LOCUS       sequence-459932-        7507 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Bacterial expression vector of MBD2-MBDTRD domains (aa 153-235) 
            C-terminal tagged to the intein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7507)
  TITLE     MBD2 modulates heterchromatin architecture and composition
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7507)
  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            "Unpublished"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-MBDTRD pc4791
FEATURES             Location/Qualifiers
     source          1..7507
                     /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             4141..4959
                     /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             5415..5426
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             5536..5691
                     /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(5756..5774)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5770..5817
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        5877..5981
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5982..6050,6051..6842)
                     /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(6200..6219)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6884..7397)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7079..7100)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7291..7310
                     /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 tcaatcagga gggctcttcc
     4141 tgctttgcca agggtaccaa tgttttaatg gcggatgggt ctattgaatg tattgaaaac
     4201 attgaggttg gtaataaggt catgggtaaa gatggcagac ctcgtgaggt aattaaattg
     4261 cccagaggaa gagaaactat gtacagcgtc gtgcagaaaa gtcagcacag agcccacaaa
     4321 agtgactcaa gtcgtgaagt gccagaatta ctcaagttta cgtgtaatgc gacccatgag
     4381 ttggttgtta gaacacctcg tagtgtccgc cgtttgtctc gtaccattaa gggtgtcgaa
     4441 tattttgaag ttattacttt tgagatgggc caaaagaaag cccccgacgg tagaattgtt
     4501 gagcttgtca aggaagtttc aaagagctac ccaatatctg aggggcctga gagagccaac
     4561 gaattagtag aatcctatag aaaggcttca aataaagctt attttgagtg gactattgag
     4621 gccagagatc tttctctgtt gggttcccat gttcgtaaag ctacctacca gacttacgct
     4681 ccaattcttt atgagaatga ccactttttc gactacatgc aaaaaagtaa gtttcatctc
     4741 accattgaag gtccaaaagt acttgcttat ttacttggtt tatggattgg tgatggattg
     4801 tctgacaggg caactttttc ggttgattcc agagatactt ctttgatgga acgtgttact
     4861 gaatatgctg aaaagttgaa tttgtgcgcc gagtataagg acagaaaaga accacaagtt
     4921 gccaaaactg ttaatttgta ctctaaagtt gtcagaggta atggtattcg caataatctt
     4981 aatactgaga atccattatg ggacgctatt gttggcttag gattcttgaa ggacggtgtc
     5041 aaaaatattc cttctttctt gtctacggac aatatcggta ctcgtgaaac atttcttgct
     5101 ggtctaattg attctgatgg ctatgttact gatgagcatg gtattaaagc aacaataaag
     5161 acaattcata cttctgtcag agatggtttg gtttcccttg ctcgttcttt aggcttagta
     5221 gtctcggtta acgcagaacc tgctaaggtt gacatgaatg tcaccaaaca taaaattagt
     5281 tatgctattt atatgtctgg tggagatgtt ttgcttaacg ttctttcgaa gtgtgccggc
     5341 tctaaaaaat tcaggcctgc tcccgccgct gcttttgcac gtgagtgccg cggattttat
     5401 ttcgagttac aagaattgaa ggaagacgat tattatggga ttactttatc tgatgattct
     5461 gatcatcagt ttttgcttgg atcccaggtt gtcgtccatg catgcggtgg cctgaccggt
     5521 ctgaactcag gcctcacgac aaatcctggt gtatccgctt ggcaggtcaa cacagcttat
     5581 actgcgggac aattggtcac atataacggc aagacgtata aatgtttgca gccccacacc
     5641 tccttggcag gatgggaacc atccaacgtt cctgccttgt ggcagcttca atgactgcag
     5701 gaaggggatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     5761 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
     5821 aaaggaggaa ctatatccgg ataactacgt caggtggcac ttttcgggga aatgtgcgcg
     5881 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     5941 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     6001 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     6061 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     6121 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt ttcccaatga
     6181 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     6241 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     6301 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     6361 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     6421 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     6481 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     6541 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     6601 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     6661 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     6721 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     6781 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6841 aactgtcaga ccaagtttac tcatatatac tttagattga tttaccccgg ttgataatca
     6901 gaaaagcccc aaaaacagga agattgtata agcaaatatt taaattgtaa acgttaatat
     6961 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     7021 aatcggcaaa atcccttata aatcaaaaga atagcccgag atagggttga gtgttgttcc
     7081 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     7141 cgtctatcag ggcgatggcc cactacgtga accatcaccc aaatcaagtt ttttggggtc
     7201 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     7261 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     7321 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     7381 gccgctacag ggcgcgtaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     7441 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     7501 gatcttc
//
