LOCUS       sequence-459922-        8305 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Bacterial expression vector of MBD3 C-terminal tagged to the 
            intein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8305)
  TITLE     MBD2 modulates heterchromatin architecture and composition
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8305)
  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            "Unpublished"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-MBD3 pc4784
FEATURES             Location/Qualifiers
     source          1..8305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(439..952)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(634..655)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     846..865
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      1063..1651
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1552..1571
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1805..1822
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1844..1863)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2004..2026
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2020..2211)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2768..3850)
                     /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     3812..3831
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3851..3928)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      4081..4167
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4264..4350
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4447..4533
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4630..4716
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4813..4899
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5034..5053
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5034..5052
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5053..5077
                     /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             5092..5114
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             6001..6819
                     /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             7275..7286
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             7396..7551
                     /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(7616..7634)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7630..7677
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        7737..7841
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7842..7910,7911..8305,1..397)
                     /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(8060..8079)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
       61 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
      121 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
      181 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
      241 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
      301 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
      361 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
      421 tatactttag attgatttac cccggttgat aatcagaaaa gccccaaaaa caggaagatt
      481 gtataagcaa atatttaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt
      541 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
      601 aaagaatagc ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
      661 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
      721 cgtgaaccat cacccaaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
      781 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
      841 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
      901 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtaaaaggat
      961 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     1021 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     1081 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     1141 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     1201 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     1261 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     1321 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     1381 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     1441 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     1501 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     1561 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     1621 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     1681 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     1741 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     1801 gcgcagcgag tcagtgagcg aggaagctat ggtgcactct cagtacaatc tgctctgatg
     1861 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     1921 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     1981 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     2041 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgca gcgattcaca
     2101 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     2161 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc
     2221 cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct
     2281 cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca
     2341 actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagccgaa
     2401 cgccagcaag acgtagccca gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc
     2461 gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc
     2521 gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa
     2581 aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat
     2641 aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     2701 tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc
     2761 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     2821 cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca
     2881 ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca
     2941 agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg
     3001 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa
     3061 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
     3121 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
