LOCUS       sequence-344460-        7558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a constitutively activated murine IL-12 receptor tagged 
            with extracellular myc and flag tags.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7558)
  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 7558)
  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-mIL15-IL12rb1-sushi-il12rb2
FEATURES             Location/Qualifiers
     source          1..7558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      737..746
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             827..850
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1745..1801
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1880..1909
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    3047..3450
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             3956..4546
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4571..4587)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4571..4587)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4584..4606)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4595..4611
                     /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(4619..4625,4626..4643,4644..4649))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4664..4685
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4802..4819)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4973..5561)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5053..5072)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5732..6523,6524..6592))
                     /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     6355..6374
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6593..6697)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6765..6783
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6821..6843)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6943..6962
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7156..7178
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7170..7187
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7171..7187
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca ctcggcgcgc
       61 cagtcctccg atagactgcg tcgcccgggt acccgtattc ccaataaagc ctcttgctgt
      121 ttgcatccga atcgtggact cgctgatcct tgggagggtc tcctcagatt gattgactgc
      181 ccacctcggg ggtctttcat ttggaggttc caccgagatt tggagacccc tgcccaggga
      241 ccaccgaccc ccccgccggg aggtaagctg gccagcggtc gtttcgtgtc tgtctctgtc
      301 tttgtgcgtg tttgtgccgg catctaatgt ttgcgcctgc gtctgtacta gttggctaac
      361 tagatctgta tctggcggtc ccgcggaaga actgacgagt tcgtattccc ggccgcagcc
      421 cctgggagac gtcccagcgg cctcgggggc ccgttttgtg gcccattctg tatcagttaa
      481 cctacccgag tcggactttt tggagctccg ccactgtccg aggggtacgt ggctttgttg
      541 ggggacgaga gacagagaca cttcccgccc ccgtctgaat ttttgctttc ggttttacgc
      601 cgaaaccgcg ccgcgcgtct tgtctgctgc agcatcgttc tgtgttgtct ctgtctgact
      661 gtgtttctgt atttgtctga aaattagctc gacaaagtta agtaatagtc cctctctcca
      721 agctcactta caggcggccg ccatggctag ccctctgaca agattcctga gcctgaacct
      781 gctgctgctg ggcgagtcta tcatcctcgg atctggcgag gccggcgact acaaggacga
      841 tgacgacaaa ggcaactggg tcaacgtgat cagcagcctg aagaagatcg aggacctgat
      901 ccagagcatg cacatcgacg ccacactgta caccgagagc gacgtgcacc ctagctgtaa
      961 agtgaccgct atgaagtgct tcctgctcga gctgcaagtg atcagcctgg aatctggcga
     1021 cgccagcatc cacgacaccg tggaaaacct gatcatcctg gccaacaaca gcctgagcag
     1081 caacggcaac gtgaccgagt ctggctgcaa agagtgcgag gaactggaag agaagaacat
     1141 caaagagttc ctccagagct tcgtgcacat cgtgtccatg ttcatcaaca ccagcggcag
     1201 cgaggtgcag atcagcagac tgtccatcat cttcgccagc ctgggcagct tcgcttctgt
     1261 gctgcttgtt ggctccctgg gctacatcgg cctgaacaga gctgcctggc atctgtgtcc
     1321 tccactgcct acaccttgtg gcagcaccgc tgtggaattc cctggctctc agggaaagca
     1381 ggcctggcag tggtgtaacc ctgaggactt tcccgaggtg ctgtacccca gagatgccct
     1441 ggtggttgag atgcctggcg atagaggcga cggaacagag tctcctcagg ccgctcctga
     1501 atgtgccctg gacacaagaa ggcccctgga aacccagaga cagagacagg tgcaggccct
     1561 gtctgaggct agaaggctgg gactcgccag agaggattgc cctagaggcg atctggctca
     1621 cgtgacactg cctttgctgc ttggcggagt tacacagggc gctagcgtgc tggacgacct
     1681 ttggagaacc cacaagacag ccgagcctgg acctcctaca ctgggacaag aagctggctc
     1741 tggcgccacc aactttagcc tgctgaaaca ggcaggcgac gtggaagaaa accccggacc
     1801 tatgtgcaga gccatcagcc tgagaaggct gctgctcctg ctgcttcagc tgtctcagct
     1861 gctggctgtg actcagggcg agcagaagct gatctccgag gaagatctgg gcatcacctg
     1921 tccacctcca atgagcgtgg aacacgccga catctgggtc aagagctaca gcctgtacag
     1981 cagagagcgg tacatctgca acagcggctt caagagaaag gccggcacct ctagcctgac
     2041 cgagtgcgtg ctgaacaagg ccacaaatgt ggcccactgg accacaccta gcctgaagtg
     2101 catcagagat ggcagccctc agggcaaagc caactggaag gcctttgtga tcagctccat
     2161 ctgtatcgcc atcatcaccg tgggcacctt cagcatcaga tacttcagac agaaggcttt
     2221 caccctgctg agcaccctga agcctcagtg gtacagcagg acaatccccg atcctgccaa
     2281 cagcacctgg gttaagaagt accccatcct cgaagaaaag atccagctgc ctaccgacaa
     2341 cctgctgatg gcctggccta caccagagga acctgagcct ctgatcattc acgaagtgct
     2401 ctaccacatg atccccgtcg tgcggcagcc ctactacttt aagagaggcc agggcttcca
     2461 gggctactcc acatctaagc aggacgccat gtatatcgct aaccctcagg ccaccggcac
     2521 actgaccgct gaaactagac agctggtcaa cctgtacaag gtgctggaaa gcagagatcc
     2581 cgacagcaag ctggctaacc tgacaagccc actgaccgtg acacccgtga actacctgcc
