LOCUS       sequence-272443-        6472 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 6472)
  TITLE     Cheeseman Lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6472)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pDK365
FEATURES             Location/Qualifiers
     source          1..6472
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    332..531
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             732..1148
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1078..1100
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1116..1132
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1899..1916
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1926..1949
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1950..1973
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(1983..1985,1986..1988,1989..2702)
                     /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(2028..2049)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2289..2308)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2636..2657
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2847..2867)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2852..3181
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3032..3167
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3189..3587
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             3756..4225
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4355..4372)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4526..5114)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4606..4625)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5274..6065,6066..6134))
                     /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     5897..5916
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6135..6239)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             join(6270..6472,1..267)
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct
       61 cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact
      121 cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct
      181 cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
      241 tgactacccg tcagcggggg tctttcattt gggggctcgt ccgggatcgg gagacccctg
      301 cccagggacc accgacccac caccgggagg caagctggcc agcaacttat ctgtgtctgt
      361 ccgattgtct agtgtctatg actgatttta tgcgcctgcg tcggtactag ttagctaact
      421 agctctgtat ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc
      481 ctgggagacg tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt
      541 cgatgtggaa tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac
      601 agttcccgcc tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc
      661 tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt
      721 ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag
      781 atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct
      841 tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc
      901 gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag
      961 accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca
     1021 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc
     1081 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc
     1141 taggcgccgg ccggatcctc gaggccacca tgaccagtgt ggttaagaca gtgtatagcc
     1201 tgcagccccc ctctgcgctg agcggcggcc agccggcaga cacacaaact cgggccactt
     1261 ctaagagtct cttacctgtt aggtccaaag aagtcgatgt ttccaaacag cttcattcag
     1321 gaggtccaga gaatgatgtt acaaaaatca ccaaactgag acgagagaat gggcaaatga
     1381 aagctactga cactgccacc agaaggaatg tcagaaaggg atataagcct ttatccaagc
     1441 aaaaatcaga ggaagagctc aaggacaaga accagctgtt agaagccgtc aacaagcagt
     1501 tgcaccagaa gttgactgaa actcagggag agctgaagga cctgacccag aaggtagagc
     1561 tgctggagaa gtttcgggac aactgtttgg caattttgga gagcaagggc cttgatccag
     1621 ctttaggcag tgagaccctg gcatcacgac aagaatccac tactgatcac atggactcta
     1681 tgttgctgtt agaaactttg caagaggagc tgaagctttt taacgaaaca gccaaaaagc
     1741 agatggagga gttacaggcc ttaaaggtaa agctggagat gaaagaggag cgcgtgaggt
     1801 ttttggaaca gcaaacctta tgtaacaatc aagtaaatga tttaacaaca gcccttaagg
     1861 aaatggagca gctattagaa atgggccgcg gcagcagcca tcatcatcat catcacagca
     1921 gcggcctgga agttctgttc caggggcccc tggaagttct gttccagggg ccctccggag
     1981 ggatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     2041 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     2101 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     2161 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     2221 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     2281 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     2341 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     2401 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     2461 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     2521 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     2581 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     2641 tcctgctgga gttcgtgacc gccaccggga tcactcttgg catggacgag ctgtacaagt
     2701 aaagatccac cggatctaga taactgatca taatcagcca taccacattt gtagaggttt
     2761 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     2821 ttcgccagca cagtggtcga ccctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     2881 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     2941 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3001 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3061 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3121 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3181 aaagcttcat ggccaagcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg
     3241 ctacaatcaa cagcatcccc atctctgaag actacagcgt cgccagcgca gctctctcta
     3301 gcgacggccg catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag
     3361 aactcgtggt gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg
     3421 cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc
     3481 tcgatctgca tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag
     3541 ttgggattcg tgaattgctg ccctctggtt atgtgtggga gggctaagat 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 tttcacatgc agcatgtatc aaaattaatt tggttttttt
     4261 tcttaagtat ttacattaaa tggccatagt tgcattaatg aatcggccaa cgcgcgggga
     4321 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4381 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4441 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4501 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4561 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4621 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4681 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4741 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4801 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4861 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4921 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4981 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5041 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5101 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5161 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5221 ttttgcggcc gcaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5281 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5341 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5401 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5461 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5521 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5581 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5641 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     5701 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     5761 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5821 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     5881 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5941 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6001 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6061 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6121 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6181 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6241 acatttcccc gaaaagtgcc acctgcagcc tgaatatggg ccaaacagga tatctgtggt
     6301 aagcagttcc tgccccggct cagggccaag aacagatgga acagctgaat atgggccaaa
     6361 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca
     6421 gatgcggtcc agccctcagc agtttctaga gaaccatcag atgtttccag gg
//
