LOCUS       sequence-344142-        6427 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses palmytoylated GFP with a C terminal fusion of the LLO190 
            and minigenes of 2 neoantigens from the T3 tumor in genes Alg8 and 
            Lama4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6427)
  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 6427)
  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-mtGFP-llo-lama4-alg8
FEATURES             Location/Qualifiers
     source          1..6427
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    33..54
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(171..188)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(342..930)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(422..441)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1101..1892,1893..1961))
                     /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     1724..1743
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1962..2066)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2134..2152
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2190..2212)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2312..2331
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2525..2547
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2539..2556
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2540..2556
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(3730..3732,3733..3735,3736..4248)
                     /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(3775..3796)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4036..4055)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4383..4404
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4843..5246
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             5752..6342
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6367..6383)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6367..6383)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6380..6402)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6391..6407
                     /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(6415..6421,6422..6427,1..12,13..18))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
       61 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
      121 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgctcc ctcgctcact
      181 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
      241 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
      301 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
      361 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      421 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      481 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      541 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      601 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      661 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
      721 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      781 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
      841 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
      901 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
      961 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     1021 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     1081 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     1141 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     1201 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     1261 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     1321 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     1381 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     1441 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     1501 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     1561 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     1621 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     1681 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     1741 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     1801 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     1861 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     1921 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     1981 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     2041 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     2101 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     2161 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     2221 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     2281 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     2341 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     2401 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     2461 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     2521 ttttcccagt cacgacgttg taaaacgacg gccagtgaat tagtactcta gcttaagtaa
     2581 cgccattttg caaggcatgg aaaatacata actgagaata gagaagttca gatcaaggtt
     2641 aggaacagag agacagcaga atatgggcca aacaggatat ctgtggtaag cagttcctgc
     2701 cccggctcag ggccaagaac agttggaaca gcagaatatg ggccaaacag gatatctgtg
     2761 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtcccgc
     2821 cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga
     2881 ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct
     2941 gctccccgag ctcaataaaa gagcccacaa cccctcactc ggcgcgccag tcctccgata
     3001 gactgcgtcg cccgggtacc cgtattccca ataaagcctc ttgctgtttg catccgaatc
     3061 gtggactcgc tgatccttgg gagggtctcc tcagattgat tgactgccca cctcgggggt
     3121 ctttcatttg gaggttccac cgagatttgg agacccctgc ccagggacca ccgacccccc
     3181 cgccgggagg taagctggcc agcggtcgtt tcgtgtctgt ctctgtcttt gtgcgtgttt
     3241 gtgccggcat ctaatgtttg cgcctgcgtc tgtactagtt ggctaactag atctgtatct
     3301 ggcggtcccg cggaagaact gacgagttcg tattcccggc cgcagcccct gggagacgtc
     3361 ccagcggcct cgggggcccg ttttgtggcc cattctgtat cagttaacct acccgagtcg
     3421 gactttttgg agctccgcca ctgtccgagg ggtacgtggc tttgttgggg gacgagagac
     3481 agagacactt cccgcccccg tctgaatttt tgctttcggt tttacgccga aaccgcgccg
     3541 cgcgtcttgt ctgctgcagc atcgttctgt gttgtctctg tctgactgtg tttctgtatt
     3601 tgtctgaaaa ttagctcgac aaagttaagt aatagtccct ctctccaagc tcacttacag
     3661 gcggccgcca tgctctgttg catgaggcgg actaagcagg tggaaaaaaa cgatgaagac
     3721 caaaaaatta tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     3781 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     3841 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     3901 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     3961 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     4021 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     4081 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     4141 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     4201 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     4261 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     4321 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     4381 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcagctc aggcagttca
     4441 gttgaacggt ggaacgagaa gtatgctcaa gcctatccta atgtgtccgc taagattgat
     4501 tcttctggct ctagtgccgt cggcatcact tatacatgga caagactcta cgctagcgtc
     4561 ctcaccggca gtagttctgg cagcagtgat ggttttgaag taggattcaa ttttcggact
     4621 cttcaaccca acggccttct gtgataattc gagcatctta ccgccattta ttcccatatt
     4681 tgttctgttt ttcttgattt gggtatacat ttaaatgtta ataaaacaaa atggtggggc
     4741 aatcatttac attttatggg atatgtaatt actagttcag gtgtattgcc acaagacaaa
     4801 catgttaaga aactttcccg ttatttacgc tctgttcctg ttaatcaacc tctggattac
     4861 aaaatttgtg aaagattgac tgatattctt aactatgttg ctccttttac gctgtgtgga
     4921 tatgctgctt taatgcctct gtatcatgct attgcttccc gtacggcttt cgttttctcc
     4981 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtccgtcaa
     5041 cgtggcgtgg tgtgctctgt gtttgctgac gcaaccccca ctggctgggg cattgccacc
     5101 acctgtcaac tcctttctgg gactttcgct ttccccctcc cgatcgccac ggcagaactc
     5161 atcgccgcct gccttgcccg ctgctggaca ggggctaggt tgctgggcac tgataattcc
     5221 gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccaactgg
     5281 atcctgcgcg ggacgtcctt ctgctacgtc ccttcggctc tcaatccagc ggacctccct
     5341 tcccgaggcc ttctgccggt tctgcggcct ctcccgcgtc ttcgctttcg gcctccgacg
     5401 agtcggatct ccctttgggc cgcctccccg cctgtttcgc ctcggcgtcc ggtccgtgtt
     5461 gcttggtcgt cacctgtgca gaattgcgaa ccatggattc caccgtgaac tttgtctcct
     5521 ggcatgcaaa tcgtcaactt ggcatgccaa gaataattcg gatccaagct taggcctgct
     5581 cgctttcttg ctgtcccatt tctattaaag gttcctttgt tccctaagtc caactactaa
     5641 actgggggat attatgaagg gccttgagca tctggattct gcctagcgct aagcttaaca
     5701 cgagccatag atagaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga
     5761 ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaat
     5821 acataactga gaatagagaa gttcagatca aggttaggaa cagagagaca gcagaatatg
     5881 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagttg
     5941 gaacagcaga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
     6001 ggccaagaac agatggtccc cagatgcggt cccgccctca gcagtttcta gagaaccatc
     6061 agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca
     6121 atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc
     6181 cacaacccct cactcggcgc gccagtcctc cgatagactg cgtcgcccgg gtacccgtgt
     6241 tctcaataaa ccctcttgca gttgcatccg actcgtggtc tcgctgttcc ttgggagggt
     6301 ctcctctgag tgattgactg cccacctcgg gggtctttca ttctcgagca gcttggcgta
     6361 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     6421 acgagcc
//
