LOCUS       sequence-193501-        5624 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 5624)
  TITLE     SGC Toronto Q4 2016, Q1 Q2 2017
REFERENCE   2  (bases 1 to 5624)
  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: 5WBN (HDAC6)
FEATURES             Location/Qualifiers
     source          1..5624
                     /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             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             127..144
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             489..506
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(559..577)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      573..620
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      657..1112
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(744..763)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     954..975
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1204..2019)
                     /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(1204..2019)
                     /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     1927..1946
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2011..2030
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2141..2729
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2630..2649
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2883..2900
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2915..3054
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2980..2999)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3140..3162
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3156..3347)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4156..5238)
                     /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     5200..5219
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5239..5316)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5522..5541
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      121 agcggcctgg ttccgcgtgg tagtccactg ccctggtgtc cccatttggt ggcagtatgc
      181 cccatacctg cagcaggcct agacgtgacc caaccttgtg gggactgtgg aacaatccaa
      241 gagaattggg tgtgtctctc ttgctatcag gtctactgtg gtcgttacat caatggccac
      301 atgctccaac accatggaaa ttctggacac ccgctggtcc tcagctacat cgacctgtca
      361 gcctggtgtt actactgtca ggcctatgtc caccaccagg ctctcctaga tgtgaagaac
      421 atcgcccacc agaacaagtt tggggaggat atgccccact gatgacgaag cttgcggccg
      481 cactcgagca ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag
      541 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
      601 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc
      661 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
      721 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
      781 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
      841 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
      901 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
      961 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     1021 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     1081 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
     1141 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgaattaa
     1201 ttcttagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
     1261 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
     1321 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
     1381 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
     1441 gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg
     1501 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
     1561 ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
     1621 cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg
     1681 atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc
     1741 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
     1801 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
     1861 aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc
     1921 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
     1981 cggcctagag caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt
     2041 tatgtaagca gacagtttta ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc
     2101 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     2161 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     2221 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     2281 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2341 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2401 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2461 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2521 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2581 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2641 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     2701 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     2761 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     2821 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     2881 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     2941 atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc
     3001 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     3061 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     3121 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     3181 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
     3241 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     3301 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     3361 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
     3421 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
     3481 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     3541 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     3601 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     3661 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
     3721 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
     3781 ctcaacgaca ggagcacgat catgcgcacc cgtggggccg ccatgccggc gataatggcc
     3841 tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc
     3901 aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc
     3961 tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag
     4021 acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg
     4081 ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca
     4141 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4201 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt
     4261 tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag
     4321 ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt
     4381 taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc
     4441 cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc
     4501 gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg
     4561 aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg
     4621 agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc
     4681 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
     4741 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
     4801 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
     4861 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
     4921 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
     4981 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
     5041 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
     5101 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
     5161 ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta
     5221 taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc
     5281 cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat
     5341 gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg
     5401 caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
     5461 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
     5521 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     5581 cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg aaat
//
