LOCUS       sequence-313501-        6365 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of cOVA and mStrawberry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6365)
  AUTHORS   Yamamoto K, Venida A, Yano J, Biancur DE, Kakiuchi M, Gupta S, Sohn 
            ASW, Mukhopadhyay S, Lin EY, Parker SJ, Banh RS, Paulo JA, Wen KW, 
            Debnath J, Kim GE, Mancias JD, Fearon DT, Perera RM, Kimmelman AC
  TITLE     Autophagy promotes immune evasion of pancreatic cancer by degrading 
            MHC-I.
  JOURNAL   Nature. 2020 May;581(7806):100-105. doi: 10.1038/s41586-020-2229-5. 
            Epub 2020 Apr 22.
  PUBMED    32376951
REFERENCE   2  (bases 1 to 6365)
  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: 
            "10.1038/s41586-020-2229-5"; journalName: "Nature"; date: "2020-05";
            volume: "581"; issue: "7806"; pages: "100-105"
COMMENT     Sequence Label: pBabe_Zeo_cOVA_T2A_mStrawberry
FEATURES             Location/Qualifiers
     source          1..6365
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             9..478
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    543..742
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             943..1359
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1289..1311
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1327..1343
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1394..1403
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1793..1816
                     /codon_start=1
                     /product="antigenic CD8+ T cell epitope from chicken 
                     ovalbumin"
                     /label=OVA peptide
                     /translation="SIINFEKL"
     CDS             2189..2242
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2249..2959
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent 
                     protein"
                     /label=mStrawberry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILTPNFTYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKMRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYIVGIKLDITSHNEDYTIVELYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2394..2412)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2662..2681
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2971..2991)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2976..3305
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3156..3291
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3315..3689
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     LTR             3860..4329
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4459..4476)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4630..5218)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4710..4729)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5378..6169,6170..6238))
                     /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     6001..6020
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6239..6343)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cctgcagcct gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc
       61 agggccaaga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
      121 ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtcca gccctcagca
      181 gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
      241 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
      301 agctcaataa aagagcccac aacccctcac tcggggcgcc agtcctccga ttgactgagt
      361 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg
      421 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatt
      481 tgggggctcg tccgggatcg ggagacccct gcccagggac caccgaccca ccaccgggag
      541 gcaagctggc cagcaactta tctgtgtctg tccgattgtc tagtgtctat gactgatttt
      601 atgcgcctgc gtcggtacta gttagctaac tagctctgta tctggcggac ccgtggtgga
      661 actgacgagt tctgaacacc cggccgcaac cctgggagac gtcccaggga ctttgggggc
      721 cgtttttgtg gcccgacctg aggaagggag tcgatgtgga atccgacccc gtcaggatat
      781 gtggttctgg taggagacga gaacctaaaa cagttcccgc ctccgtctga atttttgctt
      841 tcggtttgga accgaagccg cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt
      901 tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt agggccagac tgttaccact
      961 cccttaagtt tgaccttagg tcactggaaa gatgtcgagc ggatcgctca caaccagtcg
     1021 gtagatgtca agaagagacg ttgggttacc ttctgctctg cagaatggcc aacctttaac
     1081 gtcggatggc cgcgagacgg cacctttaac cgagacctca tcacccaggt taagatcaag
     1141 gtcttttcac ctggcccgca tggacaccca gaccaggtcc cctacatcgt gacctgggaa
     1201 gccttggctt ttgacccccc tccctgggtc aagccctttg tacaccctaa gcctccgcct
     1261 cctcttcctc catccgcccc gtctctcccc cttgaacctc ctcgttcgac cccgcctcga
     1321 tcctcccttt atccagccct cactccttct ctaggcgccg gccggatccc agtgtggtgg
     1381 tacgtaggaa ttcgccacca tgggctccat cggcgcagca agcatggaat tttgttttga
     1441 tgtattcaag gagctcatca attcctgggt agaaagtcag acaaatggaa ttatcagaaa
     1501 tgtccttcag ccaagctccg tggattctca aactgcaatg gttctggtta atgccattgt
     1561 cttcaaagga ctgtgggaga aaacatttaa ggatgaagac acacaagcaa tgcctttcag
     1621 agtgactgag caagaaagca aacctgtgca gatgatgtac cagattggtt tatttagagt
     1681 ggcatcaatg gcttctgaga aaatgaagat cctggagctt ccatttgcca gtgggacaat
     1741 gagcatgttg gtgctgttgc ctgatgaagt ctcaggcctt gagcagcttg agagtataat
     1801 caactttgaa aaactgactg aatggaccag ttctaatgtt atggaagaga ggaagatcaa
     1861 agtgtactta cctcgcatga agatggagga aaaatacaac ctcacatctg tcttaatggc
     1921 tatgggcatt actgacgtgt ttagctcttc agccaatctg tctggcatct cctcagcaga
     1981 gagcctgaag atatctcaag ctgtccatgc agcacatgca gaaatcaatg aagcaggcag
     2041 agaggtggta gggtcagcag aggctggagt ggatgctgca agcgtctctg aagaatttag
     2101 ggctgaccat ccattcctct tctgtatcaa gcacatcgca accaacgccg ttctcttctt
     2161 tggcagatgt gtttcccctg gctccggcga gggcagggga agtcttctaa catgcgggga
     2221 cgtggaggaa aatcccggcc cattcaccat ggtgagcaag ggcgaggaga ataacatggc
     2281 catcatcaag gagttcatgc gcttcaaggt gcgcatggag ggctccgtga acggccacga
     2341 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     2401 gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctaaccc ccaacttcac
     2461 ctacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc
     2521 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
     2581 cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg
     2641 caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     2701 ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaaga tgaggctgaa
     2761 gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc
     2821 cgtgcagctg cccggcgcct acatcgtcgg catcaagttg gacatcacct cccacaacga
     2881 ggactacacc atcgtggaac tgtacgaacg cgccgagggc cgccactcca ccggcggcat
     2941 ggacgagctg tacaagtgag tcgaccctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3001 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3061 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3121 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3181 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3241 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3301 gcaaaaagct taccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg
     3361 tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg
     3421 acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc
     3481 aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg
     3541 ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg
     3601 agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg
     3661 tgcacttcgt ggccgaggag caggactgac cgatcgataa aataaaagat tttatttagt
     3721 ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta
     3781 acgccatttt gcaaggcatg gaaaaataca taactgagaa tagagaagtt cagatcaagg
     3841 tcaggaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc
     3901 tgccccggct cagggccaag aacagatgga acagctgaat atgggccaaa caggatatct
     3961 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc
     4021 agccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa
     4081 tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct
     4141 tctgctcccc gagctcaata aaagagccca caacccctca ctcggggcgc cagtcctccg
     4201 attgactgag tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac
     4261 ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg
     4321 ggtctttcac atgcagcatg tatcaaaatt aatttggttt tttttcttaa gtatttacat
     4381 taaatggcca tagttgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4441 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4501 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4561 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4621 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4681 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4741 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4801 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4861 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4921 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4981 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5041 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     5101 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5161 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5221 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5281 gattttggtc atgagattat caaaaaggat cttcacctag atccttttgc ggccgcaaat
     5341 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5401 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5461 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5521 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5581 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5641 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5701 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5761 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5821 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5881 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5941 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6001 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6061 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6121 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6181 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6241 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6301 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6361 tgcca
//
