LOCUS       sequence-272444-        6214 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 6214)
  TITLE     Cheeseman Lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6214)
  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            "Unpublished"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: pDK362
FEATURES             Location/Qualifiers
     source          1..6214
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             84..500
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     430..452
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     468..484
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             993..1010
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1020..1043
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1044..1067
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(1077..1079,1080..1082,1083..1796)
                     /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(1122..1143)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1383..1402)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1730..1751
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1941..1961)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1946..2275
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2126..2261
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2283..2681
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             2850..3319
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3449..3466)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3620..4208)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3700..3719)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4368..5159,5160..5228))
                     /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     4991..5010
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5229..5333)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5364..5833
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    5898..6097
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 gcgcgtcttg tctgctgcag cgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt
       61 ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag
      121 gtcactggaa agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac
      181 gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg
      241 gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc
      301 atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc
      361 ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc
      421 cgtctctccc ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc
      481 tcactccttc tctaggcgcc ggccggatcc tcgaggccac catgttatcc aagcaaaaat
      541 cagaggaaga gctcaaggac aagaaccagc tgttagaagc cgtcaacaag cagttgcacc
      601 agaagttgac tgaaactcag ggagagctga aggacctgac ccagaaggta gagctgctgg
      661 agaagtttcg ggacaactgt ttggcaattt tggagagcaa gggccttgat ccagctttag
      721 gcagtgagac cctggcatca cgacaagaat ccactactga tcacatggac tctatgttgc
      781 tgttagaaac tttgcaagag gagctgaagc tttttaacga aacagccaaa aagcagatgg
      841 aggagttaca ggccttaaag gtaaagctgg agatgaaaga ggagcgcgtg aggtttttgg
      901 aacagcaaac cttatgtaac aatcaagtaa atgatttaac aacagccctt aaggaaatgg
      961 agcagctatt agaaatgggc cgcggcagca gccatcatca tcatcatcac agcagcggcc
     1021 tggaagttct gttccagggg cccctggaag ttctgttcca ggggccctcc ggagggatgg
     1081 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1141 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     1201 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1261 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     1321 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1381 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1441 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1501 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     1561 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     1621 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     1681 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     1741 tggagttcgt gaccgccacc gggatcactc ttggcatgga cgagctgtac aagtaaagat
     1801 ccaccggatc tagataactg atcataatca gccataccac atttgtagag gttttacttg
     1861 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattcgcc
     1921 agcacagtgg tcgaccctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     1981 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     2041 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2101 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     2161 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     2221 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     2281 tcatggccaa gcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa
     2341 tcaacagcat ccccatctct gaagactaca gcgtcgccag cgcagctctc tctagcgacg
     2401 gccgcatctt cactggtgtc aatgtatatc attttactgg gggaccttgt gcagaactcg
     2461 tggtgctggg cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg
     2521 gaaatgagaa caggggcatc ttgagcccct gcggacggtg ccgacaggtg cttctcgatc
     2581 tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga
     2641 ttcgtgaatt gctgccctct ggttatgtgt gggagggcta agatcgataa aataaaagat
     2701 tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct
     2761 agcttaagta acgccatttt gcaaggcatg gaaaaataca taactgagaa tagagaagtt
     2821 cagatcaagg tcaggaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt
     2881 aagcagttcc tgccccggct cagggccaag aacagatgga acagctgaat atgggccaaa
     2941 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca
     3001 gatgcggtcc agccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa
     3061 ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg
     3121 ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca ctcggggcgc
     3181 cagtcctccg attgactgag tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt
     3241 tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc
     3301 cgtcagcggg ggtctttcac atgcagcatg tatcaaaatt aatttggttt tttttcttaa
     3361 gtatttacat taaatggcca tagttgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3421 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3481 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3541 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3601 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3661 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3721 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3781 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3841 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3901 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3961 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4021 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4081 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4141 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4201 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4261 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttgc
     4321 ggccgcaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4381 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4441 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4501 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4561 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4621 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4681 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     4741 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     4801 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     4861 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     4921 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     4981 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5041 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5101 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5161 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5221 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5281 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5341 ccccgaaaag tgccacctgc agcctgaata tgggccaaac aggatatctg tggtaagcag
     5401 ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat
     5461 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
     5521 gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
     5581 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
     5641 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggg gcgccagtcc
     5701 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
     5761 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
     5821 cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca gggaccaccg
     5881 acccaccacc gggaggcaag ctggccagca acttatctgt gtctgtccga ttgtctagtg
     5941 tctatgactg attttatgcg cctgcgtcgg tactagttag ctaactagct ctgtatctgg
     6001 cggacccgtg gtggaactga cgagttctga acacccggcc gcaaccctgg gagacgtccc
     6061 agggactttg ggggccgttt ttgtggcccg acctgaggaa gggagtcgat gtggaatccg
     6121 accccgtcag gatatgtggt tctggtagga gacgagaacc taaaacagtt cccgcctccg
     6181 tctgaatttt tgctttcggt ttggaaccga agcc
//
