LOCUS       sequence-204178-        6684 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6684)
  AUTHORS   Brzovic PS, Lissounov A, Christensen DE, Hoyt DW, Klevit RE
  TITLE     A UbcH5/ubiquitin noncovalent complex is required for processive 
            BRCA1-directed ubiquitination.
  JOURNAL   Mol Cell. 2006 Mar 17. 21(6):873-80.
  PUBMED    16543155
REFERENCE   2  (bases 1 to 6684)
  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: "Mol Cell. 
            2006 Mar 17. 21(6):873-80."
COMMENT     Sequence Label: BRCA1 (1-304)
FEATURES             Location/Qualifiers
     source          1..6684
                     /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(1385..1403)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1399..1446
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1684..1702)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1770..1874
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1875..1943,1944..2735)
                     /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(2093..2112)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2906..3494
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3395..3414
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3648..3665
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3680..3819
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3745..3764)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3905..3927
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3921..4112)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5424..6506)
                     /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     6468..6487
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6507..6584)
                     /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 tcaccatgga tttatctgct cttcgcgttg
      421 aagaagtaca aaatgtcatt aatgctatgc agaaaatctt agagtgtccc atctgtctgg
      481 agttgatcaa ggaacctgtc tccacaaagt gtgaccacat attttgcaaa ttttgcatgc
      541 tgaaacttct caaccagaag aaagggcctt cacagtgtcc tttatgtaag aatgatataa
      601 ccaaaaggag cctacaagaa agtacgagat ttagtcaact tgttgaagag ctattgaaaa
      661 tcatttgtgc ttttcagctt gacacaggtt tggagtatgc aaacagctat aattttgcaa
      721 aaaaggaaaa taactctcct gaacatctaa aagatgaagt ttctatcatc caaagtatgg
      781 gctacagaaa ccgtgccaaa agacttctac agagtgaacc cgaaaatcct tccttgcagg
      841 aaaccagtct cagtgtccaa ctctctaacc ttggaactgt gagaactctg aggacaaagc
      901 agcggataca acctcaaaag acgtctgtct acattgaatt gggatctgat tcttctgaag
      961 ataccgttaa taaggcaact tattgcagtg tgggagatca agaattgtta caaatcaccc
     1021 ctcaaggaac cagggatgaa atcagtttgg attctgcaaa aaaggctgct tgtgaatttt
     1081 ctgagacgga tgtaacaaat actgaacatc atcaacccag taataatgat ttgaacacca
     1141 ctgagaagcg tgcagctgag aggcatccag aaaagtatca gggtagttct gtttcaaact
     1201 tgcatgtgga gccatgtggc acaaatactc atgccagctc attacagcat gagaacagca
     1261 gtttattact cactaaagac agaatgaatg tagaaaaggc tgaattctaa ctcgaggcgt
     1321 cgaagggcga gctcagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
     1381 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
     1441 tttttgctga aaggaggaac tatatccgga tatcccgcaa gaggcccggc agtaccggca
     1501 taaccaagcc tatgcctaca gcatccaggg tgacggtgcc gaggatgacg atgagcgcat
     1561 tgttagattt catacacggt gcctgactgc gttagcaatt taactgtgat aaactaccgc
     1621 attaaagcta gcttatcgat gataagctgt caaacatgag aattaattct tgaagacgaa
     1681 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     1741 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     1801 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     1861 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     1921 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     1981 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     2041 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     2101 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     2161 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     2221 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     2281 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     2341 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     2401 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     2461 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     2521 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     2581 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     2641 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     2701 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     2761 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     2821 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     2881 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2941 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3001 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     3061 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3121 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3181 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3241 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3301 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3361 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3421 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3481 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3541 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     3601 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     3661 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     3721 cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     3781 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     3841 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     3901 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     3961 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     4021 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     4081 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     4141 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
     4201 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg
     4261 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt
     4321 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
     4381 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt
     4441 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat
     4501 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc
     4561 acgatcatgc gcacccgtgg ccaggaccca acgctgcccg agatgcgccg cgtgcggctg
     4621 ctggagatgg cggacgcgat ggatatgttc tgccaagggt tggtttgcgc attcacagtt
     4681 ctccgcaaga attgattggc tccaattctt ggagtggtga atccgttagc gaggtgccgc
     4741 cggcttccat tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac gcggggaggc
     4801 agacaaggta tagggcggcg cctacaatcc atgccaaccc gttccatgtg ctcgccgagg
     4861 cggcataaat cgccgtgacg atcagcggtc caatgatcga agttaggctg gtaagagccg
     4921 cgagcgatcc ttgaagctgt ccctgatggt cgtcatctac ctgcctggac agcatggcct
     4981 gcaacgcggg catcccgatg ccgccggaag cgagaagaat cataatgggg aaggccatcc
     5041 agcctcgcgt cgcgaacgcc agcaagacgt agcccagcgc gtcggccgcc atgccggcga
     5101 taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga
     5161 gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa
     5221 agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga
     5281 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc
     5341 tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct
     5401 aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5461 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg
     5521 gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc
     5581 tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg
     5641 atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc
     5701 gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc
     5761 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg
     5821 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt
     5881 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt
     5941 aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc
     6001 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca
     6061 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca
     6121 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc
     6181 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt
     6241 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg
     6301 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg
     6361 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg
     6421 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg
     6481 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc
     6541 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc
     6601 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
     6661 cgccgccgca aggaatggtg catg
//
