LOCUS       sequence-456555-        5742 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression for structure determination; may not be full 
            ORF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5742)
  TITLE     SGC+year
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 5742)
  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"; journalName: 
            "unpublished"
COMMENT     Sequence Label: BRD4
FEATURES             Location/Qualifiers
     source          1..5742
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             92..109
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             134..154
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             607..624
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(677..695)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      691..738
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      775..1230
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(862..881)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1072..1093
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1322..2137)
                     /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(1322..2137)
                     /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     2045..2064
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2129..2148
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2259..2847
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2748..2767
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3001..3018
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3033..3172
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3098..3117)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3258..3280
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3274..3465)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4274..5356)
                     /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     5318..5337
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5357..5434)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5640..5659
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gcaccatcat catcatcatt cttctggtgt
      121 agatctgggt accgagaacc tgtacttcca atccatgaag gacgtgcccg actctcagca
      181 gcacccagca ccagagaaga gcagcaaggt ctcggagcag ctcaagtgct gcagcggcat
      241 cctcaaggag atgtttgcca agaagcacgc cgcctacgcc tggcccttct acaagcctgt
      301 ggacgtggag gcactgggcc tacacgacta ctgtgacatc atcaagcacc ccatggacat
      361 gagcacaatc aagtctaaac tggaggcccg tgagtaccgt gatgctcagg agtttggtgc
      421 tgacgtccga ttgatgttct ccaactgcta taagtacaac cctcctgacc atgaggtggt
      481 ggccatggcc cgcaagctcc aggatgtgtt cgaaatgcgc tttgccaaga tgccggacga
      541 gtgacagtaa aggtggatac ggatccgaat tcgagctccg tcgacaagct tgcggccgca
      601 ctcgagcacc accaccacca ccactgagat ccggctgcta acaaagcccg aaaggaagct
      661 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
      721 gtcttgaggg gttttttgct gaaaggagga actatatccg gattggcgaa tgggacgcgc
      781 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
      841 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
      901 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
      961 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     1021 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     1081 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     1141 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     1201 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     1261 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgaattaatt
     1321 cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa
     1381 taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc
     1441 ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac
     1501 ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga
     1561 ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc
     1621 agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt
     1681 gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg
     1741 aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat
     1801 attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat
     1861 catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt
     1921 ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa
     1981 acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga
     2041 cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg
     2101 gcctagagca agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta
     2161 tgtaagcaga cagttttatt gttcatgacc aaaatccctt aacgtgagtt ttcgttccac
     2221 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2281 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2341 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     2401 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2461 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2521 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2581 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2641 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2701 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2761 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2821 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2881 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     2941 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     3001 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     3061 ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat
     3121 agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca
     3181 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     3241 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     3301 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc
     3361 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct
     3421 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta
     3481 agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat
     3541 acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc
     3601 ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa
     3661 tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat
     3721 aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac
     3781 cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc
     3841 gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct
     3901 caacgacagg agcacgatca tgcgcacccg tggggccgcc atgccggcga taatggcctg
     3961 cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa
     4021 gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc
     4081 gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac
     4141 agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt
     4201 gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt
     4261 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4321 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc
     4381 ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt
     4441 gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta
     4501 acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg
     4561 caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt
     4621 tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa
     4681 aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag
     4741 tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg
     4801 ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac
     4861 cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata
     4921 acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat
     4981 agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg
     5041 cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc
     5101 gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa
     5161 cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat
     5221 tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct
     5281 ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata
     5341 acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca
     5401 taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc
     5461 gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca
     5521 aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc
     5581 atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg
     5641 gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg
     5701 atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa at
//
