LOCUS       sequence-122621-        4666 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For bacterial expression of recombinant human PRC1 tagged with His..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4666)
  AUTHORS   Nixon FM, Gutierrez-Caballero C, Hood FE, Booth DG, Prior IA, Royle 
            SJ
  TITLE     The mesh is a network of microtubule connectors that stabilizes 
            individual kinetochore fibers of the mitotic spindle.
  JOURNAL   Elife. 2015 Jun 19;4. doi: 10.7554/eLife.07635.
  PUBMED    26090906
REFERENCE   2  (bases 1 to 4666)
  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: "2015-06-19"; volume: "4. doi"; pages: " 10.7554/eLife.07635"
COMMENT     Sequence Label: His6-PRC1
FEATURES             Location/Qualifiers
     source          1..4666
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             70..92
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             112..129
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2067..2085)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2081..2128
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2225..2680
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2312..2331)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2522..2543
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2706..2810
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2811..2879,2880..3671)
                     /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     complement(3029..3048)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3842..4430
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4331..4350
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4584..4601
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcggggttc tcatcatcat
      121 catcatcatg gtatggctag catgaggaga agtgaggtgc tggcggagga gtccatagta
      181 tgtctgcaga aagccctaaa tcaccttcgg gaaatatggg agctaattgg gattccagag
      241 gaccagcggt tacaaagaac tgaggtggta aagaagcata tcaaggaact cctggatatg
      301 atgattgctg aagaggaaag cctgaaggaa agactcatca aaagcatatc cgtctgtcag
      361 aaagagctga acactctgtg cagcgagtta catgttgagc catttcagga agaaggagag
      421 acaaccatct tgcaactaga aaaagatttg cgcacccaag tggaattgat gcgaaaacag
      481 aaaaaggaga gaaaacagga actgaagcta cttcaagagc aagatcaaga actgtgcgaa
      541 attctttgta tgccccacta tgatattgac agtgcctcag tgcccagctt agaagagctg
      601 aaccagttca ggcaacatgt gacaactttg agggaaacaa aggcttctag gcgtgaggag
      661 tttgtcagta taaagagaca gatcatactg tgtatggaag aattagacca caccccagac
      721 acaagctttg aaagagatgt ggtgtgtgaa gacgaagatg ccttttgttt gtctttggag
      781 aatattgcaa cactacaaaa gttgctacgg cagctggaaa tgcagaaatc acaaaatgaa
      841 gcagtgtgtg aggggctgcg tactcaaatc cgagagctct gggacaggtt gcaaatacct
      901 gaagaagaaa gagaagctgt ggccaccatt atgtctgggt caaaggccaa ggtccggaaa
      961 gcgctgcaat tagaagtgga tcggttggaa gaactgaaaa tgcaaaacat gaagaaagtg
     1021 attgaggcaa ttcgagtgga gctggttcag tactgggacc agtgctttta tagccaggag
     1081 cagagacaag cttttgcccc tttctgtgct gaggactaca cagaaagtct gctccagctc
     1141 cacgatgctg agattgtgcg gttaaaaaac tactatgaag ttcacaagga actctttgaa
     1201 ggtgtccaga agtgggaaga aacctggagg cttttcttag agtttgagag aaaagcttca
     1261 gatccaaatc gatttacaaa ccgaggagga aatcttctaa aagaagaaaa acaacgagcc
     1321 aagctccaga aaatgctgcc caagctggaa gaagagttga aggcacgaat tgaattgtgg
     1381 gaacaggaac attcaaaggc atttatggtg aatgggcaga aattcatgga gtatgtggca
     1441 gaacaatggg agatgcatcg attggagaaa gagagagcca agcaggaaag acaactgaag
     1501 aacaaaaaac agacagagac agagatgctg tatggcagcg ctcctcgaac acctagcaag
     1561 cggcgaggac tggctcccaa tacaccgggc aaagcacgta agctgaacac taccaccatg
     1621 tccaatgcta cggccaatag tagcattcgg cctatctttg gagggacagt ctaccactcc
     1681 cccgtgtctc gacttcctcc ttctggcagc aagccagtcg ctgcttccac ctgttcaggg
     1741 aagaaaacac cccgtactgg caggcatgga gccaacaagg agaacctgga gctcaacggc
     1801 agcatcctga gtggtgggta ccctggctcg gcccccctcc agcgcaactt cagcattaat
     1861 tctgttgcca gcacctattc tgagtttgcg aaggatccgt ccctctctga cagttccact
     1921 gttgggcttc agcgagaact ttcaaaggct tccaaatctg atgctacttc tggaatcctc
     1981 aattcaacca acatccagtc ctgagaattc gaagcttgat ccggctgcta acaaagcccg
     2041 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     2101 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gatctggcgt
     2161 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     2221 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2281 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2341 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2401 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     2461 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2521 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     2581 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2641 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
     2701 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2761 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     2821 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2881 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     2941 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     3001 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     3061 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     3121 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     3181 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     3241 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     3301 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     3361 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     3421 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     3481 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     3541 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3601 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3661 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     3721 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     3781 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3841 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3901 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3961 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     4021 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4081 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4141 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4201 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4261 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4321 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4381 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4441 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     4501 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     4561 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     4621 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcag
//
