LOCUS       sequence-344451-        6361 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses palmitoylated GFP with a C terminal fusion of the LLO190 
            epitope and the murine 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-mPMEL
FEATURES             Location/Qualifiers
     source          1..6361
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(654..656,657..659,660..1172)
                     /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(699..720)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(960..979)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1307..1328
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1701..2104
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             2610..3200
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3225..3241)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3225..3241)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3238..3260)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3249..3265
                     /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(3273..3279,3280..3297,3298..3303))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3318..3339
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3456..3473)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3627..4215)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3707..3726)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4386..5177,5178..5246))
                     /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     5009..5028
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5247..5351)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5419..5437
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5475..5497)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5597..5616
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5810..5832
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5824..5841
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5825..5841
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ttgggagggt ctcctcagat tgattgactg cccacctcgg gggtctttca tttggaggtt
       61 ccaccgagat ttggagaccc ctgcccaggg accaccgacc cccccgccgg gaggtaagct
      121 ggccagcggt cgtttcgtgt ctgtctctgt ctttgtgcgt gtttgtgccg gcatctaatg
      181 tttgcgcctg cgtctgtact agttggctaa ctagatctgt atctggcggt cccgcggaag
      241 aactgacgag ttcgtattcc cggccgcagc ccctgggaga cgtcccagcg gcctcggggg
      301 cccgttttgt ggcccattct gtatcagtta acctacccga gtcggacttt ttggagctcc
      361 gccactgtcc gaggggtacg tggctttgtt gggggacgag agacagagac acttcccgcc
      421 cccgtctgaa tttttgcttt cggttttacg ccgaaaccgc gccgcgcgtc ttgtctgctg
      481 cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg aaaattagct
      541 cgacaaagtt aagtaatagt ccctctctcc aagctcactt acaggcggcc gccatgctct
      601 gttgcatgag gcggactaag caggtggaaa aaaacgatga agaccaaaaa attatggtga
      661 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
      721 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
      781 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
      841 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
      901 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
      961 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     1021 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     1081 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     1141 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     1201 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     1261 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     1321 agttcgtgac cgccgccggg atcactctcg ggtcatcatc aagtgctgtt aatactcttg
     1381 ttgagagatg gaatgaaaaa tacgcacaag cataccccaa tgtctccgct aagatagatt
     1441 atgacgacga aatgggaagt agcagtagcg ccgaaggtag tagaaaccaa gattggttgt
     1501 gagaattcga gcatcttacc gccatttatt cccatatttg ttctgttttt cttgatttgg
     1561 gtatacattt aaatgttaat aaaacaaaat ggtggggcaa tcatttacat tttatgggat
     1621 atgtaattac tagttcaggt gtattgccac aagacaaaca tgttaagaaa ctttcccgtt
     1681 atttacgctc tgttcctgtt aatcaacctc tggattacaa aatttgtgaa agattgactg
     1741 atattcttaa ctatgttgct ccttttacgc tgtgtggata tgctgcttta atgcctctgt
     1801 atcatgctat tgcttcccgt acggctttcg ttttctcctc cttgtataaa tcctggttgc
     1861 tgtctcttta tgaggagttg tggcccgttg tccgtcaacg tggcgtggtg tgctctgtgt
     1921 ttgctgacgc aacccccact ggctggggca ttgccaccac ctgtcaactc ctttctggga
     1981 ctttcgcttt ccccctcccg atcgccacgg cagaactcat cgccgcctgc cttgcccgct
     2041 gctggacagg ggctaggttg ctgggcactg ataattccgt ggtgttgtcg gggaagctga
     2101 cgtcctttcc atggctgctc gcctgtgttg ccaactggat cctgcgcggg acgtccttct
     2161 gctacgtccc ttcggctctc aatccagcgg acctcccttc ccgaggcctt ctgccggttc
     2221 tgcggcctct cccgcgtctt cgctttcggc ctccgacgag tcggatctcc ctttgggccg
     2281 cctccccgcc tgtttcgcct cggcgtccgg tccgtgttgc ttggtcgtca cctgtgcaga
     2341 attgcgaacc atggattcca ccgtgaactt tgtctcctgg catgcaaatc gtcaacttgg
     2401 catgccaaga ataattcgga tccaagctta ggcctgctcg ctttcttgct gtcccatttc
     2461 tattaaaggt tcctttgttc cctaagtcca actactaaac tgggggatat tatgaagggc
     2521 cttgagcatc tggattctgc ctagcgctaa gcttaacacg agccatagat agaataaaag
     2581 attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag
     2641 ctagcttaag taacgccatt ttgcaaggca tggaaaatac ataactgaga atagagaagt
     2701 tcagatcaag gttaggaaca gagagacagc agaatatggg ccaaacagga tatctgtggt
     2761 aagcagttcc tgccccggct cagggccaag aacagttgga acagcagaat atgggccaaa
     2821 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca
     2881 gatgcggtcc cgccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa
     2941 ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg
     3001 ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca ctcggcgcgc
     3061 cagtcctccg atagactgcg tcgcccgggt acccgtgttc tcaataaacc ctcttgcagt
     3121 tgcatccgac tcgtggtctc gctgttcctt gggagggtct cctctgagtg attgactgcc
     3181 cacctcgggg gtctttcatt ctcgagcagc ttggcgtaat catggtcata gctgtttcct
     3241 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     3301 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     3361 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3421 agaggcggtt tgcgtattgg gcgctcttcc gctccctcgc tcactgactc gctgcgctcg
     3481 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3541 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3601 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3661 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3721 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3781 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3841 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3901 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3961 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4021 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4081 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4141 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4201 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4261 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4321 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4381 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4441 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4501 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4561 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4621 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4681 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4741 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4801 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4861 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4921 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4981 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5041 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5101 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5161 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5221 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5281 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5341 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     5401 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     5461 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     5521 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     5581 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     5641 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg
     5701 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     5761 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     5821 cgttgtaaaa cgacggccag tgaattagta ctctagctta agtaacgcca ttttgcaagg
     5881 catggaaaat acataactga gaatagagaa gttcagatca aggttaggaa cagagagaca
     5941 gcagaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     6001 agaacagttg gaacagcaga atatgggcca aacaggatat ctgtggtaag cagttcctgc
     6061 cccggctcag ggccaagaac agatggtccc cagatgcggt cccgccctca gcagtttcta
     6121 gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt
     6181 gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa
     6241 taaaagagcc cacaacccct cactcggcgc gccagtcctc cgatagactg cgtcgcccgg
     6301 gtacccgtat tcccaataaa gcctcttgct gtttgcatcc gaatcgtgga ctcgctgatc
     6361 c
//
