LOCUS       sequence-177425-        7002 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human BROX residues 1-401 in bacteria; internal ID 
            WISP11-297.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7002)
  AUTHORS   Zhai Q, Landesman MB, Robinson H, Sundquist WI, Hill CP
  TITLE     Structure of the Bro1 domain protein BROX and functional analyses of
            the ALIX Bro1 domain in HIV-1 budding.
  JOURNAL   PLoS One. 2011;6(12):e27466. doi: 10.1371/journal.pone.0027466. Epub
            2011 Dec 5.
  PUBMED    22162750
REFERENCE   2  (bases 1 to 7002)
  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: "PLoS One.";
            date: "2011"; volume: "6(12):e27466. doi"; pages: " 
            10.1371/journal.pone.0027466. Epub 2011 Dec 5"
COMMENT     Sequence Label: pet151D-TOPO-BROX 1-401
FEATURES             Location/Qualifiers
     source          1..7002
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     106..125
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     209..228
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        209..227
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    228..252
                     /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             267..289
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             300..317
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             318..359
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             360..380
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1703..1721)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1717..1764
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2002..2020)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2088..2192
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2193..2261,2262..3053)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2411..2430)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3224..3812
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3713..3732
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3966..3983
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3998..4137
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4063..4082)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4223..4245
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4239..4430)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5742..6824)
                     /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     6786..6805
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6825..6902)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
       61 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
      121 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
      181 gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg
      241 gataacaatt cccctctaga aataattttg tttaacttta agaaggagat atacatatgc
      301 atcatcacca tcaccatggt aagcctatcc ctaaccctct cctcggtctc gattctacgg
      361 aaaacctgta ttttcaggga attgatccct tcaccatgac ccattggttt cataggaacc
      421 cattaaaagc cacagctcct gtgtctttta attactatgg tgtagtcact ggcccttctg
      481 cttcaaaaat atgcaatgac ttgaggtcat ccagggcacg actccttgaa ctgttcactg
      541 atttgagctg taatccagaa atgatgaaga atgcagcaga ttcatatttc tcacttttac
      601 aaggtttcat aaattctttg gatgaatcta cccaagaaag caagttacga tatattcaaa
      661 atttcaagtg gactgataca ttgcaaggac aggttccaag tgcccagcag gatgctgttt
      721 ttgaattaat ttccatggga tttaatgtag ctttatggta taccaaatat gcttcaagac
      781 tggctggaaa agaaaatata acagaagatg aagcaaaaga agttcatcga agcctaaaga
      841 ttgcagctgg gatttttaaa catttaaagg aaagtcatct cccaaaactc attacacctg
      901 cggaaaaagg aagagattta gagtcacgac tcatagaagc atacgttatt caatgtcagg
      961 ctgaagctca agaagtaaca attgctcgag caattgaact aaaacatgct cctggactaa
     1021 ttgctgcact ggcgtatgaa acagccaatt tctatcaaaa agctgatcat actttatcca
     1081 gtttggagcc tgcatattct gccaaatgga gaaaatatct tcacttgaag atgtgtttct
     1141 acacagctta tgcttactgt taccatggtg agactttatt ggctagtgat aaatgcggtg
     1201 aagcaatcag gtctctccaa gaagcagaaa aattgtatgc aaaggcagaa gcactgtgta
     1261 aagaatatgg agaaaccaaa ggacctggac caacagtcaa accttcagga catctgttct
     1321 ttaggaaact tggaaacctt gtgaagaaca ccctagaaaa atgtcagaga gaaaatggat
     1381 ttatttactt tcaaaaaatt ccaacagaag ccccacagct ggaactcaaa gcaaattatg
     1441 gtctcgtaga gcctatacct ttcgaatttc ctcctacaag tgttcagtgg acaccagaaa
     1501 cattggctgc atttgatctc accaaaagac ccaaggatga cagtactaaa cccaaaccag
     1561 aagaagaagt gaaacctgtg aaagaaccag acatcaaata acctcaaaag gacactgggt
     1621 gctacatctc ctaacatatg aagggcgagc tcagatccgg ctgctaacaa agcccgaaag
     1681 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
     1741 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggata tcccgcaaga
     1801 ggcccggcag taccggcata accaagccta tgcctacagc atccagggtg acggtgccga
     1861 ggatgacgat gagcgcattg ttagatttca tacacggtgc ctgactgcgt tagcaattta
     1921 actgtgataa actaccgcat taaagctagc ttatcgatga taagctgtca aacatgagaa
     1981 ttaattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     2041 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2101 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2161 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     2221 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     2281 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     2341 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     2401 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     2461 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     2521 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     2581 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     2641 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     2701 ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca
     2761 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     2821 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     2881 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     2941 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3001 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     3061 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     3121 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     3181 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3241 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3301 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3361 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3421 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3481 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3541 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3601 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3661 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3721 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3781 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3841 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     3901 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     3961 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     4021 cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc
     4081 cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc
     4141 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     4201 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     4261 ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag
     4321 atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg
     4381 cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc
     4441 gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc
     4501 acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa
     4561 ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc
     4621 gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg
     4681 aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg
     4741 aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt
     4801 cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg
     4861 gtcctcaacg acaggagcac gatcatgcgc acccgtggcc aggacccaac gctgcccgag
     4921 atgcgccgcg tgcggctgct ggagatggcg gacgcgatgg atatgttctg ccaagggttg
     4981 gtttgcgcat tcacagttct ccgcaagaat tgattggctc caattcttgg agtggtgaat
     5041 ccgttagcga ggtgccgccg gcttccattc aggtcgaggt ggcccggctc catgcaccgc
     5101 gacgcaacgc ggggaggcag acaaggtata gggcggcgcc tacaatccat gccaacccgt
     5161 tccatgtgct cgccgaggcg gcataaatcg ccgtgacgat cagcggtcca atgatcgaag
     5221 ttaggctggt aagagccgcg agcgatcctt gaagctgtcc ctgatggtcg tcatctacct
     5281 gcctggacag catggcctgc aacgcgggca tcccgatgcc gccggaagcg agaagaatca
     5341 taatggggaa ggccatccag cctcgcgtcg cgaacgccag caagacgtag cccagcgcgt
     5401 cggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag
     5461 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca
     5521 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct
     5581 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc
     5641 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg
     5701 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
     5761 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5821 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg
     5881 cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc
     5941 aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta
     6001 tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg
     6061 cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc
     6121 tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt
     6181 tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga
     6241 cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg
     6301 accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg
     6361 ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca
     6421 gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg
     6481 agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc
     6541 accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc
     6601 gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt
     6661 tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc
     6721 cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag
     6781 acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat
     6841 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg
     6901 tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag
     6961 gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tg
//
