LOCUS       sequence-460761-        8611 bp    DNA     circular SYN 16-MAY-2025
DEFINITION  Bacterial expression vector of MBD2a lacking the coiled coil domain:
            aa 363-396..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8611)
  TITLE     MBD2 modulates heterchromatin architecture and composition
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8611)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pTYB1-MBD2a∆CC pc5081
FEATURES             Location/Qualifiers
     source          1..8611
                     /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             5245..6063
                     /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             6519..6530
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             6640..6795
                     /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(6860..6878)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6874..6921
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        6981..7085
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7086..7154,7155..7946)
                     /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(7304..7323)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7988..8501)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8183..8204)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8395..8414
                     /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 tgtcaaaggc ctacgtcgac
     4081 gccatgcgcg cgcacccggg gggaggccgc tgctgcccgg agcaggagga gggggagagc
     4141 gcggcgggcg gcagcggcgc tggcggcgac tccgccatag agcagggggg ccagggcagc
     4201 gcgctcgctc cgtccccggt gagcggcgtg cgcagggaag gcgctcgggg cggcggccgt
     4261 ggccgggggc ggtggaagca ggcggcccgg ggcggcggcg tctgtggccg tggccgtggc
     4321 cgtggccggg gtcggggccg tggccggggc cggggccggg gccgcggccg tccccagagt
     4381 ggcggcagcg gccttggcgg cgacggcggc ggcggcgcgg gcggctgcgg cgtcggcagc
     4441 ggtggcggcg tcgccccccg gcgggatcct gtccctttcc cgtcggggag ctcggggccg
     4501 gggcccaggg gaccccgggc cacggagagc gggaagagga tggactgccc ggccctcccc
     4561 cccggatgga agaaggagga agtgatccga aaatcagggc tcagtgctgg caagagcgat
     4621 gtctactact tcagtccaag tggtaagaag ttcagaagta aacctcagct ggcaagatac
     4681 ctgggaaatg ctgttgacct tagcagtttt gacttcagga ccggcaagat gatgcctagt
     4741 aaattacaga agaacaagca gagactccgg aatgaccccc tcaatcagaa caagggtaaa
     4801 ccagacctga acacaacatt gccaattaga caaactgcat caattttcaa gcaaccagta
     4861 accaaattca cgaaccaccc gagcaataag gtgaagtcag acccccagcg gatgaatgaa
     4921 caaccacgtc agcttttctg ggagaagagg ctacaaggac ttagcgcatc agatgtaaca
     4981 gaacaaatta taaaaaccat ggagctacct aaaggtcttc aaggagtcgg tccaggtagc
     5041 aatgacgaga cccttctgtc tgctgtggcc agtgctttac acacaagctc tgcgcccatc
     5101 acaggacaag tctctgctgc cgtggaaaag aaccctgctg tttggcttaa cacatctcaa
     5161 cccctctgcc gggctgcgga cacggaggaa gtagacattg acatggacga tggagatgag
     5221 gcggaattcc tcgagggctc ttcctgcttt gccaagggta ccaatgtttt aatggcggat
     5281 gggtctattg aatgtattga aaacattgag gttggtaata aggtcatggg taaagatggc
     5341 agacctcgtg aggtaattaa attgcccaga ggaagagaaa ctatgtacag cgtcgtgcag
     5401 aaaagtcagc acagagccca caaaagtgac tcaagtcgtg aagtgccaga attactcaag
     5461 tttacgtgta atgcgaccca tgagttggtt gttagaacac ctcgtagtgt ccgccgtttg
     5521 tctcgtacca ttaagggtgt cgaatatttt gaagttatta cttttgagat gggccaaaag
     5581 aaagcccccg acggtagaat tgttgagctt gtcaaggaag tttcaaagag ctacccaata
     5641 tctgaggggc ctgagagagc caacgaatta gtagaatcct atagaaaggc ttcaaataaa
     5701 gcttattttg agtggactat tgaggccaga gatctttctc tgttgggttc ccatgttcgt
     5761 aaagctacct accagactta cgctccaatt ctttatgaga atgaccactt tttcgactac
     5821 atgcaaaaaa gtaagtttca tctcaccatt gaaggtccaa aagtacttgc ttatttactt
     5881 ggtttatgga ttggtgatgg attgtctgac agggcaactt tttcggttga ttccagagat
     5941 acttctttga tggaacgtgt tactgaatat gctgaaaagt tgaatttgtg cgccgagtat
     6001 aaggacagaa aagaaccaca agttgccaaa actgttaatt tgtactctaa agttgtcaga
     6061 ggtaatggta ttcgcaataa tcttaatact gagaatccat tatgggacgc tattgttggc
     6121 ttaggattct tgaaggacgg tgtcaaaaat attccttctt tcttgtctac ggacaatatc
     6181 ggtactcgtg aaacatttct tgctggtcta attgattctg atggctatgt tactgatgag
     6241 catggtatta aagcaacaat aaagacaatt catacttctg tcagagatgg tttggtttcc
     6301 cttgctcgtt ctttaggctt agtagtctcg gttaacgcag aacctgctaa ggttgacatg
     6361 aatgtcacca aacataaaat tagttatgct atttatatgt ctggtggaga tgttttgctt
     6421 aacgttcttt cgaagtgtgc cggctctaaa aaattcaggc ctgctcccgc cgctgctttt
     6481 gcacgtgagt gccgcggatt ttatttcgag ttacaagaat tgaaggaaga cgattattat
     6541 gggattactt tatctgatga ttctgatcat cagtttttgc ttggatccca ggttgtcgtc
     6601 catgcatgcg gtggcctgac cggtctgaac tcaggcctca cgacaaatcc tggtgtatcc
     6661 gcttggcagg tcaacacagc ttatactgcg ggacaattgg tcacatataa cggcaagacg
     6721 tataaatgtt tgcagcccca cacctccttg gcaggatggg aaccatccaa cgttcctgcc
     6781 ttgtggcagc ttcaatgact gcaggaaggg gatccggctg ctaacaaagc ccgaaaggaa
     6841 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     6901 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggataact acgtcaggtg
     6961 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     7021 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     7081 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     7141 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     7201 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     7261 gccccgaaga acgtttccca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     7321 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     7381 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     7441 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     7501 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     7561 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     7621 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     7681 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     7741 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     7801 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     7861 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     7921 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     7981 ttgatttacc ccggttgata atcagaaaag ccccaaaaac aggaagattg tataagcaaa
     8041 tatttaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     8101 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagcc
     8161 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     8221 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     8281 acccaaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     8341 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     8401 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     8461 caccacaccc gccgcgctta atgcgccgct acagggcgcg taaaaggatc taggtgaaga
     8521 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     8581 cagaccccgt agaaaagatc aaaggatctt c
//
