LOCUS       sequence-272655-        6699 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral plasmid expressing LAP-Clasp C terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6699)
  AUTHORS   Kern DM, Nicholls PK, Page DC, Cheeseman IM
  TITLE     A mitotic SKAP isoform regulates spindle positioning at astral 
            microtubule plus ends.
  JOURNAL   J Cell Biol. 2016 May 9;213(3):315-28. doi: 10.1083/jcb.201510117. 
            Epub 2016 May 2.
  PUBMED    27138257
REFERENCE   2  (bases 1 to 6699)
  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: 
            "10.1083/jcb.201510117"; journalName: "J Cell Biol"; date: 
            "2016-05-9- 9"; volume: "213"; issue: "3"; pages: "315-28"
COMMENT     Sequence Label: pDK197
FEATURES             Location/Qualifiers
     source          1..6699
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    309..508
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             709..1125
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1055..1077
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1093..1109
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1160..1169
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1166..1168,1169..1171,1172..1882)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1166..1168,1169..1171,1172..1882)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1211..1232)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1472..1491)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1819..1840
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1928..1948
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1961..2005
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     RBS             2458..2466
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(3051..3071)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3056..3385
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3236..3371
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3393..3791
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             3960..4429
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4559..4576)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4730..5318)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4810..4829)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5478..6269,6270..6338))
                     /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     6101..6120
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6339..6443)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             join(6474..6699,1..244)
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc
       61 tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga
      121 ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg
      181 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct
      241 ttcatttggg ggctcgtccg ggatcgggag acccctgccc agggaccacc gacccaccac
      301 cgggaggcaa gctggccagc aacttatctg tgtctgtccg attgtctagt gtctatgact
      361 gattttatgc gcctgcgtcg gtactagtta gctaactagc tctgtatctg gcggacccgt
      421 ggtggaactg acgagttctg aacacccggc cgcaaccctg ggagacgtcc cagggacttt
      481 gggggccgtt tttgtggccc gacctgagga agggagtcga tgtggaatcc gaccccgtca
      541 ggatatgtgg ttctggtagg agacgagaac ctaaaacagt tcccgcctcc gtctgaattt
      601 ttgctttcgg tttggaaccg aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt
      661 ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg aaaattaggg ccagactgtt
      721 accactccct taagtttgac cttaggtcac tggaaagatg tcgagcggat cgctcacaac
      781 cagtcggtag atgtcaagaa gagacgttgg gttaccttct gctctgcaga atggccaacc
      841 tttaacgtcg gatggccgcg agacggcacc tttaaccgag acctcatcac ccaggttaag
      901 atcaaggtct tttcacctgg cccgcatgga cacccagacc aggtccccta catcgtgacc
      961 tgggaagcct tggcttttga cccccctccc tgggtcaagc cctttgtaca ccctaagcct
     1021 ccgcctcctc ttcctccatc cgccccgtct ctcccccttg aacctcctcg ttcgaccccg
     1081 cctcgatcct ccctttatcc agccctcact ccttctctag gcgccggccg gatcccagtg
     1141 tggtggtacg ctaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt
     1201 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     1261 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     1321 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     1381 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     1441 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     1501 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     1561 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     1621 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
     1681 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     1741 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc
     1801 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     1861 cggcatggac gagctgtaca agtccggagg ggatcggtgg agctccaccg gtggcggccg
     1921 ctctagagag aatctttatt ttcagggcgc cgccaaattc aaagaaaccg ctgctgctaa
     1981 attcgaacgc cagcacatgg acagcggagg tggaggtact agtctcgagg ggccgcgggc
     2041 gcgagactac aacccgtacc cctactcaga tgccatcaac acctacgaca agaccgccct
     2101 gaaagaggct gtgttcgatg acgacatgga gcagcttcga gacgtgccca tcgaccattc
     2161 tgacctggtg gctgaccttc tgaaagagct gtccaaccac aatgagcgag tggaggaacg
     2221 gaagggagcc ctgctggagc tgctcaagat cacgcgggaa gacagccttg gtgtctggga
     2281 ggagcacttc aagaccattc tgctcctgct gctggagacc cttggagaca aagaccattc
     2341 aattcgagca ctggcgttaa gagttttgag ggaaattctg agaaatcaac cagcaagatt
     2401 taaaaactac gccgagctga cgattatgaa gactctggaa gcccacaaag actcccataa
     2461 ggaggtggtg agagcggctg aggaggctgc gtccacactg gccagttcca tccacccgga
     2521 gcagtgcatc aaggtgctct gccccatcat ccagacggcc gactacccca tcaaccttgc
     2581 tgccatcaag atgcagacca aagtcgtcga gaggatcgca aaggagtcat tgctgcagct
     2641 ccttgtcgac atcatcccag gcttgctgca gggttatgac aacaccgaaa gtagtgtgcg
     2701 taaggccagc gtgttttgct tagtggcaat ttattccgta atcggagaag acctgaaacc
     2761 tcaccttgca cagctcacag ggagcaagat gaagctacta aacttataca taaagagggc
     2821 ccagaccacc aacagcaaca gcagctcctc ctccgatgtc tccacgcaca gctaagaatt
     2881 ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc tagataactg atcataatca
     2941 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     3001 acctgaaaca taaaatgaat gcaattcgcc agcacagtgg tcgaccctgt ggaatgtgtg
     3061 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3121 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3181 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3241 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3301 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     3361 ttttggaggc ctaggctttt gcaaaaagct tcatggccaa gcctttgtct caagaagaat
     3421 ccaccctcat tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca
     3481 gcgtcgccag cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc
     3541 attttactgg gggaccttgt gcagaactcg tggtgctggg cactgctgct gctgcggcag
     3601 ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct
     3661 gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg
     3721 acagtgatgg acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt
     3781 gggagggcta agatcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat
     3841 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     3901 gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag atggaacagc
     3961 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     4021 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     4081 cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga
     4141 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
     4201 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
     4261 aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt
     4321 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt
     4381 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcac atgcagcatg
     4441 tatcaaaatt aatttggttt tttttcttaa gtatttacat taaatggcca tagttgcatt
     4501 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4561 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4621 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4681 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4741 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4801 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4861 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4921 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4981 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5041 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5101 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5161 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5221 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5281 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5341 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5401 caaaaaggat cttcacctag atccttttgc ggccgcaaat caatctaaag tatatatgag
     5461 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5521 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5581 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5641 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5701 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5761 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5821 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5881 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5941 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6001 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6061 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6121 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6181 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6241 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6301 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6361 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6421 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgc agcctgaata
     6481 tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga
     6541 tggaacagct gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc
     6601 agggccaaga acagatggtc cccagatgcg gtccagccct cagcagtttc tagagaacca
     6661 tcagatgttt ccagggtgcc ccaaggacct gaaatgacc
//
