LOCUS       sequence-161349-        7065 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Nuf2 N-terminally labeled with mTurquoise2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7065)
  AUTHORS   Yoo TY, Choi JM, Conway W, Yu CH, Pappu RV, Needleman DJ
  TITLE     Measuring NDC80 binding reveals the molecular basis of 
            tension-dependent kinetochore-microtubule attachments.
  JOURNAL   Elife. 2018 Jul 25;7. pii: 36392. doi: 10.7554/eLife.36392.
  PUBMED    30044223
REFERENCE   2  (bases 1 to 7065)
  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: "Elife."; 
            date: "2018-07-25"; volume: "7. pii: 36392. doi"; pages: " 
            10.7554/eLife.36392"
COMMENT     Sequence Label: pJAG98-mTurquoise2-Nuf2
FEATURES             Location/Qualifiers
     source          1..7065
                     /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             934..1350
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1318..1334
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1358..1367
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1364..1366,1367..1369,1370..2080)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1409..1430)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1670..1689)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2017..2038
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3559..3579)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3564..3893
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3744..3879
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3901..4299
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4346..4939
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5140..5157)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5311..5899)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5391..5410)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6077..6868,6869..6937))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6700..6719
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6938..7042)
                     /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 taagctggcc 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 tgctgcagca tcgttctgtg ttgtctctgt
      901 ctgactgtgt ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt
      961 ttgaccttag atcactggaa agatgtcgag cggctcgctc acaaccagtc ggtagatgtc
     1021 aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg
     1081 ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca
     1141 cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct
     1201 tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttctt
     1261 ccatccgcgc cgtctctccc ccttgaacct cctctttcga ccccgcctca atcctccctt
     1321 tatccagccc tcactccttc tctaggcgcc ggccgtcgcc accatggtga gcaagggcga
     1381 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     1441 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     1501 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgtc
     1561 ctggggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
     1621 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     1681 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     1741 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     1801 ctttagcgac aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt
     1861 caagatccgc cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa
     1921 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     1981 caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     2041 cgccgccggg atcactctcg gcatggacga gctgtacaag tactcagatc tgatggaaac
     2101 tttgtctttc cccagatata atgtagctga gattgtgatt catattcgca ataagatctt
     2161 aacaggagct gatggtaaaa acctcaccaa gaatgatctt tatccaaatc caaagcctga
     2221 agtcttgcac atgatctaca tgagagcctt acaaatagtc tatggaattc gactggaaca
     2281 tttttacatg atgccagtga actctgaagt catgtatcca catttaatgg aaggcttctt
     2341 accattcagc aatttagtta ctcatctgga ctcatttttg cctatctgcc gggtgaatga
     2401 ctttgagact gctgatattc tatgtccaaa agcaaaacgg acaagtcggt ttttaagtgg
     2461 cattatcaac tttattcact tcagagaagc atgccgtgaa acgtatatgg aatttctttg
     2521 gcaatataaa tcctctgcgg acaaaatgca acagttaaac gccgcacacc aggaggcatt
     2581 aatgaaactg gagagacttg attctgttcc agttgaagag caagaagagt tcaagcagct
     2641 ttcagatgga attcaggagc tacaacaatc actaaatcag gattttcatc aaaaaacgat
     2701 agtgctgcaa gagggaaatt cccaaaagaa gtcaaatatt tcagagaaaa ccaagcgttt
     2761 gaatgaacta aaattgttgg tggtttcttt gaaagaaata caagagagtt tgaaaacaaa
     2821 aattgtggat tctccagaga agttaaagaa ttataaagaa aaaatgaaag atacggtcca
     2881 gaagcttaaa aatgccagac aagaagtggt ggagaaatat gaaatctatg gagactcagt
     2941 tgactgcctg ccttcatgtc agttggaagt gcagttatat caaaagaaaa tacaggacct
     3001 ttcagataat agggaaaaat tagccagtat cttaaaggag agcctgaact tggaggacca
     3061 aattgagagt gatgagtcag aactgaagaa attgaagact gaagaaaatt cgttcaaaag
     3121 actgatgatt gtgaagaagg aaaaacttgc cacagcacaa ttcaaaataa ataagaagca
     3181 tgaagatgtt aagcaataca aacgcacagt aattgaggat tgcaataaag ttcaagaaaa
     3241 aagaggtgct gtctatgaac gagtaaccac aattaatcaa gaaatccaaa aaattaaact
     3301 tggaattcaa caactaaaag atgctgctga aagggagaaa ctgaagtccc aggaaatatt
     3361 tctaaacttg aaaactgctt tggagaaata ccacgacggt attgaaaagg cagcagagga
     3421 ctcctatgct aagatagatg agaagacagc tgaactgaag aggaagatgt tcaaaatgtc
     3481 aaccggatcc accggatcta gataactgat cagaattctg cagtcgacgg tacccgccag
     3541 cacagtggtc gaccctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3601 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3661 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3721 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3781 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     3841 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagcttc
     3901 atggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac ggctacaatc
     3961 aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc tagcgacggc
     4021 cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc agaactcgtg
     4081 gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga
     4141 aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct tctcgatctg
     4201 catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc agttgggatt
     4261 cgtgaattgc tgccctctgg ttatgtgtgg gagggctaag atcgataaaa taaaagattt
     4321 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
     4381 cttaagtaac gccattttgc aaggcatgga aaaatacata actgagaata gagaagttca
     4441 gatcaaggtc aggaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa
     4501 gcagttcctg ccccggctca gggccaagaa cagatggaac agctgaatat gggccaaaca
     4561 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga
     4621 tgcggtccag ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg
     4681 acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt
     4741 cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact cggggcgcca
     4801 gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg
     4861 catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg
     4921 tcagcggggg tctttcattt ccgacttgtg gtctcgctgc cttgggaggg tctcctctga
     4981 gtgattgact acccgtcagc gggggtcttc acatgcagca tgtatcaaaa ttaatttggt
     5041 tttttttctt aagtatttac attaaatggc catagttgca ttaatgaatc ggccaacgcg
     5101 cggggagagg cggtttgcgt attggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5161 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5221 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5281 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5341 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5401 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5461 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5521 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5581 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5641 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5701 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     5761 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5821 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5881 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5941 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6001 gatcctttta aattaaaaat gaagtttgcg gccgcaaatc aatctaaagt atatatgagt
     6061 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6121 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6181 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6241 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6301 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6361 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6421 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6481 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6541 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6601 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6661 tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca
     6721 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6781 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6841 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6901 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6961 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7021 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccac
//
