LOCUS       sequence-459929-        8647 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Bacterial expression plasmid of MBD2∆GR. The Protein is lacking the 
            glysine/arginine rich domain: aa 75-96. C-terminal tagged to 
            intein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8647)
  TITLE     MBD2 modulates heterchromatin architecture and composition
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8647)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pTYB1-MBD2∆GR pc5078
FEATURES             Location/Qualifiers
     source          1..8647
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(782..801)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     942..964
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(958..1149)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1706..2788)
                     /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     2750..2769
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2789..2866)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      3019..3105
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3202..3288
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3385..3471
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3568..3654
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3751..3837
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3972..3991
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3972..3990
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3991..4015
                     /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             4030..4052
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5281..6099
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae VMA1"
                     /product="5' region of the intein from the yeast Vma1 
                     subunit of the vacuolar ATPase (Chong et al., 1998)"
                     /label=Sce VMA intein 5' region
                     /translation="CFAKGTNVLMADGSIECIENIEVGNKVMGKDGRPREVIKLPRGRE
                     TMYSVVQKSQHRAHKSDSSREVPELLKFTCNATHELVVRTPRSVRRLSRTIKGVEYFEV
                     ITFEMGQKKAPDGRIVELVKEVSKSYPISEGPERANELVESYRKASNKAYFEWTIEARD
                     LSLLGSHVRKATYQTYAPILYENDHFFDYMQKSKFHLTIEGPKVLAYLLGLWIGDGLSD
                     RATFSVDSRDTSLMERVTEYAEKLNLCAEYKDRKEPQVAKTVNLYSKVVRG"
     CDS             6555..6566
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             6676..6831
                     /codon_start=1
                     /gene="Bacillus circulans chiA"
                     /product="chitin binding domain from chitinase A1 (Watanabe
                     et al., 1994)"
                     /label=CBD
                     /translation="TTNPGVSAWQVNTAYTAGQLVTYNGKTYKCLQPHTSLAGWEPSNV
                     PALWQLQ"
     primer_bind     complement(6896..6914)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6910..6957
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        7017..7121
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7122..7190,7191..7982)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7340..7359)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(8024..8537)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8219..8240)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8431..8450
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagctatgg tgcactctca
      781 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga
      841 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
      901 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
      961 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
     1021 gtcgtgcagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
     1081 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
     1141 tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat
     1201 gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga
     1261 acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca
     1321 gggtcaatgc cagccgaacg ccagcaagac gtagcccagc gcgtcggccg ccatgccggc
     1381 gataatggcc tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc
     1441 gagggcgtgc aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg
     1501 aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat
     1561 gataaagaag acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga
     1621 gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag
     1681 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     1741 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca
     1801 gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc
     1861 cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt
     1921 tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta
     1981 ccgagatatc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg
     2041 ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca
     2101 tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa
     2161 tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac
     2221 ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc
     2281 ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga
     2341 catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt
     2401 catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg
     2461 ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca
     2521 gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac
     2581 tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt
     2641 tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa
     2701 cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg
     2761 cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc
     2821 gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc
     2881 tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc
     2941 accgccgccg caaggaatgg tgcatgccgg catgccgccc tttcgtcttc aagaattaat
     3001 tcccaattcc ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     3061 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat
     3121 ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc
     3181 aggaattaat tccccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     3241 cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg
     3301 gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact
