LOCUS       sequence-344448-        7132 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses membrane-associated (palmitoylated) EGFP and extracellular
            and transmembrane domains of murine CD19.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7132)
  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 7132)
  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-tCD19-mGFP
FEATURES             Location/Qualifiers
     source          1..7132
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1669..1671,1672..1674,1675..2388)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1669..1671,1672..1674,1675..2385)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1714..1735)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1975..1994)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2322..2343
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2587..2990
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             3496..4086
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4111..4127)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4111..4127)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4124..4146)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4135..4151
                     /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(4159..4165,4166..4183,4184..4189))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4204..4225
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4342..4359)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4513..5101)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4593..4612)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5272..6063,6064..6132))
                     /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     5895..5914
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6133..6237)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6305..6323
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6361..6383)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6483..6502
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6696..6718
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6710..6727
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6711..6727
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 agcccacaac ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc
       61 gtattcccaa taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg
      121 agggtctcct cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc
      181 gagatttgga gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca
      241 gcggtcgttt cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc
      301 gcctgcgtct gtactagttg gctaactaga tctgtatctg gcggtcccgc ggaagaactg
      361 acgagttcgt attcccggcc gcagcccctg ggagacgtcc cagcggcctc gggggcccgt
      421 tttgtggccc attctgtatc agttaaccta cccgagtcgg actttttgga gctccgccac
      481 tgtccgaggg gtacgtggct ttgttggggg acgagagaca gagacacttc ccgcccccgt
      541 ctgaattttt gctttcggtt ttacgccgaa accgcgccgc gcgtcttgtc tgctgcagca
      601 tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt gtctgaaaat tagctcgaca
      661 aagttaagta atagtccctc tctccaagct cacttacagg cggccgccat gccatctcct
      721 ctccctgtct ccttcctcct ctttcttacc ttagtaggag gcaggcccca gaagtcctta
      781 ctggtggagg tagaagaggg aggcaatgtt gtgctgccat gcctcccgga ctcctcacct
      841 gtctcttctg agaagctggc ttggtatcga ggtaaccagt caacaccctt cctggagctg
      901 agccccgggt cccctggcct gggattgcac gtggggtccc tgggcatctt gctagtgatt
      961 gtcaatgtct cagaccatat ggggggcttc tacctgtgcc agaagaggcc ccctttcaag
     1021 gacatctggc agcctgcctg gacagtgaac gtggaggata gtggggagat gttccggtgg
     1081 aatgcttcag acgtcaggga cctggactgt gacctaagga acaggtcctc tgggagccac
     1141 aggtccactt ctggttccca gctgtatgtg tgggctaaag accatcctaa ggtctgggga
     1201 acaaagcctg tatgtgcccc tcgggggagc agtttgaatc agagtctaat caaccaagat
     1261 ctcactgtgg cacccggctc cacactttgg ctgtcctgtg gggtaccccc tgtcccagtg
     1321 gccaaaggct ccatctcctg gacccatgtg catcctagga gacctaatgt ttcactactg
     1381 agcctaagcc ttgggggaga gcacccggtc agagagatgt gggtttgggg gtctcttctg
     1441 cttctgcccc aagccacagc tttagatgaa ggcacctatt attgtctccg aggaaacctg
     1501 accatcgaga ggcacgtgaa ggtcattgca aggtcagtgt ggctctggct gttgagaact
     1561 ggtggatgga tagtcccagt ggtgacttta gtatatgtca tcttctgtat ggtttctctg
     1621 gtggcttttc tctattgtca aagagccttt atcctgagaa ggaaaaggat ggtgagcaag
     1681 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     1741 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     1801 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     1861 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     1921 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     1981 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     2041 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     2101 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     2161 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     2221 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     2281 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     2341 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaga attcgagcat
     2401 cttaccgcca tttattccca tatttgttct gtttttcttg atttgggtat acatttaaat
     2461 gttaataaaa caaaatggtg gggcaatcat ttacatttta tgggatatgt aattactagt
     2521 tcaggtgtat tgccacaaga caaacatgtt aagaaacttt cccgttattt acgctctgtt
     2581 cctgttaatc aacctctgga ttacaaaatt tgtgaaagat tgactgatat tcttaactat
     2641 gttgctcctt ttacgctgtg tggatatgct gctttaatgc ctctgtatca tgctattgct
     2701 tcccgtacgg ctttcgtttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     2761 gagttgtggc ccgttgtccg tcaacgtggc gtggtgtgct ctgtgtttgc tgacgcaacc
     2821 cccactggct ggggcattgc caccacctgt caactccttt ctgggacttt cgctttcccc
     2881 ctcccgatcg ccacggcaga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     2941 aggttgctgg gcactgataa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
     3001 ctgctcgcct gtgttgccaa ctggatcctg cgcgggacgt ccttctgcta cgtcccttcg
     3061 gctctcaatc cagcggacct cccttcccga ggccttctgc cggttctgcg gcctctcccg
     3121 cgtcttcgct ttcggcctcc gacgagtcgg atctcccttt gggccgcctc cccgcctgtt
     3181 tcgcctcggc gtccggtccg tgttgcttgg tcgtcacctg tgcagaattg cgaaccatgg
     3241 attccaccgt gaactttgtc tcctggcatg caaatcgtca acttggcatg ccaagaataa
     3301 ttcggatcca agcttaggcc tgctcgcttt cttgctgtcc catttctatt aaaggttcct
     3361 ttgttcccta agtccaacta ctaaactggg ggatattatg aagggccttg agcatctgga
     3421 ttctgcctag cgctaagctt aacacgagcc atagatagaa taaaagattt tatttagtct
     3481 ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac
     3541 gccattttgc aaggcatgga aaatacataa ctgagaatag agaagttcag atcaaggtta
     3601 ggaacagaga gacagcagaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
     3661 ccggctcagg gccaagaaca gttggaacag cagaatatgg gccaaacagg atatctgtgg
     3721 taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtcccgcc
     3781 ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac
     3841 cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg
     3901 ctccccgagc tcaataaaag agcccacaac ccctcactcg gcgcgccagt cctccgatag
     3961 actgcgtcgc ccgggtaccc gtgttctcaa taaaccctct tgcagttgca tccgactcgt
     4021 ggtctcgctg ttccttggga gggtctcctc tgagtgattg actgcccacc tcgggggtct
     4081 ttcattctcg agcagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4141 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     4201 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4261 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4321 tattgggcgc tcttccgctc cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4381 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4441 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4501 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4561 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4621 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4681 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4741 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4801 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4861 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4921 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4981 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5041 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5101 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5161 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5221 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5281 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5341 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5401 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5461 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5521 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5581 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5641 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5701 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5761 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5821 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5881 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5941 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6001 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6061 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6121 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6181 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6241 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     6301 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     6361 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     6421 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     6481 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     6541 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     6601 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     6661 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     6721 ggccagtgaa ttagtactct agcttaagta acgccatttt gcaaggcatg gaaaatacat
     6781 aactgagaat agagaagttc agatcaaggt taggaacaga gagacagcag aatatgggcc
     6841 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagttggaac
     6901 agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     6961 aagaacagat ggtccccaga tgcggtcccg ccctcagcag tttctagaga accatcagat
     7021 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     7081 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa ag
//
