LOCUS       sequence-320116-        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"
     CDS             130..546
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     476..498
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     514..530
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      581..590
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             980..1003
                     /codon_start=1
                     /product="antigenic CD8+ T cell epitope from chicken 
                     ovalbumin"
                     /label=OVA peptide
                     /translation="SIINFEKL"
     CDS             1376..1429
                     /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             1436..2146
                     /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(1581..1599)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1849..1868
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2158..2178)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2163..2492
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2343..2478
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2502..2876
                     /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             3047..3516
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3646..3663)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3817..4405)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3897..3916)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4565..5356,5357..5425))
                     /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     5188..5207
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5426..5530)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5561..6030
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    6095..6294
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 ttcccgcctc cgtctgaatt tttgctttcg gtttggaacc gaagccgcgc gtcttgtctg
       61 ctgcagcgct gcagcatcgt tctgtgttgt ctctgtctga ctgtgtttct gtatttgtct
      121 gaaaattagg gccagactgt taccactccc ttaagtttga ccttaggtca ctggaaagat
      181 gtcgagcgga tcgctcacaa ccagtcggta gatgtcaaga agagacgttg ggttaccttc
      241 tgctctgcag aatggccaac ctttaacgtc ggatggccgc gagacggcac ctttaaccga
      301 gacctcatca cccaggttaa gatcaaggtc ttttcacctg gcccgcatgg acacccagac
      361 caggtcccct acatcgtgac ctgggaagcc ttggcttttg acccccctcc ctgggtcaag
      421 ccctttgtac accctaagcc tccgcctcct cttcctccat ccgccccgtc tctccccctt
      481 gaacctcctc gttcgacccc gcctcgatcc tccctttatc cagccctcac tccttctcta
      541 ggcgccggcc ggatcccagt gtggtggtac gtaggaattc gccaccatgg gctccatcgg
      601 cgcagcaagc atggaatttt gttttgatgt attcaaggag ctcatcaatt cctgggtaga
      661 aagtcagaca aatggaatta tcagaaatgt ccttcagcca agctccgtgg attctcaaac
      721 tgcaatggtt ctggttaatg ccattgtctt caaaggactg tgggagaaaa catttaagga
      781 tgaagacaca caagcaatgc ctttcagagt gactgagcaa gaaagcaaac ctgtgcagat
      841 gatgtaccag attggtttat ttagagtggc atcaatggct tctgagaaaa tgaagatcct
      901 ggagcttcca tttgccagtg ggacaatgag catgttggtg ctgttgcctg atgaagtctc
      961 aggccttgag cagcttgaga gtataatcaa ctttgaaaaa ctgactgaat ggaccagttc
     1021 taatgttatg gaagagagga agatcaaagt gtacttacct cgcatgaaga tggaggaaaa
     1081 atacaacctc acatctgtct taatggctat gggcattact gacgtgttta gctcttcagc
     1141 caatctgtct ggcatctcct cagcagagag cctgaagata tctcaagctg tccatgcagc
     1201 acatgcagaa atcaatgaag caggcagaga ggtggtaggg tcagcagagg ctggagtgga
     1261 tgctgcaagc gtctctgaag aatttagggc tgaccatcca ttcctcttct gtatcaagca
     1321 catcgcaacc aacgccgttc tcttctttgg cagatgtgtt tcccctggct ccggcgaggg
     1381 caggggaagt cttctaacat gcggggacgt ggaggaaaat cccggcccat tcaccatggt
     1441 gagcaagggc gaggagaata acatggccat catcaaggag ttcatgcgct tcaaggtgcg
     1501 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     1561 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc
     1621 ctgggacatc ctaaccccca acttcaccta cggctccaag gcctacgtga agcaccccgc
     1681 cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     1741 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga
     1801 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca
     1861 gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct
     1921 gaagggcgag atcaagatga ggctgaagct gaaggacggc ggccactacg acgctgaggt
     1981 caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca tcgtcggcat
     2041 caagttggac atcacctccc acaacgagga ctacaccatc gtggaactgt acgaacgcgc
     2101 cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtgagtcg accctgtgga
     2161 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2221 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     2281 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     2341 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2401 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     2461 gaggcttttt tggaggccta ggcttttgca aaaagcttac catggccaag ttgaccagtg
     2521 ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg accgaccggc
     2581 tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg gacgacgtga
     2641 ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg gcctgggtgt
     2701 gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc
     2761 gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg cgggagttcg
     2821 ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag gactgaccga
     2881 tcgataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc
     2941 tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa aaatacataa
     3001 ctgagaatag agaagttcag atcaaggtca ggaacagatg gaacagctga atatgggcca
     3061 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggaaca
     3121 gctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     3181 agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagagaa ccatcagatg
     3241 tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag
     3301 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
     3361 cccctcactc ggggcgccag tcctccgatt gactgagtcg cccgggtacc cgtgtatcca
     3421 ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct
     3481 ctgagtgatt gactacccgt cagcgggggt ctttcacatg cagcatgtat caaaattaat
     3541 ttggtttttt ttcttaagta tttacattaa atggccatag ttgcattaat gaatcggcca
     3601 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3661 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3721 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3781 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3841 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3901 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3961 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4021 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4081 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4141 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4201 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4261 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4321 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4381 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4441 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4501 cacctagatc cttttgcggc cgcaaatcaa tctaaagtat atatgagtaa acttggtctg
     4561 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4621 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4681 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4741 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4801 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4861 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4921 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4981 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5041 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5101 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5161 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5221 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5281 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5341 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5401 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5461 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5521 gggttccgcg cacatttccc cgaaaagtgc cacctgcagc ctgaatatgg gccaaacagg
     5581 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg aacagctgaa
     5641 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
     5701 gatggtcccc agatgcggtc cagccctcag cagtttctag agaaccatca gatgtttcca
     5761 gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct
     5821 tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc
     5881 actcggggcg ccagtcctcc gattgactga gtcgcccggg tacccgtgta tccaataaac
     5941 cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt
     6001 gattgactac ccgtcagcgg gggtctttca tttgggggct cgtccgggat cgggagaccc
     6061 ctgcccaggg accaccgacc caccaccggg aggcaagctg gccagcaact tatctgtgtc
     6121 tgtccgattg tctagtgtct atgactgatt ttatgcgcct gcgtcggtac tagttagcta
     6181 actagctctg tatctggcgg acccgtggtg gaactgacga gttctgaaca cccggccgca
     6241 accctgggag acgtcccagg gactttgggg gccgtttttg tggcccgacc tgaggaaggg
     6301 agtcgatgtg gaatccgacc ccgtcaggat atgtggttct ggtaggagac gagaacctaa
     6361 aacag
//
