LOCUS       sequence-344456-        6361 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses palmitoylated GFP with a C terminal fusion of the LLO190 
            epitope and the human hPMEL epitope.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6361)
  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 6361)
  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-LLO-hPMEL
FEATURES             Location/Qualifiers
     source          1..6361
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(786..788,789..791,792..1304)
                     /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(831..852)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1092..1111)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1439..1460
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1833..2236
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             2742..3332
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3357..3373)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3357..3373)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3370..3392)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3381..3397
                     /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(3405..3411,3412..3429,3430..3435))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3450..3471
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3588..3605)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3759..4347)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3839..3858)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4518..5309,5310..5378))
                     /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     5141..5160
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5379..5483)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5551..5569
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5607..5629)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5729..5748
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5942..5964
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5956..5973
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5957..5973
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgatagact
       61 gcgtcgcccg ggtacccgta ttcccaataa agcctcttgc tgtttgcatc cgaatcgtgg
      121 actcgctgat ccttgggagg gtctcctcag attgattgac tgcccacctc gggggtcttt
      181 catttggagg ttccaccgag atttggagac ccctgcccag ggaccaccga cccccccgcc
      241 gggaggtaag ctggccagcg gtcgtttcgt gtctgtctct gtctttgtgc gtgtttgtgc
      301 cggcatctaa tgtttgcgcc tgcgtctgta ctagttggct aactagatct gtatctggcg
      361 gtcccgcgga agaactgacg agttcgtatt cccggccgca gcccctggga gacgtcccag
      421 cggcctcggg ggcccgtttt gtggcccatt ctgtatcagt taacctaccc gagtcggact
      481 ttttggagct ccgccactgt ccgaggggta cgtggctttg ttgggggacg agagacagag
      541 acacttcccg cccccgtctg aatttttgct ttcggtttta cgccgaaacc gcgccgcgcg
      601 tcttgtctgc tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc
      661 tgaaaattag ctcgacaaag ttaagtaata gtccctctct ccaagctcac ttacaggcgg
      721 ccgccatgct ctgttgcatg aggcggacta agcaggtgga aaaaaacgat gaagaccaaa
      781 aaattatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
      841 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
      901 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
      961 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     1021 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     1081 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     1141 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     1201 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     1261 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     1321 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1381 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     1441 tggtcctgct ggagttcgtg accgccgccg ggatcactct cgggtcatca tcaagtgctg
     1501 ttaatactct tgttgagaga tggaatgaaa aatacgcaca agcatacccc aatgtctccg
     1561 ctaagataga ttatgacgac gaaatgggaa gtagcagtag cgcaaaggtt ccacgcaatc
     1621 aggactggtt gtgagaattc gagcatctta ccgccattta ttcccatatt tgttctgttt
     1681 ttcttgattt gggtatacat ttaaatgtta ataaaacaaa atggtggggc aatcatttac
     1741 attttatggg atatgtaatt actagttcag gtgtattgcc acaagacaaa catgttaaga
     1801 aactttcccg ttatttacgc tctgttcctg ttaatcaacc tctggattac aaaatttgtg
     1861 aaagattgac tgatattctt aactatgttg ctccttttac gctgtgtgga tatgctgctt
     1921 taatgcctct gtatcatgct attgcttccc gtacggcttt cgttttctcc tccttgtata
     1981 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtccgtcaa cgtggcgtgg
     2041 tgtgctctgt gtttgctgac gcaaccccca ctggctgggg cattgccacc acctgtcaac
     2101 tcctttctgg gactttcgct ttccccctcc cgatcgccac ggcagaactc atcgccgcct
     2161 gccttgcccg ctgctggaca ggggctaggt tgctgggcac tgataattcc gtggtgttgt
     2221 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccaactgg atcctgcgcg
     2281 ggacgtcctt ctgctacgtc ccttcggctc tcaatccagc ggacctccct tcccgaggcc
     2341 ttctgccggt tctgcggcct ctcccgcgtc ttcgctttcg gcctccgacg agtcggatct
     2401 ccctttgggc cgcctccccg cctgtttcgc ctcggcgtcc ggtccgtgtt gcttggtcgt
     2461 cacctgtgca gaattgcgaa ccatggattc caccgtgaac tttgtctcct ggcatgcaaa
     2521 tcgtcaactt ggcatgccaa gaataattcg gatccaagct taggcctgct cgctttcttg
     2581 ctgtcccatt tctattaaag gttcctttgt tccctaagtc caactactaa actgggggat
     2641 attatgaagg gccttgagca tctggattct gcctagcgct aagcttaaca cgagccatag
     2701 atagaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt
     2761 aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaat acataactga
     2821 gaatagagaa gttcagatca aggttaggaa cagagagaca gcagaatatg ggccaaacag
     2881 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagttg gaacagcaga
     2941 atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac
     3001 agatggtccc cagatgcggt cccgccctca gcagtttcta gagaaccatc agatgtttcc
     3061 agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc
     3121 ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct
     3181 cactcggcgc gccagtcctc cgatagactg cgtcgcccgg gtacccgtgt tctcaataaa
     3241 ccctcttgca gttgcatccg actcgtggtc tcgctgttcc ttgggagggt ctcctctgag
     3301 tgattgactg cccacctcgg gggtctttca ttctcgagca gcttggcgta atcatggtca
     3361 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     3421 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     3481 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     3541 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgctccctc gctcactgac
     3601 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3661 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3721 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3781 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3841 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3901 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3961 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4021 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4081 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4141 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4201 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4261 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4321 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4381 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4441 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4501 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4561 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4621 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4681 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4741 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4801 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     4861 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4921 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4981 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5041 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5101 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5161 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5221 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5281 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5341 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5401 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5461 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     5521 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     5581 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     5641 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     5701 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     5761 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat
     5821 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     5881 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     5941 tcccagtcac gacgttgtaa aacgacggcc agtgaattag tactctagct taagtaacgc
     6001 cattttgcaa ggcatggaaa atacataact gagaatagag aagttcagat caaggttagg
     6061 aacagagaga cagcagaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
     6121 ggctcagggc caagaacagt tggaacagca gaatatgggc caaacaggat atctgtggta
     6181 agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg gtcccgccct
     6241 cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc
     6301 tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct
     6361 c
//
