LOCUS       sequence-344453-        6199 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stably expresses the extracellular and transmembrane domain of 
            murine CD40L.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6199)
  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 6199)
  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-CD40L
FEATURES             Location/Qualifiers
     source          1..6199
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1032..1041
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1983..2386
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             2892..3482
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3507..3523)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3507..3523)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3520..3542)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3531..3547
                     /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(3555..3561,3562..3579,3580..3585))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3600..3621
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3738..3755)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3909..4497)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3989..4008)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4668..5459,5460..5528))
                     /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     5291..5310
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5529..5633)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5701..5719
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5757..5779)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5879..5898
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6092..6114
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6106..6123
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6107..6123
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tcaaggttag gaacagagag acagcagaat atgggccaaa caggatatct gtggtaagca
       61 gttcctgccc cggctcaggg ccaagaacag ttggaacagc agaatatggg ccaaacagga
      121 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc
      181 ggtcccgccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc
      241 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
      301 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc
      361 ctccgataga ctgcgtcgcc cgggtacccg tattcccaat aaagcctctt gctgtttgca
      421 tccgaatcgt ggactcgctg atccttggga gggtctcctc agattgattg actgcccacc
      481 tcgggggtct ttcatttgga ggttccaccg agatttggag acccctgccc agggaccacc
      541 gacccccccg ccgggaggta agctggccag cggtcgtttc gtgtctgtct ctgtctttgt
      601 gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg tactagttgg ctaactagat
      661 ctgtatctgg cggtcccgcg gaagaactga cgagttcgta ttcccggccg cagcccctgg
      721 gagacgtccc agcggcctcg ggggcccgtt ttgtggccca ttctgtatca gttaacctac
      781 ccgagtcgga ctttttggag ctccgccact gtccgagggg tacgtggctt tgttggggga
      841 cgagagacag agacacttcc cgcccccgtc tgaatttttg ctttcggttt tacgccgaaa
      901 ccgcgccgcg cgtcttgtct gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt
      961 tctgtatttg tctgaaaatt agctcgacaa agttaagtaa tagtccctct ctccaagctc
     1021 acttacaggc ggccgccatg gctacaggcc tgcccgccag catgaagatc tttatgtacc
     1081 tgctgaccgt gttcctgatc acccagatga tcggcagcgt gctgttcgcc gtgtacctgc
     1141 acagacggct ggacaaggtg gaagaggaag tgaacctgca cgaggacttc gtgttcatca
     1201 agaaactgaa gcggtgcaac aagggcgagg gcagcctgag cctgctgaac tgcgaggaaa
     1261 tgagaaggca gttcgaggac ctcgtgaagg acatcaccct gaacaaagag gaaaagaaag
     1321 aaaactcctt cgagatgcag aggggcgacg aggaccctca gatcgctgct cacgtggtgt
     1381 ccgaggccaa cagcaacgcc gcttctgtgc tgcagtgggc caagaaaggc tactacacca
     1441 tgaagtccaa cctcgtgatg ctggaaaacg gcaagcagct gacagtgaag cgcgagggcc
     1501 tgtactatgt gtacacccaa gtgacattct gcagcaacag agagcccagc agccagaggc
     1561 ccttcatcgt gggactgtgg ctgaagccta gcagcggcag cgagagaatc ctgctgaagg
     1621 ccgccaacac ccacagcagc tctcagctgt gcgagcagca gagcgtgcac ctgggcggag
     1681 tgttcgagct gcaagctggc gcctccgtgt tcgtgaacgt gacagaggcc agccaagtga
     1741 tccacagagt gggcttcagc agctttggac tgctgaaact gtgagaattc gagcatctta
     1801 ccgccattta ttcccatatt tgttctgttt ttcttgattt gggtatacat ttaaatgtta
     1861 ataaaacaaa atggtggggc aatcatttac attttatggg atatgtaatt actagttcag
     1921 gtgtattgcc acaagacaaa catgttaaga aactttcccg ttatttacgc tctgttcctg
     1981 ttaatcaacc tctggattac aaaatttgtg aaagattgac tgatattctt aactatgttg
     2041 ctccttttac gctgtgtgga tatgctgctt taatgcctct gtatcatgct attgcttccc
     2101 gtacggcttt cgttttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     2161 tgtggcccgt tgtccgtcaa cgtggcgtgg tgtgctctgt gtttgctgac gcaaccccca
     2221 ctggctgggg cattgccacc acctgtcaac tcctttctgg gactttcgct ttccccctcc
     2281 cgatcgccac ggcagaactc atcgccgcct gccttgcccg ctgctggaca ggggctaggt
     2341 tgctgggcac tgataattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc
     2401 tcgcctgtgt tgccaactgg atcctgcgcg ggacgtcctt ctgctacgtc ccttcggctc
     2461 tcaatccagc ggacctccct tcccgaggcc ttctgccggt tctgcggcct ctcccgcgtc
     2521 ttcgctttcg gcctccgacg agtcggatct ccctttgggc cgcctccccg cctgtttcgc
     2581 ctcggcgtcc ggtccgtgtt gcttggtcgt cacctgtgca gaattgcgaa ccatggattc
     2641 caccgtgaac tttgtctcct ggcatgcaaa tcgtcaactt ggcatgccaa gaataattcg
     2701 gatccaagct taggcctgct cgctttcttg ctgtcccatt tctattaaag gttcctttgt
     2761 tccctaagtc caactactaa actgggggat attatgaagg gccttgagca tctggattct
     2821 gcctagcgct aagcttaaca cgagccatag atagaataaa agattttatt tagtctccag
     2881 aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca
     2941 ttttgcaagg catggaaaat acataactga gaatagagaa gttcagatca aggttaggaa
     3001 cagagagaca gcagaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
     3061 ctcagggcca agaacagttg gaacagcaga atatgggcca aacaggatat ctgtggtaag
     3121 cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt cccgccctca
     3181 gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg
     3241 tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc
     3301 ccgagctcaa taaaagagcc cacaacccct cactcggcgc gccagtcctc cgatagactg
     3361 cgtcgcccgg gtacccgtgt tctcaataaa ccctcttgca gttgcatccg actcgtggtc
     3421 tcgctgttcc ttgggagggt ctcctctgag tgattgactg cccacctcgg gggtctttca
     3481 ttctcgagca gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     3541 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     3601 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     3661 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     3721 gggcgctctt ccgctccctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3781 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3841 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3901 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3961 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4021 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4081 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4141 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4201 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4261 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4321 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4381 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4441 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4501 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4561 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4621 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4681 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4741 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4801 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4861 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4921 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4981 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5041 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5101 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5161 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5221 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5281 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5341 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5401 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5461 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5521 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5581 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5641 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     5701 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     5761 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     5821 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     5881 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     5941 gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa
     6001 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     6061 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     6121 agtgaattag tactctagct taagtaacgc cattttgcaa ggcatggaaa atacataact
     6181 gagaatagag aagttcaga
//
