LOCUS       sequence-343358-        8370 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stable expression of fluorescent Separase cleavage sensor with human
            Rec8 fused to human Meikin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8370)
  AUTHORS   Maier NK, Ma J, Lampson MA, Cheeseman IM
  TITLE     Separase cleaves the kinetochore protein Meikin at the meiosis I/II 
            transition.
  JOURNAL   Dev Cell. 2021 Aug 9;56(15):2192-2206.e8. doi: 
            10.1016/j.devcel.2021.06.019. Epub 2021 Jul 30.
  PUBMED    34331869
REFERENCE   2  (bases 1 to 8370)
  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: 
            "10.1016/j.devcel.2021.06.019"; journalName: "Dev Cell"; date: 
            "2021-08-9- 9"; volume: "56"; issue: "15"; pages: "2192-2206.e8"
COMMENT     Sequence Label: H2B-mScarlet- hRec8(297-506)-hMeikin(1-332, E1...
FEATURES             Location/Qualifiers
     source          1..8370
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             8..477
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    542..741
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             942..1358
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1288..1310
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1326..1342
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1368..1745
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     CDS             1788..2480
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="VSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGGA
                     VTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKM
                     ALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHS
                     TGGMDELYK"
     primer_bind     complement(1918..1936)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2186..2205
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     regulatory      4488..4497
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4955..4975)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4960..5289
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5140..5275
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5297..5695
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             5864..6333
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6463..6480)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6634..7222)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6714..6733)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7382..8173,8174..8242))
                     /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     8005..8024
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8243..8347)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
       61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
      121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
      181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
      241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
      301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
      361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
      421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
      481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
      541 caagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
      601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
      661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
      721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
      781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
      841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt
      901 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
      961 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
     1021 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     1081 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     1141 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     1201 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     1261 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
     1321 cctcccttta tccagccctc actccttctc taggcgccgg cgccaccatg ccagagccag
     1381 cgaagtctgc tcccgccccg aaaaagggct ccaagaaggc ggtgactaag gcgcagaaga
     1441 aaggcggcaa gaagcgcaag cgcagccgca aggagagcta ttccatctat gtgtacaagg
     1501 ttctgaagca ggtccaccct gacaccggca tttcgtccaa ggccatgggc atcatgaatt
     1561 cgtttgtgaa cgacattttc gagcgcatcg caggtgaggc ttcccgcctg gcgcattaca
     1621 acaagcgctc gaccatcacc tccagggaga tccagacggc cgtgcgcctg ctgctgcctg
     1681 gggagttggc caagcacgcc gtgtccgagg gtactaaggc catcaccaag tacaccagcg
     1741 ctaagggtgg tggcggttca ggaagcgggt ccggctctgg gcgcgccgtg agcaagggcg
     1801 aggcagtgat caaggagttc atgcggttca aggtgcacat ggagggctcc atgaacggcc
     1861 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
     1921 agctgaaggt gaccaagggt ggccccctgc ccttctcctg ggacatcctg tcccctcagt
     1981 tcatgtacgg ctccagggcc ttcaccaagc accccgccga catccccgac tactataagc
     2041 agtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgccg
     2101 tgaccgtgac ccaggacacc tccctggagg acggcaccct gatctacaag gtgaagctcc
     2161 gcggcaccaa cttccctcct gacggccccg taatgcagaa gaagacaatg ggctgggaag
     2221 cgtccaccga gcggttgtac cccgaggacg gcgtgctgaa gggcgacatt aagatggccc
     2281 tgcgcctgaa ggacggcggc cgctacctgg cggacttcaa gaccacctac aaggccaaga
     2341 agcccgtgca gatgcccggc gcctacaacg tcgaccgcaa gttggacatc acctcccaca
     2401 acgaggacta caccgtggtg gaacagtacg aacgctccga gggccgacac tccaccggcg
     2461 gcatggacga gctgtacaag tccggagggg gctctggtgg aggcagcctc gagagatcta
     2521 cgcgtcctcg ccgccgccgt cgtcgccggt tactgttctg ggacaaggag actcagatct
     2581 ccccggagaa attccaggaa caactgcaaa ccagagccca ctgctgggaa tgtcctatgg
     2641 tgcagccgcc cgagaggacc atcagaggcc ctgcggagtt gttcagaacc ccaactctct
     2701 ctggctggct accccctgaa ctactgggtc tctggaccca ttgtgcccag ccacccccaa
     2761 aagccctcag gcgagagctg cctgaggagg cagccgctga ggaggaaagg agaaagattg
     2821 aagttccaag tgagattgag gtcccgaggg aggccctgga gcccagtgtt ccccttatgg
     2881 tgtctttaga gatctcccta gaggcagctg aagaggagaa gtcccgcatc agcctcatcc
     2941 caccagaaga acggtgggcc tggcctgagg tggaggcgcc agaagctcct gcattgcccg
