LOCUS       sequence-357261-        5638 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vaccine vector encoding 8 neonatigens specific for the MC38 colon 
            cancer mouse model.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5638)
  AUTHORS   Salvatori E, Lione L, Compagnone M, Pinto E, Conforti A, Ciliberto 
            G, Aurisicchio L, Palombo F
  TITLE     Neoantigen cancer vaccine augments anti-CTLA-4 efficacy.
  JOURNAL   NPJ Vaccines. 2022 Feb 2;7(1):15. doi: 10.1038/s41541-022-00433-9.
  PUBMED    35110563
REFERENCE   2  (bases 1 to 5638)
  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: "NPJ 
            Vaccines."; date: "2022-02-2"; volume: "7(1):15. doi"; pages: " 
            10.1038/s41541-022-00433-9"
COMMENT     Sequence Label: M8
FEATURES             Location/Qualifiers
     source          1..5638
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        321..700
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        701..904
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     855..875
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     901..925
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          1040..1865
                     /label=CMV intron A
                     /note="human cytomegalovirus intron A (Chapman et al., 
                     1991)"
     polyA_signal    2655..2879
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3217..3234)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3388..3976)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3468..3487)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4518..5333)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4518..5333)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5241..5260
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5325..5344
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     5611..5629
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcagattgg
      241 ctattggcca ttgcatacgt tgtatccata tcataatatg tacatttata ttggctcatg
      301 tccaacatta ccgccatgtt gacattgatt attgactagt tattaatagt aatcaattac
      361 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      421 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      481 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      541 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      601 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      661 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      721 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      781 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      841 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      901 agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca
      961 tagaagacac cgggaccgat ccagcctccg cggccgggaa cggtgcattg gaacgcggat
     1021 tccccgtgcc aagagtgacg taagtaccgc ctatagactc tataggcaca cccctttggc
     1081 tcttatgcat gctatactgt ttttggcttg gggcctatac acccccgctt ccttatgcta
     1141 taggtgatgg tatagcttag cctataggtg tgggttattg accattattg accactcccc
     1201 tattggtgac gatactttcc attactaatc cataacatgg ctctttgcca caactatctc
     1261 tattggctat atgccaatac tctgtccttc agagactgac acggactctg tatttttaca
     1321 ggatggggtc ccatttatta tttacaaatt cacatataca acaacgccgt cccccgtgcc
     1381 cgcagttttt attaaacata gcgtgggatc tccacgcgaa tctcgggtac gtgttccgga
     1441 catgggctct tctccggtag cggcggagct tccacatccg agccctggtc ccatgcctcc
     1501 agcggctcat ggtcgctcgg cagctccttg ctcctaacag tggaggccag acttaggcac
     1561 agcacaatgc ccaccaccac cagtgtgccg cacaaggccg tggcggtagg gtatgtgtct
     1621 gaaaatgagc gtggagattg ggctcgcacg gctgacgcag atggaagact taaggcagcg
     1681 gcagaagaag atgcaggcag ctgagttgtt gtattctgat aagagtcaga ggtaactccc
     1741 gttgcggtgc tgttaacggt ggagggcagt gtagtctgag cagtactcgt tgctgccgcg
     1801 cgcgccacca gacataatag ctgacagact aacagactgt tcctttccat gggtcttttc
     1861 tgcagtcacc gtccttagat caccatggat gcaatgaaga gagggctctg ctgtgtgctg
     1921 ctgctgtgtg gagcagtctt cgtttcgccc agcttaatta agggcatccc cgtgcacctg
     1981 gagctggcca gcatgaccaa catggagctg atgagcagca tcgtgcacca gcaggtgttc
     2041 cccaccagcc tggtgatcag cgccagcatc atcgtgttca acctgctgga gctggagggc
     2101 gactacaggg acgaccacat cttcagcctg ggcagggtgc tggagctgtt cagggccgcc
     2161 cagctggcca acgacgtggt gctgcagatc atggagctgt gcggcgccac caggctgctg
     2221 aggctgacca agggcaggtt cgccctgatg aacctgaagg ccctggacgt gttcggcacc
     2281 ggcgccagca gggagttcgg cgagaagagg agcatcagcg ccatcaggag ctaccagtac
     2341 gtgatgaaga tctgcgccgc ccacctgccc accgagagcg agctgagcag ctgcctgagc
     2401 aacttccact tcatgtgcgc cctggccagc tactgcatct tccaggacga caagaaggtg
     2461 ttctgcaccg agctgaggat gaccagggcc atccagcccc agatcaacgc cttcctgcag
     2521 ggcttccaca tgttcatccc ccccagcctg ggccacggcc actgggtgaa caccatggcc
     2581 ctgagcacct actacgccct gaggaccggc gccttcgagc ccgccgaggc cacctaatga
     2641 gcggccgcga tctgctgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     2701 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     2761 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     2821 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     2881 cgctgcggcc aggtgctgaa gaattgaccc ggttcctcct gggccagaaa gaagcaggca
     2941 catccccttc tctgtgacac accctgtcca cgcccctggt tcttagttcc agccccactc
     3001 ataggacact catagctcag gagggctccg ccttcaatcc cacccgctaa agtacttgga
     3061 gcggtctctc cctccctcat cagcccacca aaccaaacct agcctccaag agtgggaaga
     3121 aattaaagca agataggcta ttaagtgcag agggagagaa aatgcctcca acatgtgagg
     3181 aagtaatgag agaaatcata gaatttcttc cgcttcctcg ctcactgact cgctgcgctc
     3241 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3301 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3361 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3421 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3481 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3541 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3601 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3661 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3721 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3781 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     3841 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3901 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3961 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4021 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4081 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4141 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4201 atccatagtt gcctgactcg gggggggggg gcgctgaggt ctgcctcgtg aagaaggtgt
     4261 tgctgactca taccaggcct gaatcgcccc atcatccagc cagaaagtga gggagccacg
     4321 gttgatgaga gctttgttgt aggtggacca gttggtgatt ttgaactttt gctttgccac
     4381 ggaacggtct gcgttgtcgg gaagatgcgt gatctgatcc ttcaactcag caaaagttcg
     4441 atttattcaa caaagccgcc gtcccgtcaa gtcagcgtaa tgctctgcca gtgttacaac
     4501 caattaacca attctgatta gaaaaactca tcgagcatca aatgaaactg caatttattc
     4561 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
     4621 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt
     4681 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
     4741 tcaccatgag tgacgactga atccggtgag aatggcaaaa gcttatgcat ttctttccag
     4801 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
     4861 ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa
     4921 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt
     4981 tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg
     5041 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata
     5101 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct
     5161 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc
     5221 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg
     5281 ttggaattta atcgcggcct cgagcaagac gtttcccgtt gaatatggct cataacaccc
     5341 cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct
     5401 tgtgcaatgt aacatcagag attttgagac acaacgtggc tttccccccc cccccattat
     5461 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5521 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     5581 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
