LOCUS       sequence-459933-        7705 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Bacterial expression plasmid of MBD2-C∆CC. C-terminus of MBD2 is 
            lacking the coiled coil domain: aa 363-396. C-terminal tagged to 
            intein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7705)
  TITLE     MBD2 modulates heterchromatin architecture and composition
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7705)
  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            "Unpublished"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-C∆CC pc5084
FEATURES             Location/Qualifiers
     source          1..7705
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..69,70..861)
                     /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(219..238)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(903..1416)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1098..1119)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1310..1329
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      1527..2115
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2016..2035
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2269..2286
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2308..2327)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2468..2490
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2484..2675)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3232..4314)
                     /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     4276..4295
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4315..4392)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      4545..4631
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4728..4814
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4911..4997
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5094..5180
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5315..5334
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5315..5333
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5334..5358
                     /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             5373..5395
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5865..6683
                     /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             7139..7150
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             7260..7415
                     /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(7480..7498)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7494..7541
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        7601..7705
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
       61 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
      121 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
      181 gaagaacgtt tcccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
      241 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
      301 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
      361 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
      421 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
      481 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
      541 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
      601 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
      661 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
      721 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
      781 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
      841 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
      901 ttaccccggt tgataatcag aaaagcccca aaaacaggaa gattgtataa gcaaatattt
      961 aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     1021 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagcccgaga
     1081 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     1141 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccca
     1201 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     1261 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     1321 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     1381 cacccgccgc gcttaatgcg ccgctacagg gcgcgtaaaa ggatctaggt gaagatcctt
     1441 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     1501 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     1561 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     1621 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     1681 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     1741 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     1801 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     1861 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     1921 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     1981 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     2041 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     2101 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     2161 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     2221 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     2281 agcgaggaag ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     2341 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     2401 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     2461 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     2521 cagctgcggt aaagctcatc agcgtggtcg tgcagcgatt cacagatgtc tgcctgttca
     2581 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     2641 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     2701 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
     2761 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
     2821 cggcgggacc agagaaaaat cactcagggt caatgccagc cgaacgccag caagacgtag
     2881 cccagcgcgt cggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg
     2941 gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac
     3001 aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct
     3061 gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata
     3121 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
     3181 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     3241 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3301 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     3361 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     3421 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     3481 gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc
     3541 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     3601 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     3661 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     3721 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     3781 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     3841 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     3901 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     3961 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     4021 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     4081 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     4141 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     4201 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     4261 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     4321 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca
     4381 ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc
     4441 ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgccggcatg
     4501 ccgccctttc gtcttcaaga attaattccc aattccccag gcatcaaata aaacgaaagg
     4561 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
     4621 gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc
     4681 gggcaggacg cccgccataa actgccagga attaattccc caggcatcaa ataaaacgaa
     4741 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     4801 tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt
     4861 ggcgggcagg acgcccgcca taaactgcca ggaattaatt ccccaggcat caaataaaac
     4921 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     4981 tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag
     5041 ggtggcgggc aggacgcccg ccataaactg ccaggaatta attccccagg catcaaataa
     5101 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     5161 ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg
     5221 gagggtggcg ggcaggacgc ccgccataaa ctgccaggaa ttggggatcg gaattaattc
     5281 ccggtttaaa ccggggatct cgatcccgcg aaattaatac gactcactat aggggaattg
     5341 tgagcggata acaattcccc tctagaaata attttgttta actttaagaa ggagatatac
     5401 atatgatgaa caagggtaaa ccagacctga acacaacatt gccaattaga caaactgcat
     5461 caattttcaa gcaaccagta accaaattca cgaaccaccc gagcaataag gtgaagtcag
     5521 acccccagcg gatgaatgaa caaccacgtc agcttttctg ggagaagagg ctacaaggac
     5581 ttagcgcatc agatgtaaca gaacaaatta taaaaaccat ggagctacct aaaggtcttc
     5641 aaggagtcgg tccaggtagc aatgacgaga cccttctgtc tgctgtggcc agtgctttac
     5701 acacaagctc tgcgcccatc acaggacaag tctctgctgc cgtggaaaag aaccctgctg
     5761 tttggcttaa cacatctcaa cccctctgcc gggctgcgga cacggaggaa gtagacattg
     5821 acatggacga tggagatgag gcggaattcc tcgagggctc ttcctgcttt gccaagggta
     5881 ccaatgtttt aatggcggat gggtctattg aatgtattga aaacattgag gttggtaata
     5941 aggtcatggg taaagatggc agacctcgtg aggtaattaa attgcccaga ggaagagaaa
     6001 ctatgtacag cgtcgtgcag aaaagtcagc acagagccca caaaagtgac tcaagtcgtg
     6061 aagtgccaga attactcaag tttacgtgta atgcgaccca tgagttggtt gttagaacac
     6121 ctcgtagtgt ccgccgtttg tctcgtacca ttaagggtgt cgaatatttt gaagttatta
     6181 cttttgagat gggccaaaag aaagcccccg acggtagaat tgttgagctt gtcaaggaag
     6241 tttcaaagag ctacccaata tctgaggggc ctgagagagc caacgaatta gtagaatcct
     6301 atagaaaggc ttcaaataaa gcttattttg agtggactat tgaggccaga gatctttctc
     6361 tgttgggttc ccatgttcgt aaagctacct accagactta cgctccaatt ctttatgaga
     6421 atgaccactt tttcgactac atgcaaaaaa gtaagtttca tctcaccatt gaaggtccaa
     6481 aagtacttgc ttatttactt ggtttatgga ttggtgatgg attgtctgac agggcaactt
     6541 tttcggttga ttccagagat acttctttga tggaacgtgt tactgaatat gctgaaaagt
     6601 tgaatttgtg cgccgagtat aaggacagaa aagaaccaca agttgccaaa actgttaatt
     6661 tgtactctaa agttgtcaga ggtaatggta ttcgcaataa tcttaatact gagaatccat
     6721 tatgggacgc tattgttggc ttaggattct tgaaggacgg tgtcaaaaat attccttctt
     6781 tcttgtctac ggacaatatc ggtactcgtg aaacatttct tgctggtcta attgattctg
     6841 atggctatgt tactgatgag catggtatta aagcaacaat aaagacaatt catacttctg
     6901 tcagagatgg tttggtttcc cttgctcgtt ctttaggctt agtagtctcg gttaacgcag
     6961 aacctgctaa ggttgacatg aatgtcacca aacataaaat tagttatgct atttatatgt
     7021 ctggtggaga tgttttgctt aacgttcttt cgaagtgtgc cggctctaaa aaattcaggc
     7081 ctgctcccgc cgctgctttt gcacgtgagt gccgcggatt ttatttcgag ttacaagaat
     7141 tgaaggaaga cgattattat gggattactt tatctgatga ttctgatcat cagtttttgc
     7201 ttggatccca ggttgtcgtc catgcatgcg gtggcctgac cggtctgaac tcaggcctca
     7261 cgacaaatcc tggtgtatcc gcttggcagg tcaacacagc ttatactgcg ggacaattgg
     7321 tcacatataa cggcaagacg tataaatgtt tgcagcccca cacctccttg gcaggatggg
     7381 aaccatccaa cgttcctgcc ttgtggcagc ttcaatgact gcaggaaggg gatccggctg
     7441 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
     7501 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat
     7561 ccggataact acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     7621 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     7681 tcaataatat tgaaaaagga agagt
//