     3001 tggtgcctga actccctgag gtgcccatgg agatgccttt ggtgctgccc ccagagctcg
     3061 agctgctctc actggaagca gtgcacaggg cagtggcact ggagctgcag gctaacaggg
     3121 agcccgactt cagcagcctg gtgtcacctc tcagcggatc tggtggtggc ggttcaggcg
     3181 gaggtggctc tatgtggccc ctgagggtgt acaccagaaa gaagagggag ggccagagac
     3241 tgaacctgac acccacacct gacctgggct cccctgccaa ggctgaagct cctcccggca
     3301 gcaagaggaa aggcaaggtg cacggcctga gcaagatcgc tgagaaagcc gagagaagca
     3361 ggcagggcgg atccggaagc ggacctttca gccccaggct cggagtgacc ggcgagaagt
     3421 ccctgcagga gaacaggtcc agcgaggaca cccaggacga gaagattgct tccctgaggg
     3481 agagcgtgac cgacgacctg caggtggact ccagcagctc caactccgaa ctggtgtccg
     3541 gactgagcct gcatcacggc atggcctcct ccctgctgtc ctatagcgtg accgattcct
     3601 acgccgagta caagagcttc gaggagtcct tcccctcccc tagactgttc gagaagtccg
     3661 actacctgga ctgggagtgc cccaatctgg aggagcacat gcagtggaag aacagcaccc
     3721 tgctcgacac ctccaaggcc gtggctatcg agaaggctcc ccagttcagc aacgtcagcg
     3781 ccatcttctc caccagcagc gaggactacc agaagtgcca caggaagacc gtgatgaccg
     3841 tggccgatca gaacgtgtcc cccaaggcca agtgcgccag caatagcgag agcgacaacg
     3901 ctgcctgtga aatcctgctg gccgagaaga catgccccag cacccccgag aagaccaaga
     3961 agaagaagac caacagcagc acacctggca agaagaacag aggactgctg acctccacac
     4021 ctagctccga gaccgccggc tttgtgatcg acctgagctc cgtgcagaag gcctcctttg
     4081 aggagctgtt ccccaacgtg agcaactacg tgaacagcaa cgagatcgtc cccgtgtcca
     4141 gcctccagga gaacagctcc aacgagttcc ccgccaacgc ctccgaggga agcggctccg
     4201 ggagtggatc cggaccggta ggcggcggtg ggtccgtgag caagggcgag gaggacaaca
     4261 tggccagcct gcccgccacc cacgagctgc acatcttcgg cagcatcaac ggcgtggact
     4321 tcgacatggt gggccagggc accggcaacc ccaacgacgg ctacgaggag ctgaacctga
     4381 agagcaccaa gggcgacctg cagttcagcc cctggattct ggtgccccac atcggctacg
     4441 gcttccacca gtacctgccc taccccgacg gcatgagccc cttccaggcc gccatggtgg
     4501 acggcagcgg ctaccaggtg caccgcacca tgcagttcga ggacggcgcc agcctgaccg
     4561 tgaactaccg ctacacctac gagggcagcc acatcaaggg cgaggcccag gtgaagggca
     4621 ccggcttccc cgccgacggc cccgtgatga ccaacagcct gaccgccgcc gactggtgcc
     4681 gcagcaagaa gacctacccc aacgacaaga ccatcatcag caccttcaag tggagctaca
     4741 ccaccggcaa cggcaagcgc taccgcagca ccgcccgcac cacctacacc ttcgccaagc
     4801 ccatggccgc caactacctg aagaaccagc ccatgtacgt gttccgcaag accgagctga
     4861 agcacagcaa gaccgagctg aacttcaagg agtggcagaa ggccttcacc gacgtgatgg
     4921 gcatggacga gctgtacaag taagtcgacc ctgtggaatg tgtgtcagtt agggtgtgga
     4981 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     5041 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     5101 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     5161 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     5221 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     5281 ttttgcaaaa agcttcatgg ccaagccttt gtctcaagaa gaatccaccc tcattgaaag
     5341 agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg ccagcgcagc
     5401 tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta ctgggggacc
     5461 ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca acctgacttg
     5521 tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac ggtgccgaca
     5581 ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg atggacagcc
     5641 gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg gctaagatcg
     5701 ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt
     5761 aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg
     5821 agaatagaga agttcagatc aaggtcagga acagatggaa cagctgaata tgggccaaac
     5881 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggaacagct
     5941 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
     6001 acagatggtc cccagatgcg gtccagccct cagcagtttc tagagaacca tcagatgttt
     6061 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
     6121 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
     6181 ctcactcggg gcgccagtcc tccgattgac tgagtcgccc gggtacccgt gtatccaata
     6241 aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg
     6301 agtgattgac tacccgtcag cgggggtctt tcacatgcag catgtatcaa aattaatttg
     6361 gttttttttc ttaagtattt acattaaatg gccatagttg cattaatgaa tcggccaacg
     6421 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6481 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6541 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6601 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6661 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6721 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6781 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6841 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6901 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6961 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7021 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     7081 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7141 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7201 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7261 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7321 ctagatcctt ttgcggccgc aaatcaatct aaagtatata tgagtaaact tggtctgaca
     7381 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     7441 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     7501 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7561 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7621 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7681 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7741 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7801 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7861 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7921 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7981 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     8041 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     8101 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     8161 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     8221 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     8281 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     8341 ttccgcgcac atttccccga aaagtgccac
//
