LOCUS       sequence-245109-        5930 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E.Coli produced SAP25 SID purified by FPLC nickel column.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5930)
  AUTHORS   Chandru A, Bate N, Vuister GW, Cowley SM
  TITLE     Sin3A recruits Tet1 to the PAH1 domain via a highly conserved 
            Sin3-Interaction Domain.
  JOURNAL   Sci Rep. 2018 Oct 2;8(1):14689. doi: 10.1038/s41598-018-32942-w.
  PUBMED    30279502
REFERENCE   2  (bases 1 to 5930)
  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: "Sci Rep."; 
            date: "2018-10-2"; volume: "8(1):14689. doi"; pages: " 
            10.1038/s41598-018-32942-w"
COMMENT     Sequence Label: GB1-His-SAP25-126-186(pL91)
FEATURES             Location/Qualifiers
     source          1..5930
                     /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             95..256
                     /codon_start=1
                     /product="B1 domain of Streptococcal protein G   "
                     /label=GB1
                     /note="effective as a solubilizing fusion partner (Cheng 
                     and Patel, 2004)"
                     /translation="YKLILNGKTLKGETTTEAVDAATAEKVFKQYANDNGVDGEWTYDD
                     ATKTFTVTE"
     CDS             263..280
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             287..307
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             536..553
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      719..805
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      897..924
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(978..996)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      992..1039
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1076..1531
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1163..1182)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1373..1394
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(1646..2437,2438..2506))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2269..2288
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2507..2599)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2680..3268
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3169..3188
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3422..3439
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3454..3593
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3519..3538)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3679..3701
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3695..3886)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4458..5540)
                     /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     5502..5521
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5541..5618)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5824..5843
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gcagtacaaa ctgatcctga acggtaaaac
      121 cctgaaaggt gaaaccacca ccgaagctgt tgacgctgct accgcggaaa aagttttcaa
      181 acagtacgct aacgacaacg gtgttgacgg tgaatggacc tacgacgacg ctaccaaaac
      241 cttcaccgtt accgaaggat ctcatcatca tcatcatcat ggatccgaaa acctgtattt
      301 tcagggcgcc tcctccacct ggctcagcga ggcagagatg atcgcccttg ctggtctgct
      361 gcagatgagc cagggagagc agacacccaa ctgcgtggca agctctctgc cttccaccag
      421 ctgcccagac cctgtctctg tctctgaaga cccaggtccc agtggtgacc agagctgttc
      481 tgggactgac acatgaaatt cgagctccgt cgacaagctt gcggccgcac tcgagcacca
      541 ccaccaccac cactaatgtt aattaagttg ggcgttccta ggctgataaa acagaatttg
      601 cctggcggca gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc
      661 cgtagcgccg atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca
      721 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
      781 gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg
      841 gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa
      901 ggccatcctg acggatggcc tttttgcgtt tctacaaact cttttgttta tttttctaaa
      961 tacattcaaa tatgtatccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     1021 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg
     1081 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     1141 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     1201 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     1261 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     1321 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     1381 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     1441 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     1501 aattttaaca aaatattaac gtttacaatt tctggcggca cgatggcatg agattatcaa
     1561 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     1621 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     1681 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     1741 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     1801 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     1861 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     1921 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     1981 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     2041 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     2101 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     2161 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     2221 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     2281 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     2341 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     2401 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     2461 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     2521 aatcatgatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     2581 atttagaaaa ataaacaaat aggtcatgac caaaatccct taacgtgagt tttcgttcca
     2641 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     2701 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     2761 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     2821 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     2881 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     2941 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3001 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3061 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3121 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3181 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3241 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3301 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     3361 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     3421 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     3481 tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca
     3541 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
     3601 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     3661 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     3721 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
     3781 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
     3841 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt
     3901 aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga
     3961 tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg
     4021 cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta
     4081 atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca
     4141 taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga
     4201 ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct
     4261 cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc
     4321 tcaacgacag gagcacgatc atgctagtca tgccccgcgc ccaccggaag gagctgactg
     4381 ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta
     4441 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4501 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt
     4561 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
     4621 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
     4681 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg
     4741 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga
     4801 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt
     4861 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg
     4921 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
     4981 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
     5041 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
     5101 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
     5161 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
     5221 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
     5281 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
     5341 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
     5401 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
     5461 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
     5521 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
     5581 gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt
     5641 atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc
     5701 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
     5761 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
     5821 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
     5881 cacgatgcgt ccggcgtaga ggatcgagat cgatctcgat cccgcgaaat
//
