LOCUS       sequence-193502-        6590 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression for structure determination; may not be full 
            ORF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6590)
  TITLE     SGC Toronto Q4 2016, Q1 Q2 2017
REFERENCE   2  (bases 1 to 6590)
  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"
COMMENT     Sequence Label: 5WCI (MYST1)
FEATURES             Location/Qualifiers
     source          1..6590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             91..108
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             121..141
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1455..1472
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1525..1543)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1539..1586
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1623..2078
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1710..1729)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1920..1941
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2170..2985)
                     /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(2170..2985)
                     /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     2893..2912
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2977..2996
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3107..3695
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3596..3615
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3849..3866
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3881..4020
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3946..3965)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4106..4128
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4122..4313)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5122..6204)
                     /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     6166..6185
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6205..6282)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6488..6507
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg catcatcatc atcatcacag cagcggcaga
      121 gaaaacttgt atttccaggg ccgcgtctct ccgccgaccc cggcgcgcgg cgagccggaa
      181 gtcacggtgg agatcggaga aacgtacctg tgccggcgac cggatagcac ctggcattct
      241 gctgaagtga tccagtctcg agtgaacgac caggagggcc gagaggaatt ctatgtacac
      301 tacgtgggct ttaaccggcg gctggacgag tgggtagaca agaaccggct ggcgctgacc
      361 aagacagtga aggatgctgt acagaagaac tcagagaagt acctgagcga gctcgcagag
      421 cagcctgagc gcaagatcac tcgcaaccaa aagcgcaagc atgatgagat caaccatgtg
      481 cagaagactt atgcagagat ggaccccacc acagcagcct tggagaagga gcatgaggcg
      541 atcaccaagg tgaagtatgt ggacaagatc cacatcggga actacgaaat tgatgcctgg
      601 tatttctcac cattccccga agactatggg aaacagccca agctctggct ctgcgagtac
      661 tgcctcaagt acatgaaata tgagaagagc taccgcttcc acttgggtca gtgccagtgg
      721 cggcagcccc ccgggaaaga gatctaccgc aagagcaaca tctccgtgta cgaagttgat
      781 ggcaaagacc ataagattta ctgtcagaac ctgtgtctgc tggccaagct tttcctggac
      841 cataagacac tgtactttga cgtggagccg ttcgtctttt acatcctgac tgaggtggac
      901 cggcaggggg cccacattgt tggctacttc tccaaggaga aggagtcccc ggatggaaac
      961 aatgtggcct gcatcctgac cttgcccccc taccaacgcc gcggctacgg gaagttcctc
     1021 atcgctttca gttatgagct ctccaagctg gagagcacag tcggctcccc ggagaagccg
     1081 ctgtctgacc tgggcaagct cagctaccgc agctactggt cctgggtgct gctggagatc
     1141 ctgcgggact tccggggcac actgtccatc aaggacctca gccagatgac cagtatcacc
     1201 caaaatgaca tcatcagtac cctgcaatcc ctcaatatgg tcaagtactg gaagggccag
     1261 cacgtgatct gtgtcacacc caagctggtg gaggagcacc tcaaaagtgc ccagtataag
     1321 aaaccaccca tcacaggtgg gtggggggct gctgtgtgtc gggggcggtg ggggagtgtc
     1381 agtatgtgga ctggtaggag tcaaggcctc cttattgctg tcacatgatg acgaagcttg
     1441 cggccgcact cgagcaccac caccaccacc actgagatcc ggctgctaac aaagcccgaa
     1501 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     1561 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga ttggcgaatg
     1621 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     1681 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     1741 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     1801 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     1861 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     1921 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     1981 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     2041 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     2101 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2161 aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg
     2221 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
     2281 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
     2341 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
     2401 agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc
     2461 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
     2521 tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac
     2581 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga
     2641 atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa
     2701 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
     2761 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     2821 tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga
     2881 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     2941 taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt
     3001 actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa cgtgagtttt
     3061 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3121 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3181 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3241 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3301 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3361 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3421 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3481 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3541 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3601 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3661 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3721 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     3781 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     3841 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     3901 ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga
     3961 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
     4021 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     4081 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     4141 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca
     4201 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc
     4261 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc
     4321 tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg
     4381 ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa
     4441 caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc
     4501 ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc
     4561 cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa
     4621 ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac
     4681 gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc
     4741 cgggtcctca acgacaggag cacgatcatg cgcacccgtg gggccgccat gccggcgata
     4801 atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg
     4861 gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag
     4921 cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag ttgcatgata
     4981 aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg
     5041 actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa
     5101 cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5161 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt
     5221 ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg
     5281 agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat
     5341 ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga
     5401 gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat
     5461 ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt
     5521 ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg
     5581 attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa
     5641 tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag
     5701 tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc
     5761 aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc
     5821 cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc
     5881 tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg
     5941 atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga
     6001 ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg
     6061 aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg
     6121 gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac
     6181 atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta
     6241 tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct cgacgctctc
     6301 ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg ttgagcaccg
     6361 ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc
     6421 ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt
     6481 ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc
     6541 cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat
//
