LOCUS       sequence-175026-        5553 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-tagged smo-1(GG), 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 5553)
  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 5553)
  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-SMO-1(GG)
FEATURES             Location/Qualifiers
     source          1..5553
                     /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"
     primer_bind     3034..3053
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3073..3150)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3356..3375
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3463..3482
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3463..3481
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3482..3506
                     /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             3521..3543
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3563..3580
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3602..3622
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3920..3937
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3990..4008)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4004..4051
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4088..4543
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4175..4194)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4385..4406
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4635..5450)
                     /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(4635..5450)
                     /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     5358..5377
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5442..5461
                     /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 gcagggttgg tggttaacgg
     2221 cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga tgtccgcacc
     2281 aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc
     2341 aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc
     2401 ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag
     2461 atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa
     2521 cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc
     2581 ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc
     2641 cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt
     2701 aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc
     2761 gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga
     2821 tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc
     2881 aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag
     2941 ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt
     3001 caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt
     3061 tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc
     3121 gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact
     3181 cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga
     3241 atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac
     3301 ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
     3361 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
     3421 gtccggcgta gaggatcgag atcgatctcg atcccgcgaa attaatacga ctcactatag
     3481 gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac tttaagaagg
     3541 agatatacat atgtcgtact accatcacca tcaccatcac gattacgata tcccaacgac
     3601 cgaaaacctg tattttcagg gcgccatggc cgatgatgca gctcaagcag gcgacaacgc
     3661 cgaatacatc aagatcaagg tcgttggaca ggacagcaac gaagtgcact tccgtgtaaa
     3721 gtatggaacc tctatggcca agctgaagaa gtcgtacgct gacagaaccg gtgtcgccgt
     3781 caacagtctt cgtttcctct tcgacggacg tcgcatcaac gacgatgaca caccgaagac
     3841 gctcgagatg gaggacgatg acgtcatcga ggtctaccaa gagcagctgg gcggatgaaa
     3901 gcttgcggcc gcactcgagc accaccacca ccaccactga gatccggctg ctaacaaagc
     3961 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     4021 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc
     4081 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4141 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4201 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4261 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4321 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4381 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4441 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4501 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     4561 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4621 tcatgaatta attcttagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata
     4681 tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca
     4741 ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca
     4801 acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca
     4861 ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact
     4921 tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta
     4981 ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta
     5041 caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca
     5101 cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg
     5161 agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat
     5221 tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg
     5281 ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca
     5341 cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg
     5401 gaatttaatc gcggcctaga gcaagacgtt tcccgttgaa tatggctcat aacacccctt
     5461 gtattactgt ttatgtaagc agacagtttt attgttcatg accaaaatcc cttaacgtga
     5521 gttttcgttc cactgagcgt cagaccccgt aga
//
