LOCUS       sequence-497717-        5885 bp    DNA     circular SYN 16-JAN-2026
DEFINITION  Expresses yeast Isc10 fused to maltose binding protein in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5885)
  AUTHORS   Rimal A, Kamdar ZP, Tio CW, Winter E
  TITLE     Isc10, an Inhibitor That Links the Anaphase-Promoting Complex to a 
            Meiosis-Specific Mitogen-Activated Protein Kinase.
  JOURNAL   Mol Cell Biol. 2020 Jul 29;40(16). pii: MCB.00097-20. doi: 
            10.1128/MCB.00097-20. Print 2020 Jul 29.
  PUBMED    32423992
REFERENCE   2  (bases 1 to 5885)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 16, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell 
            Biol."; date: "2020-07-29"; volume: "40(16). pii: MCB.00097-20. 
            doi"; pages: " 10.1128/MCB.00097-20. Print 2020 Jul 29"
COMMENT     Sequence Label: pZK6
FEATURES             Location/Qualifiers
     source          1..5885
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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)."
     CDS             77..1177
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDAALAAAQT"
     CDS             1190..1213
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     promoter        2089..2107
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2108..2132
                     /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)."
     CDS             2241..2285
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2323..2341)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2337..2384
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2605..3264)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3179..3198
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3265..3367)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3759..3776)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3893..4438)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(3973..3992)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4663..5745)
                     /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     5707..5726
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5746..5823)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(5869..5885,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atggcaatga aaatcgaaga aggtaaactg gtaatctgga ttaacggcga
      121 taaaggctat aacggtctcg ctgaagtcgg taagaaattc gagaaagata ccggaattaa
      181 agtcaccgtt gagcatccgg ataaactgga agagaaattc ccacaggttg cggcaactgg
      241 cgatggccct gacattatct tctgggcaca cgaccgcttt ggtggctacg ctcaatctgg
      301 cctgttggct gaaatcaccc cggacaaagc gttccaggac aagctgtatc cgtttacctg
      361 ggatgccgta cgttacaacg gcaagctgat tgcttacccg atcgctgttg aagcgttatc
      421 gctgatttat aacaaagatc tgctgccgaa cccgccaaaa acctgggaag agatcccggc
      481 gctggataaa gaactgaaag cgaaaggtaa gagcgcgctg atgttcaacc tgcaagaacc
      541 gtacttcacc tggccgctga ttgctgctga cgggggttat gcgttcaagt atgaaaacgg
      601 caagtacgac attaaagacg tgggcgtgga taacgctggc gcgaaagcgg gtctgacctt
      661 cctggttgac ctgattaaaa acaaacacat gaatgcagac accgattact ccatcgcaga
      721 agctgccttt aataaaggcg aaacagcgat gaccatcaac ggcccgtggg catggtccaa
      781 catcgacacc agcaaagtga attatggtgt aacggtactg ccgaccttca agggtcaacc
      841 atccaaaccg ttcgttggcg tgctgagcgc aggtattaac gccgccagtc cgaacaaaga
      901 gctggcaaaa gagttcctcg aaaactatct gctgactgat gaaggtctgg aagcggttaa
      961 taaagacaaa ccgctgggtg ccgtagcgct gaagtcttac gaggaagagt tggcgaaaga
     1021 tccacgtatt gccgccacta tggaaaacgc ccagaaaggt gaaatcatgc cgaacatccc
     1081 gcagatgtcc gctttctggt atgccgtgcg tactgcggtg atcaacgccg ccagcggtcg
     1141 tcagactgtc gatgcagccc tggcagccgc gcagactaat gccgccgcac tggaagttct
     1201 gttccagggg cccgctagca tggacgttga cgaaagattg caccaagacg agaaccagac
     1261 ccacccgttt agccagaaaa agagcagcag cttcctgatt aaggagaaag cggctaccaa
     1321 aagcaaggat ctggaacata ttcgtctgcg tgacctgaat tttaaccacc gtaaaaagct
     1381 ggatgacaaa aagctggcga aacaaattcc ggttaaggct aacttcaaaa agccgaatga
     1441 aatcgaagca cgtgcgaatc tgaacagcaa tgaactgacc gacattaacc tggattacat
     1501 cccggacagc ccgagcattg aaaagatcag cggtccggaa gatagcatcg ttgtgacccc
     1561 gcgtaacatt atccatctgc agagcgacag cgatattatc ctggaagaat gcgaacacaa
     1621 ttacgactgt agcccgtttt accgtctgtt caactatgaa aatcgtattg aaccggatga
     1681 ctacgaagct attatcaatg caatcatcac cgatgaaatc gcaggcacct acccggtttt
     1741 tgaacgtgaa ctggaatatc aagaactgaa aagcctggtg cgtaagcgtg attacatcat
     1801 gtactatttt ctgagccgtg actatcgtgg tttctttcag ctgaaagaag aacgtaccct
     1861 gttctaccgt tatccgagca tcgcttacac cagcccgctg cgttatctgg acaacggtag
     1921 cgaaaccgaa caattcaccg gtgacgacga tgaagaactg cagagctttg atttcgaaaa
