccgaaaa
     3661 cctgtatttt cagggcgccg gtggcggtag ccaacaaatg atggctagtc atcacagtca
     3721 tttgcagcaa cagcaccatc ctagcacccc aaagaaaatg tacgccgaca actttgagcc
     3781 tttgccactt cccttttacg acgttatctc ggtcttgtta aaacctgtag agttgcattc
     3841 gtccgactcg ccgaccctta aacaaacgaa acagttacag tttccatttc tgctgacagc
     3901 ggaacatatt tccaaaatct cttatcgcgc cgatgtcaca cctttgccgc gctacgagtt
     3961 acagcttcgc tttttcaact taacggaacc ggtccaagga ccgcaaaagg atgattttcc
     4021 gctgaactgt tacgcccgtg tcgacgactc tgtagttcag ttgcccaatg taattcccac
     4081 aaacaaaaca aacgcagagc ccaagcgtcc atcccgtcca gtcaacatta cgagcaatat
     4141 gaatcgctac aagaaagaac ataccgttgc ggtagaatgg ttagctgaca aacgcgtgtg
     4201 ggctgctggt gtatactttg tacaccgcgt caactccgat atcctgttca agcgccttaa
     4261 ccagaatgtt tcacgtcacc gctcgcttga ggtaacaaag caggaagtaa ttaagaagct
     4321 gtcagggggc gaggacgata tcgctatgga ccgtcttaat atctctttat tagacccgct
     4381 gtgtaagact cgtatgacga caccgtcgcg ctgtcaagac tgtacccatt tgcaatgctt
     4441 cgacttactg tcctacctga tgatgaacga aaaaaaacca acttggcaat gccctgtgtg
     4501 tagctccaat tgtccgtacg atcgtttgat tgtcgacgac tattttttgg atatgcttgc
     4561 aaaagtagac aaaaatacta ccgaagttga actgaaagag gacgggtcct acgatgtaat
     4621 caaagaggag gctttttgta tctcagatga cgatgatgac gacgttgtgc cggccacagt
     4681 taatgggacg gcctcatgct ccagtacgaa cgggaacggg ctggccaatg aagctgctaa
     4741 aaaaaagccc gcagacgacg acattattac tctttcggat gacgatgatg aagaattgaa
     4801 tcgtggcatc atgaattcgt tgtagcgagc tccgtcgaca agcttgcggc cgcactcgag
     4861 caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg
     4921 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     4981 aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta
     5041 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     5101 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     5161 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     5221 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     5281 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     5341 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     5401 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     5461 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     5521 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgaatt aattcttaga
     5581 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
     5641 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
     5701 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
     5761 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
     5821 ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat
     5881 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
     5941 gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
     6001 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
     6061 ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag
     6121 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     6181 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     6241 ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat
     6301 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctag
     6361 agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag
     6421 cagacagttt tattgttcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6481 tcag
//
