LOCUS       sequence-152246-        7693 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 7693)
  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 7693)
  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-hygro-mTurquoise2-Nuf2
FEATURES             Location/Qualifiers
     source          1..7693
                     /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             3903..4928
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3903..4928
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     LTR             4974..5567
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5768..5785)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5939..6527)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6019..6038)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6705..7496,7497..7565))
                     /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     7328..7347
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7566..7670)
                     /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 aaaaagctta
     3901 ccatgaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg
     3961 acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg
     4021 atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag
     4081 atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca
     4141 ttggggaatt tagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt
     4201 tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggccatgg
     4261 atgcgatcgc tgcggccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag
     4321 gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt
     4381 atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg
     4441 agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg
     4501 gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg
     4561 cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg
     4621 cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc
     4681 cgcggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg
     4741 acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg
     4801 gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg
     4861 gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa
     4921 aggaatagat cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa
     4981 gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa
     5041 aatacataac tgagaataga gaagttcaga tcaaggtcag gaacagatgg aacagctgaa
     5101 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
     5161 gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc
     5221 tcagggccaa gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac
     5281 catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta
     5341 accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag
     5401 agcccacaac ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc
     5461 gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga
     5521 gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatttcc gacttgtggt
     5581 ctcgctgcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtcttcac
     5641 atgcagcatg tatcaaaatt aatttggttt tttttcttaa gtatttacat taaatggcca
     5701 tagttgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tggcgctctt
     5761 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5821 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5881 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5941 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6001 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6061 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6121 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6181 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6241 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6301 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6361 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6421 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6481 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6541 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6601 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agtttgcggc
     6661 cgcaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6721 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6781 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6841 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     6901 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     6961 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     7021 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     7081 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     7141 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     7201 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     7261 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     7321 acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     7381 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     7441 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7501 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7561 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7621 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7681 cgaaaagtgc cac
//
