LOCUS       sequence-460766-        7990 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Bacterial expression plasmid of MBD2-C (C-terminus of MBD2: aa 
            236-414). C-terminal tagged to intein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7990)
  TITLE     MBD2 modulates heterchromatin architecture and composition
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7990)
  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            "Unpublished"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 pc4793
FEATURES             Location/Qualifiers
     source          1..7990
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(81..100)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      700..1213
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(787..806)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     997..1018
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(1255..2046,2047..2115))
                     /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     1878..1897
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2116..2220)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(2280..2327)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2323..2341
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(2406..2561)
                     /codon_start=1
                     /gene="Bacillus circulans chiA"
                     /product="chitin binding domain from chitinase A1 (Watanabe
                     et al., 1994)"
                     /label=CBD
                     /translation="TTNPGVSAWQVNTAYTAGQLVTYNGKTYKCLQPHTSLAGWEPSNV
                     PALWQLQ"
     CDS             complement(2671..2682)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(3138..3956)
                     /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"
     RBS             4528..4550
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4565..4589
                     /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)."
     primer_bind     complement(4589..4608)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4590..4608)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(4743..4829)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(4926..5012)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(5109..5195)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(5292..5378)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(5475..5561)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        5714..5791
                     /gene="lacI"
                     /label=lacI promoter
     CDS             5792..6874
                     /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     complement(5811..5830)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             7431..7622
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(7616..7638)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7779..7798
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7820..7837)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
       61 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      121 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      181 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      241 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      301 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      361 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      421 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
      481 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
      541 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
      601 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
      661 catgagatta tcaaaaagga tcttcaccta gatcctttta cgcgccctgt agcggcgcat
      721 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
      781 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
      841 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
      901 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
      961 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     1021 caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     1081 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     1141 taacgtttac aatttaaata tttgcttata caatcttcct gtttttgggg cttttctgat
     1201 tatcaaccgg ggtaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     1261 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1321 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     1381 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     1441 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1501 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     1561 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     1621 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     1681 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     1741 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     1801 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     1861 cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     1921 tgggaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     1981 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     2041 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     2101 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     2161 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     2221 cacatttccc cgaaaagtgc cacctgacgt agttatccgg atatagttcc tcctttcagc
     2281 aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctagtta ttgctcagcg
     2341 gtggcagcag ccaactcagc ttcctttcgg gctttgttag cagccggatc cccttcctgc
     2401 agtcattgaa gctgccacaa ggcaggaacg ttggatggtt cccatcctgc caaggaggtg
     2461 tggggctgca aacatttata cgtcttgccg ttatatgtga ccaattgtcc cgcagtataa
     2521 gctgtgttga cctgccaagc ggatacacca ggatttgtcg tgaggcctga gttcagaccg
     2581 gtcaggccac cgcatgcatg gacgacaacc tgggatccaa gcaaaaactg atgatcagaa
     2641 tcatcagata aagtaatccc ataataatcg tcttccttca attcttgtaa ctcgaaataa
     2701 aatccgcggc actcacgtgc aaaagcagcg gcgggagcag gcctgaattt tttagagccg
     2761 gcacacttcg aaagaacgtt aagcaaaaca tctccaccag acatataaat agcataacta
     2821 attttatgtt tggtgacatt catgtcaacc ttagcaggtt ctgcgttaac cgagactact
