LOCUS       sequence-458110-        8053 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Bacterial expression plasmid of MBD2∆N(MBD2b) lacking the 
            N-terminus: aa 1-152. C-terminal tagged to intein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8053)
  TITLE     MBD2 modulates heterchromatin architecture and composition
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8053)
  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            "Unpublished"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(MBD2b) pc4787
FEATURES             Location/Qualifiers
     source          1..8053
                     /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"
     primer_bind     3789..3808
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3789..3807
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3808..3832
                     /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             3847..3869
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4687..5505
                     /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             5961..5972
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             6082..6237
                     /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(6302..6320)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6316..6363
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        6423..6527
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6528..6596,6597..7388)
                     /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(6746..6765)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7430..7943)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7625..7646)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7837..7856
                     /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 ggaattgggg atcggaatta attcccggtt taaaccgggg atctcgatcc
     3781 cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga
     3841 aataattttg tttaacttta agaaggagat atacatatga tggactgccc ggccctcccc
     3901 cccggatgga agaaggagga agtgatccga aaatcagggc tcagtgctgg caagagcgat
     3961 gtctactact tcagtccaag tggtaagaag ttcagaagta aacctcagct ggcaagatac
     4021 ctgggaaatg ctgttgacct tagcagtttt gacttcagga ccggcaagat gatgcctagt
     4081 aaattacaga agaacaagca gagactccgg aatgaccccc tcaatcagaa caagggtaaa
     4141 ccagacctga acacaacatt gccaattaga caaactgcat caattttcaa gcaaccagta
     4201 accaaattca cgaaccaccc gagcaataag gtgaagtcag acccccagcg gatgaatgaa
     4261 caaccacgtc agcttttctg ggagaagagg ctacaaggac ttagcgcatc agatgtaaca
     4321 gaacaaatta taaaaaccat ggagctacct aaaggtcttc aaggagtcgg tccaggtagc
     4381 aatgacgaga cccttctgtc tgctgtggcc agtgctttac acacaagctc tgcgcccatc
     4441 acaggacaag tctctgctgc cgtggaaaag aaccctgctg tttggcttaa cacatctcaa
     4501 cccctctgca aagctttcat tgttacagat gaagacatta ggaaacagga agagcgagtc
     4561 caacaagtac gcaagaaact ggaggaggca ctgatggccg acatcctgtc ccgggctgcg
     4621 gacacggagg aagtagacat tgacatggac gatggagatg aggcggaatt cctcgagggc
     4681 tcttcctgct ttgccaaggg taccaatgtt ttaatggcgg atgggtctat tgaatgtatt
     4741 gaaaacattg aggttggtaa taaggtcatg ggtaaagatg gcagacctcg tgaggtaatt
     4801 aaattgccca gaggaagaga aactatgtac agcgtcgtgc agaaaagtca gcacagagcc
     4861 cacaaaagtg actcaagtcg tgaagtgcca gaattactca agtttacgtg taatgcgacc
     4921 catgagttgg ttgttagaac acctcgtagt gtccgccgtt tgtctcgtac cattaagggt
     4981 gtcgaatatt ttgaagttat tacttttgag atgggccaaa agaaagcccc cgacggtaga
     5041 attgttgagc ttgtcaagga agtttcaaag agctacccaa tatctgaggg gcctgagaga
     5101 gccaacgaat tagtagaatc ctatagaaag gcttcaaata aagcttattt tgagtggact
     5161 attgaggcca gagatctttc tctgttgggt tcccatgttc gtaaagctac ctaccagact
     5221 tacgctccaa ttctttatga gaatgaccac tttttcgact acatgcaaaa aagtaagttt
     5281 catctcacca ttgaaggtcc aaaagtactt gcttatttac ttggtttatg gattggtgat
     5341 ggattgtctg acagggcaac tttttcggtt gattccagag atacttcttt gatggaacgt
     5401 gttactgaat atgctgaaaa gttgaatttg tgcgccgagt ataaggacag aaaagaacca
     5461 caagttgcca aaactgttaa tttgtactct aaagttgtca gaggtaatgg tattcgcaat
     5521 aatcttaata ctgagaatcc attatgggac gctattgttg gcttaggatt cttgaaggac
     5581 ggtgtcaaaa atattccttc tttcttgtct acggacaata tcggtactcg tgaaacattt
     5641 cttgctggtc taattgattc tgatggctat gttactgatg agcatggtat taaagcaaca
     5701 ataaagacaa ttcatacttc tgtcagagat ggtttggttt cccttgctcg ttctttaggc
     5761 ttagtagtct cggttaacgc agaacctgct aaggttgaca tgaatgtcac caaacataaa
     5821 attagttatg ctatttatat gtctggtgga gatgttttgc ttaacgttct ttcgaagtgt
     5881 gccggctcta aaaaattcag gcctgctccc gccgctgctt ttgcacgtga gtgccgcgga
     5941 ttttatttcg agttacaaga attgaaggaa gacgattatt atgggattac tttatctgat
     6001 gattctgatc atcagttttt gcttggatcc caggttgtcg tccatgcatg cggtggcctg
     6061 accggtctga actcaggcct cacgacaaat cctggtgtat ccgcttggca ggtcaacaca
     6121 gcttatactg cgggacaatt ggtcacatat aacggcaaga cgtataaatg tttgcagccc
     6181 cacacctcct tggcaggatg ggaaccatcc aacgttcctg ccttgtggca gcttcaatga
     6241 ctgcaggaag gggatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     6301 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     6361 ttgctgaaag gaggaactat atccggataa ctacgtcagg tggcactttt cggggaaatg
     6421 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     6481 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     6541 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     6601 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     6661 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgtttcc
     6721 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
     6781 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     6841 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     6901 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     6961 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     7021 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     7081 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     7141 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     7201 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     7261 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     7321 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     7381 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta ccccggttga
     7441 taatcagaaa agccccaaaa acaggaagat tgtataagca aatatttaaa ttgtaaacgt
     7501 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     7561 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag cccgagatag ggttgagtgt
     7621 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     7681 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcacccaaat caagtttttt
     7741 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     7801 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     7861 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     7921 taatgcgccg ctacagggcg cgtaaaagga tctaggtgaa gatccttttt gataatctca
     7981 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     8041 tcaaaggatc ttc
//
