LOCUS       sequence-161095-        7320 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 7320)
  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 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"; journalName: "Elife."; 
            date: "2018-07-25"; volume: "7. pii: 36392. doi"; pages: " 
            10.7554/eLife.36392"
COMMENT     Sequence Label: pBABE-puro-mTurquoise2-Nuf2
FEATURES             Location/Qualifiers
     source          1..7320
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             7..476
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    541..740
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             933..1349
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1317..1333
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1357..1366
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1363..1365,1366..1368,1369..2079)
                     /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(1408..1429)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1669..1688)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2016..2037
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3558..3578)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3563..3892
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3743..3878
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3902..4501
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3902..3921)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4398..4418
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4601..5194
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5395..5412)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5566..6154)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5646..5665)
                     /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
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     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 tgcagcctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
       61 ggccaagaac agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt
      121 cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtccagc cctcagcagt
      181 ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct
      241 tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag
      301 ctcaataaaa gagcccacaa cccctcactc ggggcgccag tcctccgatt gactgagtcg
      361 cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct
      421 gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcatttg
      481 ggggctcgtc cgggatcggg agacccctgc ccagggacca ccgacccacc accgggaggt
      541 aagctggcca gcaacttatc tgtgtctgtc cgattgtcta gtgtctatga ctgattttat
      601 gcgcctgcgt cggtactagt tagctaacta gctctgtatc tggcggaccc gtggtggaac
      661 tgacgagttc tgaacacccg gccgcaaccc tgggagacgt cccagggact ttgggggccg
      721 tttttgtggc ccgacctgag gaagggagtc gatgtggaat ccgaccccgt caggatatgt
      781 ggttctggta ggagacgaga acctaaaaca gttcccgcct ccgtctgaat ttttgctttc
      841 ggtttggaac cgaagccgcg cgtcttgtct gctgcagcat cgttctgtgt tgtctctgtc
      901 tgactgtgtt tctgtatttg tctgaaaatt agggccagac tgttaccact cccttaagtt
      961 tgaccttaga tcactggaaa gatgtcgagc ggctcgctca caaccagtcg gtagatgtca
     1021 agaagagacg ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc
     1081 cgcgagacgg cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac
     1141 ctggcccgca tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt
     1201 ttgacccccc tccctgggtc aagccctttg tacaccctaa gcctccgcct cctcttcttc
     1261 catccgcgcc gtctctcccc cttgaacctc ctctttcgac cccgcctcaa tcctcccttt
     1321 atccagccct cactccttct ctaggcgccg gccgtcgcca ccatggtgag caagggcgag
     1381 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     1441 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     1501 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgtcc
     1561 tggggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
     1621 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     1681 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     1741 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     1801 tttagcgaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc
     1861 aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac
     1921 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     1981 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     2041 gccgccggga tcactctcgg catggacgag ctgtacaagt actcagatct gatggaaact
     2101 ttgtctttcc ccagatataa tgtagctgag attgtgattc atattcgcaa taagatctta
     2161 acaggagctg atggtaaaaa cctcaccaag aatgatcttt atccaaatcc aaagcctgaa
     2221 gtcttgcaca tgatctacat gagagcctta caaatagtct atggaattcg actggaacat
     2281 ttttacatga tgccagtgaa ctctgaagtc atgtatccac atttaatgga aggcttctta
     2341 ccattcagca atttagttac tcatctggac tcatttttgc ctatctgccg ggtgaatgac
     2401 tttgagactg ctgatattct atgtccaaaa gcaaaacgga caagtcggtt tttaagtggc
     2461 attatcaact ttattcactt cagagaagca tgccgtgaaa cgtatatgga atttctttgg
     2521 caatataaat cctctgcgga caaaatgcaa cagttaaacg ccgcacacca ggaggcatta
     2581 atgaaactgg agagacttga ttctgttcca gttgaagagc aagaagagtt caagcagctt
     2641 tcagatggaa ttcaggagct acaacaatca ctaaatcagg attttcatca aaaaacgata
     2701 gtgctgcaag agggaaattc ccaaaagaag tcaaatattt cagagaaaac caagcgtttg
     2761 aatgaactaa aattgttggt ggtttctttg aaagaaatac aagagagttt gaaaacaaaa
     2821 attgtggatt ctccagagaa gttaaagaat tataaagaaa aaatgaaaga tacggtccag
     2881 aagcttaaaa atgccagaca agaagtggtg gagaaatatg aaatctatgg agactcagtt
     2941 gactgcctgc cttcatgtca gttggaagtg cagttatatc aaaagaaaat acaggacctt
     3001 tcagataata gggaaaaatt agccagtatc ttaaaggaga gcctgaactt ggaggaccaa
     3061 attgagagtg atgagtcaga actgaagaaa ttgaagactg aagaaaattc gttcaaaaga
     3121 ctgatgattg tgaagaagga aaaacttgcc acagcacaat tcaaaataaa taagaagcat
     3181 gaagatgtta agcaatacaa acgcacagta attgaggatt gcaataaagt tcaagaaaaa
     3241 agaggtgctg tctatgaacg agtaaccaca attaatcaag aaatccaaaa aattaaactt
     3301 ggaattcaac aactaaaaga tgctgctgaa agggagaaac tgaagtccca ggaaatattt
     3361 ctaaacttga aaactgcttt ggagaaatac cacgacggta ttgaaaaggc agcagaggac
     3421 tcctatgcta agatagatga gaagacagct gaactgaaga ggaagatgtt caaaatgtca
     3481 accggatcca ccggatctag ataactgatc agaattctgc agtcgacggt acccgccagc
     3541 acagtggtcg accctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3601 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3661 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3721 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3781 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     3841 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttac
     3901 catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt
     3961 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga
     4021 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     4081 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     4141 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     4201 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     4261 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     4321 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     4381 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     4441 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
     4501 acgcccgccc cacgacccgc agcgcccgac cgaaaggagc gcacgacccc atgcatcgat
     4561 aaaataaaag attttattta gtctccagaa aaaagggggg aatgaaagac cccacctgta
     4621 ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaaat acataactga
     4681 gaatagagaa gttcagatca aggtcaggaa cagatggaac agctgaatat gggccaaaca
     4741 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggaacagctg
     4801 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     4861 cagatggtcc ccagatgcgg tccagccctc agcagtttct agagaaccat cagatgtttc
     4921 cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg
     4981 cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc
     5041 tcactcgggg cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa
     5101 accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga
     5161 gtgattgact acccgtcagc gggggtcttt catttccgac ttgtggtctc gctgccttgg
     5221 gagggtctcc tctgagtgat tgactacccg tcagcggggg tcttcacatg cagcatgtat
     5281 caaaattaat ttggtttttt ttcttaagta tttacattaa atggccatag ttgcattaat
     5341 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg cgctcttccg cttcctcgct
     5401 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5461 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5521 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5581 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5641 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5701 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5761 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5821 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5881 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5941 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6001 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6061 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6121 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6181 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6241 aaggatcttc acctagatcc ttttaaatta aaaatgaagt 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 ggcgtcaaca 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
//