     2881 aagcctaaag aacgagcaag ggaaaccaaa ccatctctga cagaagtatg aattgtcttt
     2941 attgttgctt taataccatg ctcatcagta acatagccat cagaatcaat tagaccagca
     3001 agaaatgttt cacgagtacc gatattgtcc gtagacaaga aagaaggaat atttttgaca
     3061 ccgtccttca agaatcctaa gccaacaata gcgtcccata atggattctc agtattaaga
     3121 ttattgcgaa taccattacc tctgacaact ttagagtaca aattaacagt tttggcaact
     3181 tgtggttctt ttctgtcctt atactcggcg cacaaattca acttttcagc atattcagta
     3241 acacgttcca tcaaagaagt atctctggaa tcaaccgaaa aagttgccct gtcagacaat
     3301 ccatcaccaa tccataaacc aagtaaataa gcaagtactt ttggaccttc aatggtgaga
     3361 tgaaacttac ttttttgcat gtagtcgaaa aagtggtcat tctcataaag aattggagcg
     3421 taagtctggt aggtagcttt acgaacatgg gaacccaaca gagaaagatc tctggcctca
     3481 atagtccact caaaataagc tttatttgaa gcctttctat aggattctac taattcgttg
     3541 gctctctcag gcccctcaga tattgggtag ctctttgaaa cttccttgac aagctcaaca
     3601 attctaccgt cgggggcttt cttttggccc atctcaaaag taataacttc aaaatattcg
     3661 acacccttaa tggtacgaga caaacggcgg acactacgag gtgttctaac aaccaactca
     3721 tgggtcgcat tacacgtaaa cttgagtaat tctggcactt cacgacttga gtcacttttg
     3781 tgggctctgt gctgactttt ctgcacgacg ctgtacatag tttctcttcc tctgggcaat
     3841 ttaattacct cacgaggtct gccatcttta cccatgacct tattaccaac ctcaatgttt
     3901 tcaatacatt caatagaccc atccgccatt aaaacattgg tacccttggc aaagcaggaa
     3961 gagccctcga ggaattccgc ctcatctcca tcgtccatgt caatgtctac ttcctccgtg
     4021 tccgcagccc gggacaggat gtcggccatc agtgcctcct ccagtttctt gcgtacttgt
     4081 tggactcgct cttcctgttt cctaatgtct tcatctgtaa caatgaaagc tttgcagagg
     4141 ggttgagatg tgttaagcca aacagcaggg ttcttttcca cggcagcaga gacttgtcct
     4201 gtgatgggcg cagagcttgt gtgtaaagca ctggccacag cagacagaag ggtctcgtca
     4261 ttgctacctg gaccgactcc ttgaagacct ttaggtagct ccatggtttt tataatttgt
     4321 tctgttacat ctgatgcgct aagtccttgt agcctcttct cccagaaaag ctgacgtggt
     4381 tgttcattca tccgctgggg gtctgacttc accttattgc tcgggtggtt cgtgaatttg
     4441 gttactggtt gcttgaaaat tgatgcagtt tgtctaattg gcaatgttgt gttcaggtct
     4501 ggtttaccct tgttcatcat atgtatatct ccttcttaaa gttaaacaaa attatttcta
     4561 gaggggaatt gttatccgct cacaattccc ctatagtgag tcgtattaat ttcgcgggat
     4621 cgagatcccc ggtttaaacc gggaattaat tccgatcccc aattcctggc agtttatggc
     4681 gggcgtcctg cccgccaccc tccgggccgt tgcttcgcaa cgttcaaatc cgctcccggc
     4741 ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag
     4801 tctttcgact gagcctttcg ttttatttga tgcctgggga attaattcct ggcagtttat
     4861 ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttcg caacgttcaa atccgctccc
     4921 ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc
     4981 cagtctttcg actgagcctt tcgttttatt tgatgcctgg ggaattaatt cctggcagtt
     5041 tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcgcaacgtt caaatccgct
     5101 cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag
     5161 gcccagtctt tcgactgagc ctttcgtttt atttgatgcc tggggaatta attcctggca
     5221 gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcgcaac gttcaaatcc
     5281 gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga
     5341 aaggcccagt ctttcgactg agcctttcgt tttatttgat gcctggggaa ttaattcctg
     5401 gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcgc aacgttcaaa
     5461 tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa
     5521 cgaaaggccc agtctttcga ctgagccttt cgttttattt gatgcctggg gaattgggaa
     5581 ttaattcttg aagacgaaag ggcggcatgc cggcatgcac cattccttgc ggcggcggtg
     5641 ctcaacggcc tcaacctact actgggctgc ttcctaatgc aggagtcgca taagggagag
     5701 cgtcgagatc ccggacacca tcgaatggcg caaaaccttt cgcggtatgg catgatagcg
     5761 cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc
     5821 agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt
     5881 ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa
     5941 ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag
     6001 tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact
     6061 gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc
     6121 ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga
     6181 ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt
     6241 ctctgaccag acacccatca acagtattat tttctcccat gaagacggta cgcgactggg
     6301 cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag
     6361 ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat
     6421 tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat
     6481 gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc
     6541 gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt
     6601 agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa
     6661 acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg
     6721 ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct
     6781 ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     6841 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc
     6901 tcactcatta ggcaccggga tctcgaccga tgcccttgag agccttcaac ccagtcagct
     6961 ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca
     7021 tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat tttcggcgag gaccgctttc
     7081 gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg
     7141 ctcaagcctt cgtcactggt cccgccacca aacgtttcgg cgagaagcag gccattatcg
     7201 ccggcatggc ggccgacgcg ctgggctacg tcttgctggc gttcggctgg cattgaccct
     7261 gagtgatttt tctctggtcc cgccgcatcc ataccgccag ttgtttaccc tcacaacgtt
     7321 ccagtaaccg ggcatgttca tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca
     7381 tcggtatcat tacccccatg aacagaaatc ccccttacac ggaggcatca gtgaccaaac
     7441 aggaaaaaac cgcccttaac atggcccgct ttatcagaag ccagacatta acgcttctgg
     7501 agaaactcaa cgagctggac gcggatgaac aggcagacat ctgtgaatcg ctgcacgacc
     7561 acgctgatga gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct
     7621 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     7681 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt
     7741 cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact
     7801 gagagtgcac catagcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     7861 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     7921 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     7981 tgctggcgtt
//
