LOCUS       Exported                5849 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  E.Coli produced TET1-SID mutants (L900A) purified by FPLC nickel 
            column.
ACCESSION   .
VERSION     .
KEYWORDS    GB1-His-TET1-878-911-L900A(pL91)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5849)
  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 5849)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5849
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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             455..472
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      638..724
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      816..843
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(897..915)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      911..958
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      995..1450
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1082..1101)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1292..1313
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1565..2425)
                     /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     2188..2207
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2426..2518)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2599..3187
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3088..3107
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3341..3358
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3373..3512
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3438..3457)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3598..3620
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3614..3805)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4377..5459)
                     /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     5421..5440
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5460..5537)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5743..5762
                     /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 ctcttatcgt taatgaaaga taggagatta acattggagc aagtggtagc
      361 catagaggcc ctgactcaag cctcagaagc cccatcagag aattcctccc catgaaattc
      421 gagctccgtc gacaagcttg cggccgcact cgagcaccac caccaccacc actaatgtta
      481 attaagttgg gcgttcctag gctgataaaa cagaatttgc ctggcggcag tagcgcggtg
      541 gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg
      601 gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc
      661 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
      721 aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg
      781 acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga cggatggcct
      841 ttttgcgttt ctacaaactc ttttgtttat ttttctaaat acattcaaat atgtatccgc
      901 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
      961 gaaaggagga actatatccg gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     1021 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     1081 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     1141 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     1201 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     1261 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     1321 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     1381 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     1441 tttacaattt ctggcggcac gatggcatga gattatcaaa aaggatcttc acctagatcc
     1501 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     1561 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     1621 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1681 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     1741 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     1801 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     1861 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     1921 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     1981 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     2041 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     2101 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     2161 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     2221 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     2281 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     2341 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     2401 cgacacggaa atgttgaata ctcatactct tcctttttca atcatgattg aagcatttat
     2461 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     2521 ggtcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     2581 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2641 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2701 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     2761 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2821 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2881 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2941 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3001 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3061 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3121 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3181 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3241 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     3301 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     3361 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     3421 caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
     3481 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
     3541 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     3601 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     3661 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     3721 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     3781 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg
     3841 gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac
     3901 atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc
     3961 agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac
     4021 agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact
     4081 tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg
     4141 tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct
     4201 gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca
     4261 tgctagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca
     4321 tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac
     4381 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4441 cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg
     4501 ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg
     4561 ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat
     4621 gagctgtctt cggtatcgtc gtatcccact accgagatgt ccgcaccaac gcgcagcccg
     4681 gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca
     4741 gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc
     4801 cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag
     4861 ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc
     4921 tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa
     4981 ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg
     5041 caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca
     5101 ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt
     5161 tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg
     5221 acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac
     5281 tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc
     5341 gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa
     5401 acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca
     5461 ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg
     5521 cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa
     5581 gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa
     5641 ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca
     5701 agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata
     5761 ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag
     5821 gatcgagatc gatctcgatc ccgcgaaat
//
