LOCUS       sequence-173661-        8031 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the C-terminal fragment of BRCA1 that is endogenously 
            generated by caspase cleavage in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8031)
  AUTHORS   Xu Z, Payoe R, Fahlman RP
  TITLE     The C-terminal proteolytic fragment of the breast cancer 
            susceptibility type 1 protein (BRCA1) is degraded by the N-end rule 
            pathway.
  JOURNAL   J Biol Chem. 2012 Mar 2;287(10):7495-502. doi: 
            10.1074/jbc.M111.301002. Epub 2012 Jan 18.
  PUBMED    22262859
REFERENCE   2  (bases 1 to 8031)
  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: 
            "10.1074/jbc.M111.301002"; journalName: "J Biol Chem"; date: 
            "2012-03-2- 2"; volume: "287"; issue: "10"; pages: "7495-502"
COMMENT     Sequence Label: RF-118-UB-BRCA1
FEATURES             Location/Qualifiers
     source          1..8031
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             920..1147
                     /codon_start=1
                     /product="human ubiquitin"
                     /label=ubiquitin
                     /translation="MEIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             3356..3421
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3456..3473)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3462..3686
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3732..4160
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3819..3838)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4029..4050
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4169..4189)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4174..4503
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4354..4489
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4416..4435
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4552..5577
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4552..5577
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    5707..5828
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5744..5763)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5798..5817
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5877..5893)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5877..5893)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5890..5912)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5901..5917
                     /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)."
     promoter        complement(join(5925..5931,5932..5949,5950..5955))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5970..5991
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6108..6125)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6279..6864)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6359..6378)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7035..7826,7827..7895))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7658..7677
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7896..8000)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttgcca tggagatctt cgtgaagacc ctgaccggca agaccatcac
      961 tctggaggtg gagcccagtg acaccatcga aaatgtgaaa gccaagatcc aagataaaga
     1021 aggcatcccc cccgaccagc agaggctcat ctttgcaggc aagcagctgg aagatggccg
     1081 cactctttcc gactacaaca tccagaaaga gtcgaccctg cacctggtcc tgcgcctgag
     1141 gggtggagat ggtgaaataa aggaagatac tagttttgct gaaaatgaca ttaaggaaag
     1201 ttctgctgtt tttagcaaaa gcgtccagag aggagagctt agcaggagtc ctagcccttt
     1261 cacccataca catttggctc agggttaccg aagaggggcc aagaaattag agtcctcaga
     1321 agagaactta tctagtgagg atgaagagct tccctgcttc caacacttgt tatttggtaa
     1381 agtaaacaat ataccttctc agtctactag gcatagcacc gttgctaccg agtgtctgtc
     1441 taagaacaca gaggagaatt tattatcatt gaagaatagc ttaaatgact gcagtaacca
     1501 ggtaatattg gcaaaggcat ctcaggaaca tcaccttagt gaggaaacaa aatgttctgc
     1561 tagcttgttt tcttcacagt gcagtgaatt ggaagacttg actgcaaata caaacaccca
     1621 ggatcctttc ttgattggtt cttccaaaca aatgaggcat cagtctgaaa gccagggagt
     1681 tggtctgagt gacaaggaat tggtttcaga tgatgaagaa agaggaacgg gcttggaaga
     1741 aaataatcaa gaagagcaaa gcatggattc aaacttaggt gaagcagcat ctgggtgtga
     1801 gagtgaaaca agcgtctctg aagactgctc agggctatcc tctcagagtg acattttaac
     1861 cactcagcag agggatacca tgcaacataa cctgataaag ctccagcagg aaatggctga
     1921 actagaagct gtgttagaac agcatgggag ccagccttct aacagctacc cttccatcat
     1981 aagtgactcc tctgcccttg aggacctgcg aaatccagaa caaagcacat cagaaaaaga
     2041 ttcgcatata catggccaaa gggacaactc catgttttct aaaaggccta gagaacatat
     2101 atcagtatta acttcacaga aaagtagtga ataccctata agccagaatc cagaaggcct
     2161 ttctgctgac aagtttgagg tgtctgcaga tagttctacc agtaaaaata aagaaccagg
     2221 agtggaaagg tcatcccctt ctaaatgccc atcattagat gataggtggt acatgcacag
     2281 ttgctctggg agtcttcaga atagaaacta cccatctcaa gaggggctca ttaaggttgt
     2341 tgatgtggag gagcaacagc tggaagagtc tgggccacac gatttgacgg aaacatctta
     2401 cttgccaagg caagatctag agggaacccc ttacctggaa tctggaatca gcctcttctc
     2461 tgatgaccct gaatctgatc cttctgaaga cagagcccca gagtcagctc gtgttggcaa
     2521 cataccatct tcaacctctg cattgaaagt tccccaattg aaagttgcag aatctgccca
     2581 gggtccagct gctgctcata ctactgatac tgctgggtat aatgcaatgg aagaaagtgt
     2641 gagcagggag aagccagaat tgacagcttc aacagaaagg gtcaacaaaa gaatgtccat
     2701 agtggtgtct ggcctgaccc cagaagaatt tatgctcgtg tacaagtttg ccagaaaaca
     2761 ccacatcact ttaactaatc taattactga agagactact catgttgtta tgaaaacaga
     2821 tgctgagttt gtgtgtgaac ggacactgaa atattttcta ggaattgcgg gaggaaaatg