     1981 caccagcagc gtgcgtaccc tggacagcaa tatttggcgt tgagcggccg cataatgctt
     2041 aagtcgaaca gaaagtaatc gtattgtaca cggccgcata atcgaaatta atacgactca
     2101 ctatagggga attgtgagcg gataacaatt ccccatctta gtatattagt taagtataag
     2161 aaggagatat acatatggca gatctcaatt ggatatcggc cggccacgcg atcgctgacg
     2221 tcggtaccct cgagtctggt aaagaaaccg ctgctgcgaa atttgaacgc cagcacatgg
     2281 actcgtctac tagcgcagct taattaacct aggctgctgc caccgctgag caataactag
     2341 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa cctcaggcat
     2401 ttgagaagca cacggtcaca ctgcttccgg tagtcaataa accggtaaac cagcaataga
     2461 cataagcggc tatttaacga ccctgccctg aaccgacgac cgggtcgaat ttgctttcga
     2521 atttctgcca ttcatccgct tattatcact tattcaggcg tagcaaccag gcgtttaagg
     2581 gcaccaataa ctgccttaaa aaaattacgc cccgccctgc cactcatcgc agtactgttg
     2641 taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat
     2701 cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatag tgaaaacggg
     2761 ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg
     2821 attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc
     2881 accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta
     2941 ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg
     3001 aacactatcc catatcacca gctcaccgtc tttcattgcc atacggaact ccggatgagc
     3061 attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt
     3121 tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc
     3181 aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt
     3241 atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc tcgataactc
     3301 aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg
     3361 ccgatcaacg tctcattttc gccaaaagtt ggcccagggc ttcccggtat caacagggac
     3421 accaggattt atttattctg cgaagtgatc ttccgtcaca ggtatttatt cggcgcaaag
     3481 tgcgtcgggt gatgctgcca acttactgat ttagtgtatg atggtgtttt tgaggtgctc
     3541 cagtggcttc tgtttctatc agctgtccct cctgttcagc tactgacggg gtggtgcgta
     3601 acggcaaaag caccgccgga catcagcgcg agcggagtgt atactggctt actatgttgg
     3661 cactgatgag ggtgtcagtg aagtgcttca tgtggcagga gaaaaaaggc tgcaccggtg
     3721 cgtcagcaga atatgtgata caggatatat tccgcttcct cgctcactga ctcgctacgc
     3781 tcggtcgttc gactgcggcg agcggaaatg gcttacgaac ggggcggaga tttcctggaa
     3841 gatgccagga agatacttaa cagggaagtg agagggccgc ggcaaagccg tttttccata
     3901 ggctccgccc ccctgacaag catcacgaaa tctgacgctc aaatcagtgg tggcgaaacc
     3961 cgacaggact ataaagatac caggcgtttc cccctggcgg ctccctcgtg cgctctcctg
     4021 ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg tctcattcca
     4081 cgcctgacac tcagttccgg gtaggcagtt cgctccaagc tggactgtat gcacgaaccc
     4141 cccgttcagt ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggaa
     4201 agacatgcaa aagcaccact ggcagcagcc actggtaatt gatttagagg agttagtctt
     4261 gaagtcatgc gccggttaag gctaaactga aaggacaagt tttggtgact gcgctcctcc
     4321 aagccagtta cctcggttca aagagttggt agctcagaga accttcgaaa aaccgccctg
     4381 caaggcggtt ttttcgtttt cagagcaaga gattacgcgc agaccaaaac gatctcaaga
     4441 agatcatctt attaatcaga taaaatattt ctagatttca gtgcaattta tctcttcaaa
     4501 tgtagcacct gaagtcagcc ccatacgata taagttgtaa ttctcatgtt agtcatgccc
     4561 cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc
     4621 ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag
     4681 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4741 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt
     4801 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
     4861 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt
     4921 atcgtcgtat cccactaccg agatgtccgc accaacgcgc agcccggact cggtaatggc
     4981 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
     5041 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
     5101 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
     5161 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
     5221 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
     5281 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
     5341 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
     5401 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
     5461 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
     5521 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
     5581 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
     5641 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
     5701 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
     5761 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt
     5821 gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaaatta atacgactca
     5881 ctata
//
