LOCUS       sequence-158426-        6353 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  human CENP-A N-terminally labeled with LSSmOrange.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6353)
  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 6353)
  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: pTY07
FEATURES             Location/Qualifiers
     source          1..6353
                     /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      1362..1371
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1368..2075
                     /codon_start=1
                     /product="monomeric orange fluorescent protein with a large
                     Stokes shift (Shcherbakova et al., 2012)"
                     /label=LSSmOrange
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTVKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGMEASSERMYPEDGALK
                     GEDKLRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     1781..1800
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2592..2612)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2597..2926
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2777..2912
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2936..3535
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     primer_bind     complement(2936..2955)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3432..3452
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3634..4227
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4428..4445)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4599..5187)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4679..4698)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5365..6156,6157..6225))
                     /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     5988..6007
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6226..6330)
                     /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 gccggatcct cgccaccatg gtgagcaagg
     1381 gcgaggagaa taacatggcc atcatcaagg agttcatgcg cttcaaggtg cgcatggagg
     1441 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
     1501 gctttcagac cgttaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca
     1561 tcttgtcccc tcagttcacc tacggctcca aggcctacgt gaagcacccc gccgacatcc
     1621 ccgactacct caagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
     1681 aggacggcgg cgtggtgacc gtgactcagg actcctccct gcaggacggc gagttcatct
     1741 acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga
     1801 ccatgggcat ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg
     1861 aggacaagct caggctgaag ctgaaggacg gcggccacta cacctccgag gtcaagacca
     1921 cctacaaggc caagaagccc gtgcagttgc ccggcgccta catcgtcgac atcaagttgg
     1981 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc
     2041 gccactccac cggcggcatg gacgagctgt acaagggagg aacatccttc gcatctgcaa
     2101 cgaagttatc agtcgacacc atgggcccgc gccgccggag ccgaaagccc gaggccccga
     2161 ggaggcgcag cccgagcccg accccgaccc ccggcccctc ccggcggggc ccctccttag
     2221 gcgcttcctc ccatcaacac agtcggcgga gacaaggttg gctaaaggag atccgaaagc
     2281 ttcagaagag cacacacctc ttgataagga agctgccctt cagccgcctg gcaagagaaa
     2341 tatgtgttaa attcactcgt ggtgtggact tcaattggca agcccaggcc ctattggccc
     2401 tacaagaggc agcagaagca tttctagttc atctctttga ggacgcctat ctcctcacct
     2461 tacatgcagg ccgagttact ctcttcccaa aggatgtgca actggcccgg aggatccggg
     2521 gccttgagga gggactcggc taagatctcg agctcaagct tcgaattcgc cagcacagtg
     2581 gtcgaccctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     2641 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     2701 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     2761 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     2821 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     2881 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttaccatgac
     2941 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac
     3001 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca
     3061 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     3121 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     3181 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     3241 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     3301 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     3361 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     3421 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     3481 cgagtgcccg aaggaccgcg cgacctggtg catgacccgc aagcccggtg cctgacgccc
     3541 gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga ccccatgcat cgataaaata
     3601 aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg
     3661 caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac tgagaataga
     3721 gaagttcaga tcaaggtcag gaacagatgg aacagctgaa tatgggccaa acaggatatc
     3781 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag ctgaatatgg
     3841 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
     3901 tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt ttccagggtg
     3961 ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg
     4021 cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg
     4081 gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa taaaccctct
     4141 tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg
     4201 actacccgtc agcgggggtc tttcatttcc gacttgtggt ctcgctgcct tgggagggtc
     4261 tcctctgagt gattgactac ccgtcagcgg gggtcttcac atgcagcatg tatcaaaatt
     4321 aatttggttt tttttcttaa gtatttacat taaatggcca tagttgcatt aatgaatcgg
     4381 ccaacgcgcg gggagaggcg gtttgcgtat tggcgctctt ccgcttcctc gctcactgac
     4441 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4501 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4561 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4621 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4681 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4741 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4801 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4861 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4921 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4981 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     5041 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5101 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5161 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5221 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5281 ttcacctaga tccttttaaa ttaaaaatga agtttgcggc cgcaaatcaa tctaaagtat
     5341 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5401 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5461 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     5521 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5581 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5641 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5701 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5761 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5821 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5881 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5941 atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc
     6001 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6061 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6121 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6181 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6241 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6301 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cac
//
