LOCUS       sequence-320119-        7137 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of cOVA and mStrawberry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7137)
  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 7137)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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_Hyg_cOVA_T2A_mStrawberry
FEATURES             Location/Qualifiers
     source          1..7137
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             98..567
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    632..831
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1032..1448
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1378..1400
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1416..1432
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1483..1492
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1882..1905
                     /codon_start=1
                     /product="antigenic CD8+ T cell epitope from chicken 
                     ovalbumin"
                     /label=OVA peptide
                     /translation="SIINFEKL"
     CDS             2278..2331
                     /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             2338..3048
                     /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(2483..2501)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2751..2770
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3060..3080)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3065..3394
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3245..3380
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3468..4493
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3468..4493
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     primer_bind     5010..5029
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     LTR             5291..5760
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(join(6239..7030,7031..7099))
                     /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     6862..6881
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7100..7137,1..67))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
       61 ttccgcgcac atttccccga aaagtgccac ctgcagcctg aatatgggcc aaacaggata
      121 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac agctgaatat
      181 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
      241 ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat gtttccaggg
      301 tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct
      361 cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact
      421 cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct
      481 cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
      541 tgactacccg tcagcggggg tctttcattt gggggctcgt ccgggatcgg gagacccctg
      601 cccagggacc accgacccac caccgggagg caagctggcc agcaacttat ctgtgtctgt
      661 ccgattgtct agtgtctatg actgatttta tgcgcctgcg tcggtactag ttagctaact
      721 agctctgtat ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc
      781 ctgggagacg tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt
      841 cgatgtggaa tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac
      901 agttcccgcc tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc
      961 tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt
     1021 ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag
     1081 atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct
     1141 tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc
     1201 gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag
     1261 accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca
     1321 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc
     1381 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc
     1441 taggcgccgg ccggatccca gtgtggtggt acgtaggaat tcgccaccat gggctccatc
     1501 ggcgcagcaa gcatggaatt ttgttttgat gtattcaagg agctcatcaa ttcctgggta
     1561 gaaagtcaga caaatggaat tatcagaaat gtccttcagc caagctccgt ggattctcaa
     1621 actgcaatgg ttctggttaa tgccattgtc ttcaaaggac tgtgggagaa aacatttaag
     1681 gatgaagaca cacaagcaat gcctttcaga gtgactgagc aagaaagcaa acctgtgcag
     1741 atgatgtacc agattggttt atttagagtg gcatcaatgg cttctgagaa aatgaagatc
     1801 ctggagcttc catttgccag tgggacaatg agcatgttgg tgctgttgcc tgatgaagtc
     1861 tcaggccttg agcagcttga gagtataatc aactttgaaa aactgactga atggaccagt
     1921 tctaatgtta tggaagagag gaagatcaaa gtgtacttac ctcgcatgaa gatggaggaa
     1981 aaatacaacc tcacatctgt cttaatggct atgggcatta ctgacgtgtt tagctcttca
     2041 gccaatctgt ctggcatctc ctcagcagag agcctgaaga tatctcaagc tgtccatgca
     2101 gcacatgcag aaatcaatga agcaggcaga gaggtggtag ggtcagcaga ggctggagtg
     2161 gatgctgcaa gcgtctctga agaatttagg gctgaccatc cattcctctt ctgtatcaag
     2221 cacatcgcaa ccaacgccgt tctcttcttt ggcagatgtg tttcccctgg ctccggcgag
     2281 ggcaggggaa gtcttctaac atgcggggac gtggaggaaa atcccggccc attcaccatg
     2341 gtgagcaagg gcgaggagaa taacatggcc atcatcaagg agttcatgcg cttcaaggtg
     2401 cgcatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
     2461 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc
     2521 gcctgggaca tcctaacccc caacttcacc tacggctcca aggcctacgt gaagcacccc
     2581 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
     2641 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
     2701 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
     2761 cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc
     2821 ctgaagggcg agatcaagat gaggctgaag ctgaaggacg gcggccacta cgacgctgag
     2881 gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta catcgtcggc
     2941 atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaact gtacgaacgc
     3001 gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtgagt cgaccctgtg
     3061 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3121 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3181 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3241 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3301 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     3361 aggaggcttt tttggaggcc taggcttttg caaaaagctt gccagctggg gcgccctctg
     3421 gtaaggttgg gaagccctgc agatctgatc ccggggggca atgagatatg aaaaagcctg
     3481 aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc
     3541 tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg
     3601 gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc
     3661 ggcactttgc atcggccgcg ctcccgattc cggaagtgct tgacattggg gaatttagcg
     3721 agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg
     3781 aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg
     3841 ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca
     3901 ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg
     3961 tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg
     4021 ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc
     4081 tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt
     4141 cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc
     4201 agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt
     4261 atatgctccg cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg
     4321 atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg
     4381 ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt gtagaagtac
     4441 tcgccgatag tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa tagagtagat
     4501 gccgaccggg atctatcgat gcggcgggtg tggtggttac gcgtaatctg ctgcttgcaa
     4561 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4621 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     4681 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4741 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4801 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4861 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4921 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4981 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5041 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5101 ctatggaaaa acgccagcaa cgcatcgata aaataaaaga ttttatttag tctccagaaa
     5161 aaggggggaa tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt
     5221 tgcaaggcat ggaaaaatac ataactgaga atagagaagt tcagatcaag gtcaggaaca
     5281 gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc
     5341 tcagggccaa gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc
     5401 agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag
     5461 cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt
     5521 gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc
     5581 cgagctcaat aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga
     5641 gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct
     5701 cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca
     5761 catgcagcat gtatcaaaat taatttggtt ttttttctta agtatttaca ttaaatggcc
     5821 atagtgcggc cgcaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5881 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5941 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6001 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagms
     6061 rrycksaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6121 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6181 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6241 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6301 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6361 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6421 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6481 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6541 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6601 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6661 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6721 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6781 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6841 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6901 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6961 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7021 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7081 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagg
//