     3361 gccaggaatt aattccccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
     3421 tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga
     3481 gcggatttga acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa
     3541 actgccagga attaattccc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     3601 gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg
     3661 ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca
     3721 taaactgcca ggaattaatt ccccaggcat caaataaaac gaaaggctca gtcgaaagac
     3781 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
     3841 ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg
     3901 ccataaactg ccaggaattg gggatcggaa ttaattcccg gtttaaaccg gggatctcga
     3961 tcccgcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctct
     4021 agaaataatt ttgtttaact ttaagaagga gatatacata tgtcaaaggc ctacgtcgac
     4081 gccatgcgcg cgcacccggg gggaggccgc tgctgcccgg agcaggagga gggggagagc
     4141 gcggcgggcg gcagcggcgc tggcggcgac tccgccatag agcagggggg ccagggcagc
     4201 gcgctcgctc cgtccccggt gagcggcgtg cgcagggaag gcgctcgggg cggcggccgt
     4261 ggccgggggc ggtggaagca ggcggcccgg ggcggcggcg tctgtcccca gagtggcggc
     4321 agcggccttg gcggcgacgg cggcggcggc gcgggcggct gcggcgtcgg cagcggtggc
     4381 ggcgtcgccc cccggcggga tcctgtccct ttcccgtcgg ggagctcggg gccggggccc
     4441 aggggacccc gggccacgga gagcgggaag aggatggact gcccggccct cccccccgga
     4501 tggaagaagg aggaagtgat ccgaaaatca gggctcagtg ctggcaagag cgatgtctac
     4561 tacttcagtc caagtggtaa gaagttcaga agtaaacctc agctggcaag atacctggga
     4621 aatgctgttg accttagcag ttttgacttc aggaccggca agatgatgcc tagtaaatta
     4681 cagaagaaca agcagagact ccggaatgac cccctcaatc agaacaaggg taaaccagac
     4741 ctgaacacaa cattgccaat tagacaaact gcatcaattt tcaagcaacc agtaaccaaa
     4801 ttcacgaacc acccgagcaa taaggtgaag tcagaccccc agcggatgaa tgaacaacca
     4861 cgtcagcttt tctgggagaa gaggctacaa ggacttagcg catcagatgt aacagaacaa
     4921 attataaaaa ccatggagct acctaaaggt cttcaaggag tcggtccagg tagcaatgac
     4981 gagacccttc tgtctgctgt ggccagtgct ttacacacaa gctctgcgcc catcacagga
     5041 caagtctctg ctgccgtgga aaagaaccct gctgtttggc ttaacacatc tcaacccctc
     5101 tgcaaagctt tcattgttac agatgaagac attaggaaac aggaagagcg agtccaacaa
     5161 gtacgcaaga aactggagga ggcactgatg gccgacatcc tgtcccgggc tgcggacacg
     5221 gaggaagtag acattgacat ggacgatgga gatgaggcgg aattcctcga gggctcttcc
     5281 tgctttgcca agggtaccaa tgttttaatg gcggatgggt ctattgaatg tattgaaaac
     5341 attgaggttg gtaataaggt catgggtaaa gatggcagac ctcgtgaggt aattaaattg
     5401 cccagaggaa gagaaactat gtacagcgtc gtgcagaaaa gtcagcacag agcccacaaa
     5461 agtgactcaa gtcgtgaagt gccagaatta ctcaagttta cgtgtaatgc gacccatgag
     5521 ttggttgtta gaacacctcg tagtgtccgc cgtttgtctc gtaccattaa gggtgtcgaa
     5581 tattttgaag ttattacttt tgagatgggc caaaagaaag cccccgacgg tagaattgtt
     5641 gagcttgtca aggaagtttc aaagagctac ccaatatctg aggggcctga gagagccaac
     5701 gaattagtag aatcctatag aaaggcttca aataaagctt attttgagtg gactattgag
     5761 gccagagatc tttctctgtt gggttcccat gttcgtaaag ctacctacca gacttacgct
     5821 ccaattcttt atgagaatga ccactttttc gactacatgc aaaaaagtaa gtttcatctc
     5881 accattgaag gtccaaaagt acttgcttat ttacttggtt tatggattgg tgatggattg
     5941 tctgacaggg caactttttc ggttgattcc agagatactt ctttgatgga acgtgttact
     6001 gaatatgctg aaaagttgaa tttgtgcgcc gagtataagg acagaaaaga accacaagtt
     6061 gccaaaactg ttaatttgta ctctaaagtt gtcagaggta atggtattcg caataatctt
     6121 aatactgaga atccattatg ggacgctatt gttggcttag gattcttgaa ggacggtgtc
     6181 aaaaatattc cttctttctt gtctacggac aatatcggta ctcgtgaaac atttcttgct
     6241 ggtctaattg attctgatgg ctatgttact gatgagcatg gtattaaagc aacaataaag
     6301 acaattcata cttctgtcag agatggtttg gtttcccttg ctcgttcttt aggcttagta
     6361 gtctcggtta acgcagaacc tgctaaggtt gacatgaatg tcaccaaaca taaaattagt
     6421 tatgctattt atatgtctgg tggagatgtt ttgcttaacg ttctttcgaa gtgtgccggc
     6481 tctaaaaaat tcaggcctgc tcccgccgct gcttttgcac gtgagtgccg cggattttat
     6541 ttcgagttac aagaattgaa ggaagacgat tattatggga ttactttatc tgatgattct
     6601 gatcatcagt ttttgcttgg atcccaggtt gtcgtccatg catgcggtgg cctgaccggt
     6661 ctgaactcag gcctcacgac aaatcctggt gtatccgctt ggcaggtcaa cacagcttat
     6721 actgcgggac aattggtcac atataacggc aagacgtata aatgtttgca gccccacacc
     6781 tccttggcag gatgggaacc atccaacgtt cctgccttgt ggcagcttca atgactgcag
     6841 gaaggggatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     6901 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
     6961 aaaggaggaa ctatatccgg ataactacgt caggtggcac ttttcgggga aatgtgcgcg
     7021 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     7081 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     7141 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     7201 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     7261 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt ttcccaatga
     7321 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     7381 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     7441 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     7501 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     7561 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     7621 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     7681 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     7741 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     7801 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     7861 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     7921 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     7981 aactgtcaga ccaagtttac tcatatatac tttagattga tttaccccgg ttgataatca
     8041 gaaaagcccc aaaaacagga agattgtata agcaaatatt taaattgtaa acgttaatat
     8101 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     8161 aatcggcaaa atcccttata aatcaaaaga atagcccgag atagggttga gtgttgttcc
     8221 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     8281 cgtctatcag ggcgatggcc cactacgtga accatcaccc aaatcaagtt ttttggggtc
     8341 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     8401 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     8461 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     8521 gccgctacag ggcgcgtaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     8581 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     8641 gatcttc
//
