LOCUS       sequence-416909-        8002 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses BubR1-D911A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8002)
  TITLE     Expresses TFAM in mammalian cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8002)
  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            "Unpublished"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: pCAGGS-FLAG-BubR1-D911A
FEATURES             Location/Qualifiers
     source          1..8002
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1739..1762
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(5023..5042)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5088..5143
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5142..5161)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5504..5520)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5504..5520)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5517..5539)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5528..5544
                     /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(5552..5558,5559..5576,5577..5582))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5597..5618
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5677..5872
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5723..5858
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5878..6012
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5915..5934)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5969..5988
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6080..6097)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6251..6839)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6331..6350)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7010..7801,7802..7870))
                     /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     7633..7652
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7871..7975)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg gggggggggc gcccgggggg gggggggggg gggggggggg
      541 gggggggggg ggggggggcg gggggggggg gggcgcgcca gggggggggg gggggcggag
      601 gccttttttt gcgggggggg gggggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcaccatgga
     1741 ttacaaggat gacgacgata agtgcggccg cgcggcggtg aagaaggaag ggggtgctct
     1801 gagtgaagcc atgtccctgg agggagatga atgggaactg agtaaagaaa atgtacaacc
     1861 tttaaggcaa gggcggatca tgtccacgct tcagggagca ctggcacaag aatctgcctg
     1921 taacaatact cttcagcagc agaaacgggc atttgaatat gaaattcgat tttacactgg
     1981 aaatgaccct ctggatgttt gggataggta tatcagctgg acagagcaga actatcctca
     2041 aggtgggaag gagagtaata tgtcaacgtt attagaaaga gctgtagaag cactacaagg
     2101 agaaaaacga tattatagtg atcctcgatt tctcaatctc tggcttaaat tagggcgttt
     2161 atgcaatgag cctttggata tgtacagtta cttgcacaac caagggattg gtgtttcact
     2221 tgctcagttc tatatctcat gggcagaaga atatgaagct agagaaaact ttaggaaagc
     2281 agatgcgata tttcaggaag ggattcaaca gaaggctgaa ccactagaaa gactacagtc
     2341 ccagcaccga caattccaag ctcgagtgtc tcggcaaact ctgttggcac ttgagaaaga
     2401 agaagaggag gaagtttttg agtcttctgt accacaacga agcacactag ctgaactaaa
     2461 gagcaaaggg aaaaagacag caagagctcc aatcatccgt gtaggaggtg ctctcaaggc
     2521 tccaagccag aacagaggac tccaaaatcc atttcctcaa cagatgcaaa ataatagtag
     2581 aattactgtt tttgatgaaa atgctgatga ggcttctaca gcagagttgt ctaagcctac
     2641 agtccagcca tggatagcac cccccatgcc cagggccaaa gagaatgagc tgcaagcagg
     2701 cccttggaac acaggcaggt ccttggaaca caggcctcgt ggcaatacag cttcactgat
     2761 agctgtaccc gctgtgcttc ccagtttcac tccatatgtg gaagagactg cacaacagcc
     2821 agttatgaca ccatgtaaaa ttgaacctag tataaaccac atcctaagca ccagaaagcc
     2881 tggaaaggaa gaaggagatc ctctacaaag ggttcagagc catcagcaag catctgagga
     2941 gaagaaagag aagatgatgt attgtaagga gaagatttat gcaggagtag gggaattctc
     3001 ctttgaagaa attcgggctg aagttttccg gaagaaatta aaagagcaaa gggaagccga
     3061 gctattgacc agtgcagaga agagagcaga aatgcagaaa cagattgaag agatggagaa
     3121 gaagctaaaa gaaatccaaa ctactcagca agaaagaaca ggtgatcagc aagaagagac
     3181 gatgcctaca aaggagacaa ctaaactgca aattgcttcc gagtctcaga aaataccagg
     3241 aatgactcta tccagttctg tttgtcaagt aaactgttgt gccagagaaa cttcacttgc
     3301 ggagaacatt tggcaggaac aacctcattc taaaggtccc agtgtacctt tctccatttt
     3361 tgatgagttt cttctttcag aaaagaagaa taaaagtcct cctgcagatc ccccacgagt
     3421 tttagctcaa cgaagacccc ttgcagttct caaaacctca gaaagcatca cctcaaatga
     3481 agatgtgtct ccagatgttt gtgatgaatt tacaggaatt gaacccttga gcgaggatgc
     3541 cattatcaca ggcttcagaa atgtaacaat ttgtcctaac ccagaagaca cttgtgactt
     3601 tgccagagca gctcgttttg tatccactcc ttttcatgag ataatgtcct tgaaggatct
     3661 cccttctgat cctgagagac tgttaccgga agaagatcta gatgtaaaga cctctgagga
