LOCUS       sequence-173660-        7140 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the C-terminal fragment of BMX 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 7140)
  TITLE     Fahlman lab unpublished plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7140)
  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            "Unpublished"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: RF-227-UB-BMX
FEATURES             Location/Qualifiers
     source          1..7140
                     /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             2468..2533
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2568..2585)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2574..2798
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2844..3272
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2931..2950)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3141..3162
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3281..3301)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3286..3615
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3466..3601
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3528..3547
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3664..4689
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3664..4689
                     /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    4819..4940
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4856..4875)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4910..4929
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4989..5005)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4989..5005)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5002..5024)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5013..5029
                     /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(5037..5043,5044..5061,5062..5067))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5082..5103
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5220..5237)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5391..5976)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5471..5490)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6147..6938,6939..7007))
                     /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     6770..6789
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7008..7112)
                     /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 gggtggatgg tggcaagtaa gaaaactgaa aagtagcagc agcagtgaag atgttgcaag
     1201 cagtaaccaa aaagaaagaa atgtgaatca caccacctca aagatttcat gggaattccc
     1261 tgagtcaagt tcatctgaag aagaggaaaa cctggatgat tatgactggt ttgctggtaa
     1321 catctccaga tcacaatctg aacagttact cagacaaaag ggaaaagaag gagcatttat
     1381 ggttagaaat tcgagccaag tgggaatgta cacagtgtcc ttatttagta aggctgtgaa
     1441 tgataaaaaa ggaactgtca aacattacca cgtgcataca aatgctgaga acaaattata
     1501 cctggcagaa aactactgtt ttgattccat tccaaagctt attcattatc atcaacacaa
     1561 ttcagcaggc atgatcacac ggctccgcca ccctgtgtca acaaaggcca acaaggtccc
     1621 cgactctgtg tccctgggaa atggaatctg ggaactgaaa agagaagaga ttaccttgtt
     1681 gaaggagctg ggaagtggcc agtttggagt ggtccagctg ggcaagtgga aggggcagta
     1741 tgatgttgct gttaagatga tcaaggaggg ctccatgtca gaagatgaat tctttcagga
     1801 ggcccagact atgatgaaac tcagccatcc caagctggtt aaattctatg gagtgtgttc
     1861 aaaggaatac cccatataca tagtgactga atatataagc aatggctgct tgctgaatta
     1921 cctgaggagt cacggaaaag gacttgaacc ttcccagctc ttagaaatgt gctacgatgt
     1981 ctgtgaaggc atggccttct tggagagtca ccaattcata caccgggact tggctgctcg
     2041 taactgcttg gtggacagag atctctgtgt gaaagtatct gactttggaa tgacaaggta
     2101 tgttcttgat gaccagtatg tcagttcagt cggaacaaag tttccagtca agtggtcagc
     2161 tccagaggtg tttcattact tcaaatacag cagcaagtca gacgtatggg catttgccat
     2221 cctgatgtgg gaggtgttca gcctggggaa gcagccctat gacttgtatg acaactccca
     2281 ggtggttctg aaggtctccc agggccacag gctttaccgg ccccacctgg catcggacac
     2341 catctaccag atcatgtaca gctgctggca cgagcttcca gaaaagcgtc ccacatttca
     2401 gcaactcctg tcttccattg aaccacttcg ggaaaaagac aagcatctcg agtctagaat
     2461 ggctagcgac tacaaagacc atgacggtga ttataaagat catgacatcg attacaagga
     2521 tgacgatgac aagtgatcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     2581 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     2641 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     2701 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     2761 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     2821 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2881 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     2941 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3001 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3061 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3121 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3181 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3241 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     3301 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3361 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3421 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3481 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3541 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3601 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac
     3661 gtgatgaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc
     3721 gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc
     3781 gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa
     3841 gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac
     3901 attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg
     3961 ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggccatg
     4021 gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa
     4081 ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg
     4141 tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat
     4201 gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc
     4261 ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag
     4321 gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg
     4381 gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg
     4441 ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt
     4501 gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc
     4561 ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat
     4621 ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca
     4681 aaggaatagc acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4741 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4801 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4861 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4921 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     4981 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5041 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     5101 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5161 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5221 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5281 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5341 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5401 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5461 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5521 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5581 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5641 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5701 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5761 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5821 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5881 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc
     5941 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6001 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6061 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6121 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6181 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6241 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6301 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6361 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6421 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6481 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6541 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6601 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6661 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6721 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6781 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6841 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6901 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6961 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7021 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7081 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
//
