LOCUS       sequence-460687-        7906 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Bacterial expression plasmid of MBD2-N (N-terminus of MBD2: aa 
            1-152). C-terminal tagged to intein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7906)
  TITLE     MBD2 modulates heterchromatin architecture and composition
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7906)
  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            "Unpublished"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-N pc4789
FEATURES             Location/Qualifiers
     source          1..7906
                     /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             4540..5358
                     /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             5814..5825
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             5935..6090
                     /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(6155..6173)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6169..6216
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        6276..6380
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6381..6449,6450..7241)
                     /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(6599..6618)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7283..7796)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7478..7499)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7690..7709
                     /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 tgatgcgcgc gcacccgggg
     4081 ggaggccgct gctgcccgga gcaggaggag ggggagagcg cggcgggcgg cagcggcgct
     4141 ggcggcgact ccgccataga gcaggggggc cagggcagcg cgctcgctcc gtccccggtg
     4201 agcggcgtgc gcagggaagg cgctcggggc ggcggccgtg gccgggggcg gtggaagcag
     4261 gcggcccggg gcggcggcgt ctgtggccgt ggccgtggcc gtggccgggg tcggggccgt
     4321 ggccggggcc ggggccgggg ccgcggccgt ccccagagtg gcggcagcgg ccttggcggc
     4381 gacggcggcg gcggcgcggg cggctgcggc gtcggcagcg gtggcggcgt cgccccccgg
     4441 cgggatcctg tccctttccc gtcggggagc tcggggccgg ggcccagggg accccgggcc
     4501 acggagagcg ggaagaggga attcctcgag ggctcttcct gctttgccaa gggtaccaat
     4561 gttttaatgg cggatgggtc tattgaatgt attgaaaaca ttgaggttgg taataaggtc
     4621 atgggtaaag atggcagacc tcgtgaggta attaaattgc ccagaggaag agaaactatg
     4681 tacagcgtcg tgcagaaaag tcagcacaga gcccacaaaa gtgactcaag tcgtgaagtg
     4741 ccagaattac tcaagtttac gtgtaatgcg acccatgagt tggttgttag aacacctcgt
     4801 agtgtccgcc gtttgtctcg taccattaag ggtgtcgaat attttgaagt tattactttt
     4861 gagatgggcc aaaagaaagc ccccgacggt agaattgttg agcttgtcaa ggaagtttca
     4921 aagagctacc caatatctga ggggcctgag agagccaacg aattagtaga atcctataga
     4981 aaggcttcaa ataaagctta ttttgagtgg actattgagg ccagagatct ttctctgttg
     5041 ggttcccatg ttcgtaaagc tacctaccag acttacgctc caattcttta tgagaatgac
     5101 cactttttcg actacatgca aaaaagtaag tttcatctca ccattgaagg tccaaaagta
     5161 cttgcttatt tacttggttt atggattggt gatggattgt ctgacagggc aactttttcg
     5221 gttgattcca gagatacttc tttgatggaa cgtgttactg aatatgctga aaagttgaat
     5281 ttgtgcgccg agtataagga cagaaaagaa ccacaagttg ccaaaactgt taatttgtac
     5341 tctaaagttg tcagaggtaa tggtattcgc aataatctta atactgagaa tccattatgg
     5401 gacgctattg ttggcttagg attcttgaag gacggtgtca aaaatattcc ttctttcttg
     5461 tctacggaca atatcggtac tcgtgaaaca tttcttgctg gtctaattga ttctgatggc
     5521 tatgttactg atgagcatgg tattaaagca acaataaaga caattcatac ttctgtcaga
     5581 gatggtttgg tttcccttgc tcgttcttta ggcttagtag tctcggttaa cgcagaacct
     5641 gctaaggttg acatgaatgt caccaaacat aaaattagtt atgctattta tatgtctggt
     5701 ggagatgttt tgcttaacgt tctttcgaag tgtgccggct ctaaaaaatt caggcctgct
     5761 cccgccgctg cttttgcacg tgagtgccgc ggattttatt tcgagttaca agaattgaag
     5821 gaagacgatt attatgggat tactttatct gatgattctg atcatcagtt tttgcttgga
     5881 tcccaggttg tcgtccatgc atgcggtggc ctgaccggtc tgaactcagg cctcacgaca
     5941 aatcctggtg tatccgcttg gcaggtcaac acagcttata ctgcgggaca attggtcaca
     6001 tataacggca agacgtataa atgtttgcag ccccacacct ccttggcagg atgggaacca
     6061 tccaacgttc ctgccttgtg gcagcttcaa tgactgcagg aaggggatcc ggctgctaac
     6121 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     6181 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
     6241 taactacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     6301 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     6361 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     6421 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     6481 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     6541 tccttgagag ttttcgcccc gaagaacgtt tcccaatgat gagcactttt aaagttctgc
     6601 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
     6661 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     6721 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     6781 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     6841 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     6901 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     6961 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     7021 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     7081 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     7141 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     7201 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     7261 catatatact ttagattgat ttaccccggt tgataatcag aaaagcccca aaaacaggaa
     7321 gattgtataa gcaaatattt aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa
     7381 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     7441 atcaaaagaa tagcccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     7501 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     7561 actacgtgaa ccatcaccca aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     7621 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     7681 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     7741 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtaaaa
     7801 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     7861 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttc
//