     2881 ggtagttagc tatttctggg tgacccagtc tattaaagaa agaaaaatgc tgaatgagca
     2941 tgattttgaa gtcagaggag atgtggtcaa tggaagaaac caccaaggtc caaagcgagc
     3001 aagagaatcc caggacagaa agatcttcag ggggctagaa atctgttgct atgggccctt
     3061 caccaacatg cccacagatc aactggaatg gatggtacag ctgtgtggtg cttctgtggt
     3121 gaaggagctt tcatcattca cccttggcac aggtgtccac ccaattgtgg ttgtgcagcc
     3181 agatgcctgg acagaggaca atggcttcca tgcaattggg cagatgtgtg aggcacctgt
     3241 ggtgacccga gagtgggtgt tggacagtgt agcactctac cagtgccagg agctggacac
     3301 ctacctgata ccccagatcc cccacagcca ctacctcgag tctagaatgg ctagcgacta
     3361 caaagaccat gacggtgatt ataaagatca tgacatcgat tacaaggatg acgatgacaa
     3421 gtgatctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     3481 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     3541 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3601 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     3661 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     3721 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3781 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3841 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3901 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3961 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4021 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4081 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4141 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4201 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4261 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4321 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4381 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4441 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4501 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt gatgaaaaag
     4561 cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc
     4621 gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg
     4681 cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga tcgttatgtt
     4741 tatcggcact ttgcatcggc cgcgctcccg attccggaag tgcttgacat tggggaattc
     4801 agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg
     4861 cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct
     4921 gcggccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa
     4981 tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta tcactggcaa
     5041 actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt
     5101 tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg ctccaacaat
     5161 gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc gatgttcggg
     5221 gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc ttgtatggag
     5281 cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc gcggctccgg
     5341 gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga cggcaatttc
     5401 gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg agccgggact
     5461 gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa
     5521 gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa ggaatagcac
     5581 gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     5641 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5701 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5761 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5821 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     5881 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     5941 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     6001 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     6061 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     6121 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     6181 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6241 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6301 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6361 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6421 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6481 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6541 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6601 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6661 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     6721 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6781 tcttgatccg gcaaacaaac caccgctggt agcggttttt ttgtttgcaa gcagcagatt
     6841 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6901 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6961 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     7021 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     7081 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     7141 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     7201 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     7261 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     7321 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     7381 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     7441 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7501 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     7561 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     7621 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     7681 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7741 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7801 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7861 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7921 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7981 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