     3181 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
     3241 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca
     3301 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt
     3361 catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg
     3421 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa
     3481 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga
     3541 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt
     3601 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa
     3661 tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct
     3721 ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca
     3781 ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta
     3841 ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc
     3901 gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc
     3961 tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat
     4021 ggtgcatgcc ggcatgccgc cctttcgtct tcaagaatta attcccaatt ccccaggcat
     4081 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     4141 gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa
     4201 cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggaatta attccccagg
     4261 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     4321 tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag
     4381 caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggaa ttaattcccc
     4441 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     4501 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg
     4561 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gaattaattc
     4621 cccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     4681 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt
     4741 gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggaattaa
     4801 ttccccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     4861 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac
     4921 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggaat
     4981 tggggatcgg aattaattcc cggtttaaac cggggatctc gatcccgcga aattaatacg
     5041 actcactata ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa
     5101 ctttaagaag gagatataca tatgatggag cggaagaggt gggagtgccc ggcgctcccg
     5161 cagggctggg aaagggaaga agtgcccagg aggtcggggc tgtcggccgg ccacagggat
     5221 gtcttttact atagccccag cgggaagaag ttccgcagca agccacaact ggcacgttac
     5281 ctgggcggat ccatggacct cagcaccttc gacttccgca ccggaaagat gttgatgaac
     5341 aagatgaata agagtcgcca gcgtgtgcgc tatgattctt ccaaccaggt caagggcaag
     5401 cctgacctga acaccgcgct gcctgtacgg cagactgcat ccatcttcaa gcaaccggtg
     5461 accaagatca ccaaccaccc cagcaacaag gtcaagagcg acccgcagaa ggcagtggac
     5521 cagccgaggc agcttttctg ggagaagaag ctaagtggat tgagtgcctt tgacattgca
     5581 gaagaactgg tcaggaccat ggacttgccc aagggcctgc agggagtggg ccctggctgt
     5641 acagatgaga cgctgctgtc agccattgcg agtgctctac acaccagcac cctgcccatt
     5701 acaggccagc tctctgcagc cgtggagaag aaccctggtg tgtggctgaa cactgcacag
     5761 ccactgtgca aagccttcat ggtgacagat gacgacatca ggaagcagga ggagctggta
     5821 cagcaggtac ggaagcgcct ggaggaggca ctgatggccg acatgctagc tcatgtggag
     5881 gagcttgccc gagacgggga ggcaccactg gacaaggcct gtgcagagga ggaagaggag
     5941 gaggaagagg aggaggaaga gccggagcca gagcgagtgg aattcctcga gggctcttcc
     6001 tgctttgcca agggtaccaa tgttttaatg gcggatgggt ctattgaatg tattgaaaac
     6061 attgaggttg gtaataaggt catgggtaaa gatggcagac ctcgtgaggt aattaaattg
     6121 cccagaggaa gagaaactat gtacagcgtc gtgcagaaaa gtcagcacag agcccacaaa
     6181 agtgactcaa gtcgtgaagt gccagaatta ctcaagttta cgtgtaatgc gacccatgag
     6241 ttggttgtta gaacacctcg tagtgtccgc cgtttgtctc gtaccattaa gggtgtcgaa
     6301 tattttgaag ttattacttt tgagatgggc caaaagaaag cccccgacgg tagaattgtt
     6361 gagcttgtca aggaagtttc aaagagctac ccaatatctg aggggcctga gagagccaac
     6421 gaattagtag aatcctatag aaaggcttca aataaagctt attttgagtg gactattgag
     6481 gccagagatc tttctctgtt gggttcccat gttcgtaaag ctacctacca gacttacgct
     6541 ccaattcttt atgagaatga ccactttttc gactacatgc aaaaaagtaa gtttcatctc
     6601 accattgaag gtccaaaagt acttgcttat ttacttggtt tatggattgg tgatggattg
     6661 tctgacaggg caactttttc ggttgattcc agagatactt ctttgatgga acgtgttact
     6721 gaatatgctg aaaagttgaa tttgtgcgcc gagtataagg acagaaaaga accacaagtt
     6781 gccaaaactg ttaatttgta ctctaaagtt gtcagaggta atggtattcg caataatctt
     6841 aatactgaga atccattatg ggacgctatt gttggcttag gattcttgaa ggacggtgtc
     6901 aaaaatattc cttctttctt gtctacggac aatatcggta ctcgtgaaac atttcttgct
     6961 ggtctaattg attctgatgg ctatgttact gatgagcatg gtattaaagc aacaataaag
     7021 acaattcata cttctgtcag agatggtttg gtttcccttg ctcgttcttt aggcttagta
     7081 gtctcggtta acgcagaacc tgctaaggtt gacatgaatg tcaccaaaca taaaattagt
     7141 tatgctattt atatgtctgg tggagatgtt ttgcttaacg ttctttcgaa gtgtgccggc
     7201 tctaaaaaat tcaggcctgc tcccgccgct gcttttgcac gtgagtgccg cggattttat
     7261 ttcgagttac aagaattgaa ggaagacgat tattatggga ttactttatc tgatgattct
     7321 gatcatcagt ttttgcttgg atcccaggtt gtcgtccatg catgcggtgg cctgaccggt
     7381 ctgaactcag gcctcacgac aaatcctggt gtatccgctt ggcaggtcaa cacagcttat
     7441 actgcgggac aattggtcac atataacggc aagacgtata aatgtttgca gccccacacc
     7501 tccttggcag gatgggaacc atccaacgtt cctgccttgt ggcagcttca atgactgcag
     7561 gaaggggatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     7621 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
     7681 aaaggaggaa ctatatccgg ataactacgt caggtggcac ttttcgggga aatgtgcgcg
     7741 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     7801 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     7861 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     7921 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     7981 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt ttcccaatga
     8041 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     8101 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     8161 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     8221 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     8281 ccgctttttt gcacaacatg gggga
//
