LOCUS       sequence-343357-        7320 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stable expression of fluorescent Separase cleavage sensor with human
            Rec8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7320)
  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 7320)
  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)-mNeonGreen
FEATURES             Location/Qualifiers
     source          1..7320
                     /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      3438..3447
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3905..3925)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3910..4239
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4090..4225
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4247..4645
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4814..5283
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5413..5430)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5584..6172)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5664..5683)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6332..7123,7124..7192))
                     /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     6955..6974
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7193..7297)
                     /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 cggaccggta ggcggcggtg
     3181 ggtccgtgag caagggcgag gaggacaaca tggccagcct gcccgccacc cacgagctgc
     3241 acatcttcgg cagcatcaac ggcgtggact tcgacatggt gggccagggc accggcaacc
     3301 ccaacgacgg ctacgaggag ctgaacctga agagcaccaa gggcgacctg cagttcagcc
     3361 cctggattct ggtgccccac atcggctacg gcttccacca gtacctgccc taccccgacg
     3421 gcatgagccc cttccaggcc gccatggtgg acggcagcgg ctaccaggtg caccgcacca
     3481 tgcagttcga ggacggcgcc agcctgaccg tgaactaccg ctacacctac gagggcagcc
     3541 acatcaaggg cgaggcccag gtgaagggca ccggcttccc cgccgacggc cccgtgatga
     3601 ccaacagcct gaccgccgcc gactggtgcc gcagcaagaa gacctacccc aacgacaaga
     3661 ccatcatcag caccttcaag tggagctaca ccaccggcaa cggcaagcgc taccgcagca
     3721 ccgcccgcac cacctacacc ttcgccaagc ccatggccgc caactacctg aagaaccagc
     3781 ccatgtacgt gttccgcaag accgagctga agcacagcaa gaccgagctg aacttcaagg
     3841 agtggcagaa ggccttcacc gacgtgatgg gcatggacga gctgtacaag taagtcgacc
     3901 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3961 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4021 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4081 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4141 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     4201 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttcatgg ccaagccttt
     4261 gtctcaagaa gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat
     4321 ctctgaagac tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg
     4381 tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc
     4441 tgctgctgcg gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg
     4501 catcttgagc ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa
     4561 agccatagtg aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc
     4621 ctctggttat gtgtgggagg gctaagatcg ataaaataaa agattttatt tagtctccag
     4681 aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca
     4741 ttttgcaagg catggaaaaa tacataactg agaatagaga agttcagatc aaggtcagga
     4801 acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
     4861 ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat atctgtggta
     4921 agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg gtccagccct
     4981 cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc
     5041 tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct
     5101 ccccgagctc aataaaagag cccacaaccc ctcactcggg gcgccagtcc tccgattgac
     5161 tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc cgacttgtgg
     5221 tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag cgggggtctt
     5281 tcacatgcag catgtatcaa aattaatttg gttttttttc ttaagtattt acattaaatg
     5341 gccatagttg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5401 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5461 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5521 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5581 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5641 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5701 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5761 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5821 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5881 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5941 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6001 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6061 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6121 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6181 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6241 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttgcggccgc aaatcaatct
     6301 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6361 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6421 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6481 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6541 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6601 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6661 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6721 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6781 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6841 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6901 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6961 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7021 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7081 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7141 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7201 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7261 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
//
