LOCUS       sequence-65995-e        6542 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  visualizes chromosomes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6542)
  AUTHORS   Kiyomitsu T, Cheeseman IM
  TITLE     Cortical dynein and asymmetric membrane elongation coordinately 
            position the spindle in anaphase.
  JOURNAL   Cell. 2013 Jul 18;154(2):391-402. doi: 10.1016/j.cell.2013.06.010.
  PUBMED    23870127
REFERENCE   2  (bases 1 to 6542)
  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"; doi: 
            "10.1016/j.cell.2013.06"; journalName: "Cell"; date: "2013-07-18- 
            18"; volume: "154"; issue: "2"; pages: "391-402"
COMMENT     Sequence Label: pTK168_mCherry-H2B
FEATURES             Location/Qualifiers
     source          1..6542
                     /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      1384..1393
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1390..2097
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1535..1553)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1803..1822
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2143..2163
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2176..2220
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             2245..2625
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKDGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAVTKYTSAK"
     primer_bind     complement(2781..2801)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2786..3115
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2966..3101
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3125..3724
                     /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(3125..3144)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3621..3641
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3823..4416
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4617..4634)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4788..5376)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4868..4887)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5554..6345,6346..6414))
                     /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     6177..6196
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6415..6519)
                     /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 gccggatccc agtgtggtgg tacgctaccg
     1381 gtcgccacca tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg
     1441 cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
     1501 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc
     1561 cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac
     1621 gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag
     1681 tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc
     1741 ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac
     1801 ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc
     1861 gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac
     1921 tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc
     1981 tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa
     2041 cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtcc
     2101 ggaggggatc ggtggagctc caccggtggc ggccgctcta gagagaatct ttattttcag
     2161 ggcgccgcca aattcaaaga aaccgctgct gctaaattcg aacgccagca catggacagc
     2221 ggaggtggag gtactagtct cgagatgcca gagccagcga agtctgctcc cgccccgaaa
     2281 aagggctcca agaaggcggt gactaaggcg cagaagaaag acggcaagaa gcgcaagcgc
     2341 agccgcaagg agagctattc catctatgtg tacaaggttc tgaagcaggt ccaccctgac
     2401 accggcattt cgtccaaggc catgggcatc atgaattcgt ttgtgaacga cattttcgag
     2461 cgcatcgcag gtgaggcttc ccgcctggcg cattacaaca agcgctcgac catcacctcc
     2521 agggagatcc agacggccgt gcgcctgctg ctgcctgggg agttggccaa gcacgccgtg
     2581 tccgagggta ctaaggccgt caccaagtac accagcgcta agtaaccgcg ggcccgggat
     2641 ccaccggatc tagataactg atcataatca gccataccac atttgtagag gttttacttg
     2701 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattcgcc
     2761 agcacagtgg tcgaccctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     2821 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     2881 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2941 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3001 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3061 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3121 taccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc
     3181 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc
     3241 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
     3301 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
     3361 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
     3421 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
     3481 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
     3541 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     3601 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     3661 cgccgacgtc gagtgcccga aggaccgcgc gacctggtgc atgacccgca agcccggtgc
     3721 ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc
     3781 gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg
     3841 taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact
     3901 gagaatagag aagttcagat caaggtcagg aacagatgga acagctgaat atgggccaaa
     3961 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggaacagc
     4021 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     4081 aacagatggt ccccagatgc ggtccagccc tcagcagttt ctagagaacc atcagatgtt
     4141 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
     4201 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
     4261 cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg tgtatccaat
     4321 aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct
     4381 gagtgattga ctacccgtca gcgggggtct ttcatttccg acttgtggtc tcgctgcctt
     4441 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtcttcaca tgcagcatgt
     4501 atcaaaatta atttggtttt ttttcttaag tatttacatt aaatggccat agttgcatta
     4561 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt ggcgctcttc cgcttcctcg
     4621 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4681 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4741 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4801 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4861 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4921 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4981 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5041 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5101 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5161 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5221 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5281 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5341 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5401 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5461 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gtttgcggcc gcaaatcaat
     5521 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5581 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5641 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5701 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5761 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5821 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5881 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5941 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6001 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6061 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6121 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa
     6181 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6241 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6301 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6361 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6421 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6481 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6541 ac
//