     3721 ccagcagaca gcttgtggca ctatctacag tcagactctc agcatcaaga agctgagccc
     3781 aattattgaa gacagtcgtg aagccacaca ctcctctggc ttctctggtt cttctgcctc
     3841 ggttgcaagc acctcctcca tcaaatgtct tcaaattcct gagaaactag aacttactaa
     3901 tgagacttca gaaaacccta ctcagtcacc atggtgttca cagtatcgca gacagctact
     3961 gaagtcccta ccagagttaa gtgcctctgc agagttgtgt atagaagaca gaccaatgcc
     4021 taagttggaa attgagaagg aaattgaatt aggtaatgag gattactgca ttaaacgaga
     4081 atacctaata tgtgaagatt acaagttatt ctgggtggcg ccaagaaact ctgcagaatt
     4141 aacagtaata aaggtatctt ctcaacctgt cccatgggac ttttatatca acctcaagtt
     4201 aaaggaacgt ttaaatgaag attttgatca tttttgcagc tgttatcaat atcaagatgg
     4261 ctgtattgtt tggcaccaat atataaactg cttcaccctt caggatcttc tccaacacag
     4321 tgaatatatt acccatgaaa taacagtgtt gattatttat aaccttttga caatagtgga
     4381 gatgctacac aaagcagaaa tagtccatgg tgacttgagt ccaaggtgtc tgattctcag
     4441 aaacagaatc cacgatccct atgattgtaa caagaacaat caagctttga agatagtggc
     4501 attttcctac agtgttgacc ttagggtgca gctggatgtt tttaccctca gcggctttcg
     4561 gactgtacag atcctggaag gacaaaagat cctggctaac tgttcttctc cctaccaggt
     4621 agacctgttt ggtatagcag atttagcaca tttactattg ttcaaggaac acctacaggt
     4681 cttctgggat gggtccttct ggaaacttag ccaaaatatt tctgagctaa aagatggtga
     4741 attgtggaat aaattctttg tgcggattct gaatgccaat gatgaggcca cagtgtctgt
     4801 tcttggggag cttgcagcag aaatgaatgg ggtttttgac actacattcc aaagtcacct
     4861 gaacaaagcc ttatggaagg tagggaagtt aactagtcct ggggctttgc tctttcagtg
     4921 aacgcgtgaa ggtaccaggg aattcactcc tcaggtgcag gctgcctatc agaaggtggt
     4981 ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa
     5041 aaattatggg gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt
     5101 tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg
     5161 gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc
     5221 ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca
     5281 gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt
     5341 tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt
     5401 actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt
     5461 atggagatcc ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg
     5521 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5581 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5641 ctttccagtc gggaaacctg tcgtgccagc ggatccgcat ctcaattagt cagcaaccat
     5701 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     5761 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     5821 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctaac
     5881 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     5941 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     6001 catgtctgga tccgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     6061 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6121 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6181 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6241 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6301 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6361 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6421 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6481 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6541 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6601 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6661 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     6721 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6781 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6841 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6901 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6961 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7021 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7081 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7141 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7201 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7261 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7321 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7381 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7441 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7501 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7561 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7621 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7681 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7741 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7801 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7861 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7921 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7981 ttccccgaaa agtgccacct gg
//