     2641 tagccacgag ggctacctgc caagcaacat cgaggatctg agccctcacg aggccgatcc
     2701 aacagacagc ttcgacctgg aacaccagca catctccctg agcatcttcg cttcaagcag
     2761 cctgcggcca ctgatctttg gcggcgaaag actgaccctg gacagactga agatgggcta
     2821 cgacagcctg atgtccaacg aggcttgaga attcgagcat cttaccgcca tttattccca
     2881 tatttgttct gtttttcttg atttgggtat acatttaaat gttaataaaa caaaatggtg
     2941 gggcaatcat ttacatttta tgggatatgt aattactagt tcaggtgtat tgccacaaga
     3001 caaacatgtt aagaaacttt cccgttattt acgctctgtt cctgttaatc aacctctgga
     3061 ttacaaaatt tgtgaaagat tgactgatat tcttaactat gttgctcctt ttacgctgtg
     3121 tggatatgct gctttaatgc ctctgtatca tgctattgct tcccgtacgg ctttcgtttt
     3181 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtccg
     3241 tcaacgtggc gtggtgtgct ctgtgtttgc tgacgcaacc cccactggct ggggcattgc
     3301 caccacctgt caactccttt ctgggacttt cgctttcccc ctcccgatcg ccacggcaga
     3361 actcatcgcc gcctgccttg cccgctgctg gacaggggct aggttgctgg gcactgataa
     3421 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccaa
     3481 ctggatcctg cgcgggacgt ccttctgcta cgtcccttcg gctctcaatc cagcggacct
     3541 cccttcccga ggccttctgc cggttctgcg gcctctcccg cgtcttcgct ttcggcctcc
     3601 gacgagtcgg atctcccttt gggccgcctc cccgcctgtt tcgcctcggc gtccggtccg
     3661 tgttgcttgg tcgtcacctg tgcagaattg cgaaccatgg attccaccgt gaactttgtc
     3721 tcctggcatg caaatcgtca acttggcatg ccaagaataa ttcggatcca agcttaggcc
     3781 tgctcgcttt cttgctgtcc catttctatt aaaggttcct ttgttcccta agtccaacta
     3841 ctaaactggg ggatattatg aagggccttg agcatctgga ttctgcctag cgctaagctt
     3901 aacacgagcc atagatagaa taaaagattt tatttagtct ccagaaaaag gggggaatga
     3961 aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga
     4021 aaatacataa ctgagaatag agaagttcag atcaaggtta ggaacagaga gacagcagaa
     4081 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
     4141 gttggaacag cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc
     4201 tcagggccaa gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac
     4261 catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta
     4321 accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag
     4381 agcccacaac ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc
     4441 gtgttctcaa taaaccctct tgcagttgca tccgactcgt ggtctcgctg ttccttggga
     4501 gggtctcctc tgagtgattg actgcccacc tcgggggtct ttcattctcg agcagcttgg
     4561 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     4621 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     4681 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4741 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctc
     4801 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4861 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4921 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4981 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5041 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5101 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5161 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5221 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5281 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5341 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5401 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5461 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5521 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5581 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5641 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5701 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5761 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5821 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5881 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5941 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6001 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6061 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6121 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6181 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6241 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6301 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6361 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6421 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6481 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6541 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6601 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6661 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6721 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     6781 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     6841 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     6901 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     6961 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     7021 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     7081 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     7141 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttagtactct
     7201 agcttaagta acgccatttt gcaaggcatg gaaaatacat aactgagaat agagaagttc
     7261 agatcaaggt taggaacaga gagacagcag aatatgggcc aaacaggata tctgtggtaa
     7321 gcagttcctg ccccggctca gggccaagaa cagttggaac agcagaatat gggccaaaca
     7381 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga
     7441 tgcggtcccg ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg
     7501 acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctg
//
