LOCUS       sequence-344450-        6502 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses palmitoylated GFP with a C terminal fusion of the LLO190 
            and minigenes of 3 neoantigens from the MC38 tumor in genes Aatf, 
            irgq and cpne1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6502)
  AUTHORS   Veatch JR, Singhi N, Srivastava S, Szeto JL, Jesernig B, Stull SM, 
            Fitzgibbon M, Sarvothama M, Yechan-Gunja S, James SE, Riddell SR
  TITLE     A therapeutic cancer vaccine delivers antigens and adjuvants to 
            lymphoid tissues using genetically modified T cells.
  JOURNAL   J Clin Invest. 2021 Aug 16;131(16):e144195. doi: 10.1172/JCI144195.
  PUBMED    34396986
REFERENCE   2  (bases 1 to 6502)
  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: "J Clin 
            Invest."; date: "2021-08-16"; volume: "131(16):e144195. doi"; pages:
            " 10.1172/JCI144195"
COMMENT     Sequence Label: pMP71-mGFP-llo190-mc38minor
FEATURES             Location/Qualifiers
     source          1..6502
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..172
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    686..1089
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             1595..2185
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2210..2226)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2210..2226)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2223..2245)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2234..2250
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2258..2264,2265..2282,2283..2288))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2303..2324
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2441..2458)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2612..3200)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2692..2711)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3371..4162,4163..4231))
                     /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     3994..4013
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4232..4336)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4404..4422
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4460..4482)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4582..4601
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4795..4817
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4809..4826
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4810..4826
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(6000..6002,6003..6005,6006..6502,1..16)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIE"
     primer_bind     complement(6045..6066)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6306..6325)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
       61 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
      121 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
      181 cgggatcact ctcagctcag gcagttcagt tgaacggtgg aacgagaagt atgctcaagc
      241 ctatcctaat gtgtccgcta agattgattc ttctggctct agtggagatc ccagtagtcc
      301 ttattccctt cactacctgt cccctactgg tgtgaatgaa tcttccgggt cctccgcaag
      361 agatgaaact gccgcactct tgaactccgc agtgttgggg gctgcccccc tttcctcagg
      421 ctcctctaaa cttttgagtt ttatggcccc catagaccat acaacaatgt ccgatgacgc
      481 actctgagaa ttcgagcatc ttaccgccat ttattcccat atttgttctg tttttcttga
      541 tttgggtata catttaaatg ttaataaaac aaaatggtgg ggcaatcatt tacattttat
      601 gggatatgta attactagtt caggtgtatt gccacaagac aaacatgtta agaaactttc
      661 ccgttattta cgctctgttc ctgttaatca acctctggat tacaaaattt gtgaaagatt
      721 gactgatatt cttaactatg ttgctccttt tacgctgtgt ggatatgctg ctttaatgcc
      781 tctgtatcat gctattgctt cccgtacggc tttcgttttc tcctccttgt ataaatcctg
      841 gttgctgtct ctttatgagg agttgtggcc cgttgtccgt caacgtggcg tggtgtgctc
      901 tgtgtttgct gacgcaaccc ccactggctg gggcattgcc accacctgtc aactcctttc
      961 tgggactttc gctttccccc tcccgatcgc cacggcagaa ctcatcgccg cctgccttgc
     1021 ccgctgctgg acaggggcta ggttgctggg cactgataat tccgtggtgt tgtcggggaa
     1081 gctgacgtcc tttccatggc tgctcgcctg tgttgccaac tggatcctgc gcgggacgtc
     1141 cttctgctac gtcccttcgg ctctcaatcc agcggacctc ccttcccgag gccttctgcc
     1201 ggttctgcgg cctctcccgc gtcttcgctt tcggcctccg acgagtcgga tctccctttg
     1261 ggccgcctcc ccgcctgttt cgcctcggcg tccggtccgt gttgcttggt cgtcacctgt
     1321 gcagaattgc gaaccatgga ttccaccgtg aactttgtct cctggcatgc aaatcgtcaa
     1381 cttggcatgc caagaataat tcggatccaa gcttaggcct gctcgctttc ttgctgtccc
     1441 atttctatta aaggttcctt tgttccctaa gtccaactac taaactgggg gatattatga
     1501 agggccttga gcatctggat tctgcctagc gctaagctta acacgagcca tagatagaat
     1561 aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg
     1621 gcaagctagc ttaagtaacg ccattttgca aggcatggaa aatacataac tgagaataga
     1681 gaagttcaga tcaaggttag gaacagagag acagcagaat atgggccaaa caggatatct
     1741 gtggtaagca gttcctgccc cggctcaggg ccaagaacag ttggaacagc agaatatggg
     1801 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt
     1861 ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc
     1921 cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc
     1981 ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg
     2041 cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tgttctcaat aaaccctctt
     2101 gcagttgcat ccgactcgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga
     2161 ctgcccacct cgggggtctt tcattctcga gcagcttggc gtaatcatgg tcatagctgt
     2221 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     2281 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     2341 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     2401 cggggagagg cggtttgcgt attgggcgct cttccgctcc ctcgctcact gactcgctgc
     2461 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2521 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2581 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2641 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2701 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2761 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2821 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2881 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2941 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3001 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3061 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3121 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3181 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3241 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3301 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3361 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3421 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3481 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3541 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3601 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3661 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3721 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3781 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3841 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     3901 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     3961 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4021 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4081 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4141 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4201 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4261 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4321 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     4381 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     4441 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     4501 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     4561 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     4621 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat
     4681 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     4741 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     4801 cacgacgttg taaaacgacg gccagtgaat tagtactcta gcttaagtaa cgccattttg
     4861 caaggcatgg aaaatacata actgagaata gagaagttca gatcaaggtt aggaacagag
     4921 agacagcaga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
     4981 ggccaagaac agttggaaca gcagaatatg ggccaaacag gatatctgtg gtaagcagtt
     5041 cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtcccgc cctcagcagt
     5101 ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct
     5161 tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag
     5221 ctcaataaaa gagcccacaa cccctcactc ggcgcgccag tcctccgata gactgcgtcg
     5281 cccgggtacc cgtattccca ataaagcctc ttgctgtttg catccgaatc gtggactcgc
     5341 tgatccttgg gagggtctcc tcagattgat tgactgccca cctcgggggt ctttcatttg
     5401 gaggttccac cgagatttgg agacccctgc ccagggacca ccgacccccc cgccgggagg
     5461 taagctggcc agcggtcgtt tcgtgtctgt ctctgtcttt gtgcgtgttt gtgccggcat
     5521 ctaatgtttg cgcctgcgtc tgtactagtt ggctaactag atctgtatct ggcggtcccg
     5581 cggaagaact gacgagttcg tattcccggc cgcagcccct gggagacgtc ccagcggcct
     5641 cgggggcccg ttttgtggcc cattctgtat cagttaacct acccgagtcg gactttttgg
     5701 agctccgcca ctgtccgagg ggtacgtggc tttgttgggg gacgagagac agagacactt
     5761 cccgcccccg tctgaatttt tgctttcggt tttacgccga aaccgcgccg cgcgtcttgt
     5821 ctgctgcagc atcgttctgt gttgtctctg tctgactgtg tttctgtatt tgtctgaaaa
     5881 ttagctcgac aaagttaagt aatagtccct ctctccaagc tcacttacag gcggccgcca
     5941 tgctctgttg catgaggcgg actaagcagg tggaaaaaaa cgatgaagac caaaaaatta
     6001 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     6061 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     6121 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     6181 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     6241 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     6301 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     6361 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     6421 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     6481 gcatcaaggt gaacttcaag at
//
