LOCUS       sequence-343356-        7656 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stable expression of human Meikin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7656)
  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 7656)
  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: 2xmNeonGreen-hMeikin
FEATURES             Location/Qualifiers
     source          1..7656
                     /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"
     regulatory      1369..1378
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1630..1639
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2344..2353
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2842..2862
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2875..2919
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(4241..4261)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4246..4575
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4426..4561
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4583..4981
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             5150..5619
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5749..5766)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5920..6508)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6000..6019)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6668..7459,7460..7528))
                     /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     7291..7310
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7529..7633)
                     /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 ccggatccgc caccatggtg
     1381 agcaagggcg aggaggacaa catggccagc ctgcccgcca cccacgagct gcacatcttc
     1441 ggcagcatca acggcgtgga cttcgacatg gtgggccagg gcaccggcaa ccccaacgac
     1501 ggctacgagg agctgaacct gaagagcacc aagggcgacc tgcagttcag cccctggatt
     1561 ctggtgcccc acatcggcta cggcttccac cagtacctgc cctaccccga cggcatgagc
     1621 cccttccagg ccgccatggt ggacggcagc ggctaccagg tgcaccgcac catgcagttc
     1681 gaggacggcg ccagcctgac cgtgaactac cgctacacct acgagggcag ccacatcaag
     1741 ggcgaggccc aggtgaaggg caccggcttc cccgccgacg gccccgtgat gaccaacagc
     1801 ctgaccgccg ccgactggtg ccgcagcaag aagacctacc ccaacgacaa gaccatcatc
     1861 agcaccttca agtggagcta caccaccggc aacggcaagc gctaccgcag caccgcccgc
     1921 accacctaca ccttcgccaa gcccatggcc gccaactacc tgaagaacca gcccatgtac
     1981 gtgttccgca agaccgagct gaagcacagc aagaccgagc tgaacttcaa ggagtggcag
     2041 aaggccttca ccgacgtgat gggcatggac gagctgtaca agtccggaat ggtgagcaag
     2101 ggcgaggagg acaacatggc cagcctgccc gccacccacg agctgcacat cttcggcagc
     2161 atcaacggcg tggacttcga catggtgggc cagggcaccg gcaaccccaa cgacggctac
     2221 gaggagctga acctgaagag caccaagggc gacctgcagt tcagcccctg gattctggtg
     2281 ccccacatcg gctacggctt ccaccagtac ctgccctacc ccgacggcat gagccccttc
     2341 caggccgcca tggtggacgg cagcggctac caggtgcacc gcaccatgca gttcgaggac
     2401 ggcgccagcc tgaccgtgaa ctaccgctac acctacgagg gcagccacat caagggcgag
     2461 gcccaggtga agggcaccgg cttccccgcc gacggccccg tgatgaccaa cagcctgacc
     2521 gccgccgact ggtgccgcag caagaagacc taccccaacg acaagaccat catcagcacc
     2581 ttcaagtgga gctacaccac cggcaacggc aagcgctacc gcagcaccgc ccgcaccacc
     2641 tacaccttcg ccaagcccat ggccgccaac tacctgaaga accagcccat gtacgtgttc
     2701 cgcaagaccg agctgaagca cagcaagacc gagctgaact tcaaggagtg gcagaaggcc
     2761 ttcaccgacg tgatgggcat ggacgagctg tacaagaccg gaggggatcg gtggagctcc
     2821 accggtggcg gccgctctag agagaatctt tattttcagg gcgccgccaa attcaaagaa
     2881 accgctgctg ctaaattcga acgccagcac atggacagcg gaggtggagg tactagtctc
     2941 gagatgtggc ccctgagggt gtacaccaga aagaagaggg agggccagag actgaacctg
     3001 acacccacac ctgacctggg ctcccctgcc aaggctgaag ctcctcccgg cagcaagagg
     3061 aaaggcaagg tgcacggcct gagcaagatc gctgagaaag ccgagagaag caggcagggc
     3121 ggatccggaa gcggaccttt cagccccagg ctcggagtga ccggcgagaa gtccctgcag
     3181 gagaacaggt ccagcgagga cacccaggac gagaagattg cttccctgag ggagagcgtg
     3241 accgacgacc tgcaggtgga ctccagcagc tccaactccg aactggtgtc cggactgagc
     3301 ctgcatcacg gcatggcctc ctccctgctg tcctatagcg tgaccgattc ctacgccgag
     3361 tacaagagct tcgaggagtc cttcccctcc cctgagctgt tcaggaagtc cgactacctg
     3421 gactgggagt gccccaatct ggaggagcac atgcagtgga agaacagcac cctgctcgac
     3481 acctccaagg ccgtggctat cgagaaggct ccccagttca gcaacgtcag cgccatcttc
     3541 tccaccagca gcgaggacta ccagaagtgc cacaggaaga ccgtgatgac cgtggccgat
     3601 cagaacgtgt cccccaaggc caagtgcgcc agcaatagcg agagcgacaa cgctgcctgt
     3661 gaaatcctgc tggccgagaa gacatgcccc agcacccccg agaagaccaa gaagaagaag
     3721 accaacagca gcacacctgg caagaagaac agaggactgc tgacctccac acctagctcc
     3781 gagaccgccg gctttgtgat cgacctgagc tccgtgcaga aggcctcctt tgaggagctg
     3841 ttccccaacg tgagcaacta cgtgaacagc aacgagatcg tccccgtgtc cagcctccag
     3901 gagaacagct ccaacgagtt ccccgccaac gcctccgaga tctgctgcat catcaggacc
     3961 tcccctggca ccaggcaggt gaaaaacaag ggcgtgattg tgaagaagaa aaagtacagc
     4021 ctgcccaagg acacccccca ggacatcatc atcaagatgg cctaagaatt ctgcagtcga
     4081 cggtaccgcg ggcccgggat ccaccggatc tagataactg atcataatca gccataccac
     4141 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     4201 taaaatgaat gcaattcgcc agcacagtgg tcgaccctgt ggaatgtgtg tcagttaggg
     4261 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4321 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4381 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4441 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4501 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4561 ctaggctttt gcaaaaagct tcatggccaa gcctttgtct caagaagaat ccaccctcat
     4621 tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca gcgtcgccag
     4681 cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc attttactgg
     4741 gggaccttgt gcagaactcg tggtgctggg cactgctgct gctgcggcag ctggcaacct
     4801 gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct gcggacggtg
     4861 ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg
     4921 acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt gggagggcta
     4981 agatcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc
     5041 acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaaataca
     5101 taactgagaa tagagaagtt cagatcaagg tcaggaacag atggaacagc tgaatatggg
     5161 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatgga
     5221 acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
     5281 ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga gaaccatcag
     5341 atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat
     5401 cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca
     5461 caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt acccgtgtat
     5521 ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct
     5581 cctctgagtg attgactacc cgtcagcggg ggtctttcac atgcagcatg tatcaaaatt
     5641 aatttggttt tttttcttaa gtatttacat taaatggcca tagttgcatt aatgaatcgg
     5701 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5761 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5821 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5881 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5941 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6001 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6061 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     6121 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6181 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6241 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6301 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6361 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6421 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6481 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6541 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6601 cttcacctag atccttttgc ggccgcaaat caatctaaag tatatatgag taaacttggt
     6661 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6721 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6781 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6841 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6901 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6961 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7021 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7081 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7141 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7201 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7261 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7321 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7381 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7441 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7501 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7561 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7621 taggggttcc gcgcacattt ccccgaaaag tgccac
//
