LOCUS       Exported                6062 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  E.Coli produced TET1-848-952 purified by FPLC nickel column.
ACCESSION   .
VERSION     .
KEYWORDS    GB1-His-TET1-848-952(pL91)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6062)
  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 6062)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6062
                     /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             668..685
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      851..937
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1029..1056
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1110..1128)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1124..1171
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1208..1663
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1295..1314)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1505..1526
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1778..2638)
                     /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     2401..2420
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2639..2731)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2812..3400
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3301..3320
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3554..3571
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3586..3725
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3651..3670)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3811..3833
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3827..4018)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4590..5672)
                     /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     5634..5653
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5673..5750)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5956..5975
                     /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 gctagtatac acaatgaagg tgatcaacca aaaactcctg agaatatacc
      361 aagtaaagaa ccaaaagatg gatctcccgt tcaaccaagt ctcttatcgt taatgaaaga
      421 taggagatta acattggagc aagtggtagc catagaggcc ctgactcaac tctcagaagc
      481 cccatcagag aattcctccc catcaaagtc agagaaggat gaggaatcag agcagagaac
      541 agccagtttg cttaatagct gcaaagctat cctctacact gtaagaaaag acctccaaga
      601 cccaaactta cagggagagc caccatgaaa ttcgagctcc gtcgacaagc ttgcggccgc
      661 actcgagcac caccaccacc accactaatg ttaattaagt tgggcgttcc taggctgata
      721 aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca
      781 gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag agtagggaac
      841 tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
      901 ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt
      961 tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca
     1021 aattaagcag aaggccatcc tgacggatgg cctttttgcg tttctacaaa ctcttttgtt
     1081 tatttttcta aatacattca aatatgtatc cgctgagcaa taactagcat aaccccttgg
     1141 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc
     1201 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     1261 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     1321 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     1381 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     1441 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     1501 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     1561 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     1621 aaatttaacg cgaattttaa caaaatatta acgtttacaa tttctggcgg cacgatggca
     1681 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     1741 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     1801 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     1861 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     1921 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     1981 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     2041 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     2101 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     2161 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     2221 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     2281 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     2341 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     2401 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     2461 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     2521 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     2581 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     2641 tcttcctttt tcaatcatga ttgaagcatt tatcagggtt attgtctcat gagcggatac
     2701 atatttgaat gtatttagaa aaataaacaa ataggtcatg accaaaatcc cttaacgtga
     2761 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     2821 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     2881 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     2941 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3001 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3061 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3121 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3181 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3241 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3301 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     3361 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     3421 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     3481 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     3541 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     3601 tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc
     3661 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     3721 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3781 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3841 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     3901 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     3961 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg
     4021 atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga
     4081 ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg
     4141 gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc
     4201 agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc
     4261 agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac
     4321 ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc
     4381 ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc
     4441 ctagccgggt cctcaacgac aggagcacga tcatgctagt catgccccgc gcccaccgga
     4501 aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag
     4561 tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     4621 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4681 gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc
     4741 tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc
     4801 tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc
     4861 actaccgaga tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc
     4921 agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt
     4981 tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc
     5041 tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca
     5101 gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc
     5161 acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca
     5221 gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc
     5281 tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc
     5341 accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca
     5401 cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc
     5461 agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg
     5521 cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca
     5581 gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac
     5641 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc
     5701 gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg
     5761 acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt
     5821 gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc
     5881 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc
     5941 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc
     6001 ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atcgatctcg atcccgcgaa
     6061 at
//
