LOCUS       sequence-236188-        5994 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression Brg1 amino acids 1446-1647 in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5994)
  AUTHORS   Zhou J, Zhang M, Fang H, El-Mounayri O, Rodenberg JM, Imbalzano AN, 
            Herring BP
  TITLE     The SWI/SNF chromatin remodeling complex regulates myocardin-induced
            smooth muscle-specific gene expression.
  JOURNAL   Arterioscler Thromb Vasc Biol. 2009 Jun;29(6):921-8. doi: 
            10.1161/ATVBAHA.109.187229. Epub 2009 Apr 2.
  PUBMED    19342595
REFERENCE   2  (bases 1 to 5994)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1161/ATVBAHA.109.187229"; journalName: "Arterioscler Thromb Vasc
            Biol"; date: "2009-06"; volume: "29"; issue: "6"; pages: "921-8"
COMMENT     Sequence Label: Brg1 Del4 pET28C
FEATURES             Location/Qualifiers
     source          1..5994
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             19..36
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             46..63
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             67..99
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             742..765
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             778..795
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(848..866)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      862..909
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      946..1401
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1033..1052)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1243..1264
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1493..2308)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1493..2308)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2216..2235
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2300..2319
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2430..3018
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2919..2938
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3172..3189
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3204..3343
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3269..3288)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3429..3451
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3445..3636)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4445..5527)
                     /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     5489..5508
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5528..5605)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5811..5830
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5914..5933
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5914..5932
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5933..5957
                     /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             5972..5994
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 tataccatgg gcagcagcca tcatcatcat catcacagca gcggcctggt gccgcgcggc
       61 agccatatgg ctagcatgac tggtggacag caaatgggtc ggatccgaat tcgagctccg
      121 tcgacaagct tgcggccgcc tgccgagaaa ctctccccta acccacccaa cctcaccaag
      181 aagatgaaga agattgtgga tgccgtgatc aagtacaagg acagcagcag tggacgtcag
      241 ctcagcgagg tcttcatcca gctgccctcg cgaaaggagc tgcccgagta ctacgagctc
      301 atccgcaagc ccgtggactt caagaagata aaggagcgca ttcgcaacca caagtaccgc
      361 agcctcaacg acctagagaa ggacgtcatg ctcctgtgcc agaacgcaca gaccttcaac
      421 ctggagggct ccctgatcta tgaagactcc atcgtcttgc agtcggtctt caccagcgtg
      481 cggcagaaaa tcgagaagga ggatgacagt gaaggcgagg agagtgagga ggaggaagag
      541 ggcgaggagg aaggctccga atccgaatct cggtccgtca aagtgaagat caagcttggc
      601 cggaaggaga aggcacagga ccagctgaag ggcggccggc ggcggccgag ccgagggtcc
      661 cgagccaagc cggtcgtgag tgacgatgac agtgaggagg aacaagagga ggaccgctca
      721 ggaagtggca gcgaagaaga cgactacaag gacgacgatg acaaatgata agtcgagcac
      781 caccaccacc accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct
      841 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
      901 ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg
      961 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     1021 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     1081 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     1141 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     1201 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     1261 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     1321 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     1381 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
     1441 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgaattaat tcttagaaaa
     1501 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt
     1561 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
     1621 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
     1681 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg
     1741 gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac
     1801 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag
     1861 cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc
     1921 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta
     1981 atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag
     2041 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
     2101 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
     2161 gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc
     2221 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctagagc
     2281 aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag
     2341 acagttttat tgttcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2401 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2461 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2521 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     2581 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2641 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2701 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2761 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2821 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2881 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     2941 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3001 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3061 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3121 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3181 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     3241 atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     3301 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     3361 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     3421 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     3481 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     3541 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     3601 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt
     3661 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
     3721 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
     3781 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt
     3841 aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca
     3901 gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt
     3961 tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg
     4021 tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag
     4081 gagcacgatc atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc
     4141 gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa
     4201 taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat
     4261 gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag
     4321 tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct
     4381 caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt
     4441 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4501 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca
     4561 gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc
     4621 ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga
     4681 tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc
     4741 gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca
     4801 gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca
     4861 tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt
     4921 tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg
     4981 cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat
     5041 gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa
     5101 cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga
     5161 tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc
     5221 cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa
     5281 tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca
     5341 gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg
     5401 ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca
     5461 cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg
     5521 gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa
     5581 aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc
     5641 attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt
     5701 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
     5761 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
     5821 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
     5881 gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt
     5941 gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag gaga
//
