LOCUS       sequence-272871-        6643 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral plasmid with SKAP insert.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6643)
  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 6643)
  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: pDK366
FEATURES             Location/Qualifiers
     source          1..6643
                     /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             942..1358
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1288..1310
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1326..1342
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             2109..2126
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2136..2159
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2160..2183
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(2193..2195,2196..2198,2199..2912)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTATGITLGMDELYK"
     primer_bind     complement(2238..2259)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2499..2518)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2846..2867
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3057..3077)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3062..3391
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3242..3377
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3474..3872
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     complement(4717..4734)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4888..5476)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4968..4987)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5655..6446,6447..6515))
                     /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     6278..6297
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6516..6620)
                     /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 caagctggcc 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 tgctgcagcg ctgcagcatc gttctgtgtt
      901 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
      961 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
     1021 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     1081 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     1141 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     1201 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     1261 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
     1321 cctcccttta tccagccctc actccttctc taggcgccgg ccggatcctc gaggccacca
     1381 tgaccagtgt ggttaagaca gtgtatagcc tgcagccccc ctctgcgctg agcggcggcc
     1441 agccggcaga cacacaaact cgggccactt ctaagagtct cttacctgtt aggtccaaag
     1501 aagtcgatgt ttccaaacag cttcattcag gaggtccaga gaatgatgtt acaaaaatca
     1561 ccaaactgag acgagagaat gggcaaatga aagctactga cactgccacc gacgataacg
     1621 tggacgatgg atatgaccct ttatccaagc aaaaatcaga ggaagagctc aaggacaaga
     1681 accagctgtt agaagccgtc aacaagcagt tgcaccagaa gttgactgaa actcagggag
     1741 agctgaagga cctgacccag aaggtagagc tgctggagaa gtttcgggac aactgtttgg
     1801 caattttgga gagcaagggc cttgatccag ctttaggcag tgagaccctg gcatcacgac
     1861 aagaatccac tactgatcac atggactcta tgttgctgtt agaaactttg caagaggagc
     1921 tgaagctttt taacgaaaca gccaaaaagc agatggagga gttacaggcc ttaaaggtaa
     1981 agctggagat gaaagaggag cgcgtgaggt ttttggaaca gcaaacctta tgtaacaatc
     2041 aagtaaatga tttaacaaca gcccttaagg aaatggagca gctattagaa atgggccgcg
     2101 gcagcagcca tcatcatcat catcacagca gcggcctgga agttctgttc caggggcccc
     2161 tggaagttct gttccagggg ccctccggag ggatggtgag caagggcgag gagctgttca
     2221 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     2281 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     2341 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     2401 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     2461 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     2521 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     2581 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     2641 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     2701 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     2761 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     2821 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccaccggga
     2881 tcactcttgg catggacgag ctgtacaagt aaagatccac cggatctaga taactgatca
     2941 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     3001 ccctgaacct gaaacataaa atgaatgcaa ttcgccagca cagtggtcga ccctgtggaa
     3061 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3121 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3181 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3241 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3301 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     3361 aggctttttt ggaggcctag gcttttgcaa aaagcttcat ggccaagcct ytgwnctmww
     3421 smagaaktar tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttcatggcca
     3481 agcctttgtc tcaagaagaa tccaccctca ttgaaagagc aacggctaca atcaacagca
     3541 tccccatctc tgaagactac agcgtcgcca gcgcagctct ctctagcgac ggccgcatct
     3601 tcactggtgt caatgtatat cattttactg ggggaccttg tgcagaactc gtggtgctgg
     3661 gcactgctgc tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga
     3721 acaggggcat cttgagcccc tgcggacggt gccgacaggt gcttctcgat ctgcatcctg
     3781 ggatcaaagc catagtgaag gacagtgatg gacagccgac ggcagttggg attcgtgaat
     3841 tgctgccctc tggttatgtg tgggagggct aagatcgata aaataaaaga ttttatttag
     3901 tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc tagcttaagt
     3961 aacgccattt tgcaaggcat ggaaaaatac ataactgaga atagagaagt tcagatcaag
     4021 gtcaggaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc
     4081 ctgccccggc tcagggccaa gaacagatgg aacagctgaa tatgggccaa acaggatatc
     4141 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc
     4201 cagccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa
     4261 atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc
     4321 ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggggcg ccagtcctcc
     4381 gattgactga gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga
     4441 cttgtggtct cgctgttcct tgggagggtc tcctctgagt gaktgwmtmc mmkwmascsk
     4501 skkkcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
     4561 tgactacccg tcagcggggg tctttcacat gcagcatgta tcaaaattaa tttggttttt
     4621 tttcttaagt atttacatta aatggccata gttgcattaa tgaatcggcc aacgcgcggg
     4681 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4741 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4801 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4861 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4921 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4981 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5041 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5101 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5161 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5221 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5281 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5341 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5401 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5461 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5521 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5581 ccttttgcgg ccgcaaatca atctaaagta tatatgwswa wmywwrgtmt rwmwgagtaa
     5641 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5701 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5761 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5821 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5881 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5941 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6001 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6061 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6121 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6181 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6241 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6301 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6361 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6421 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6481 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6541 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6601 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cac
//
