LOCUS       sequence-344458-        5935 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses extracellular domain of human IL-2 (aa 21-153) (mutant H9 
            containing the mutations L80F / R81D / L85V / I 86V / I92F) 
            developed by the Garcia Lab.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5935)
  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 5935)
  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-super-IL-2
FEATURES             Location/Qualifiers
     source          1..5935
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     173..191
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(229..251)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     351..370
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     564..586
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     578..595
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     579..595
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      1703..1712
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2390..2793
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             3299..3889
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3914..3930)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3914..3930)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3927..3949)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3938..3954
                     /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(3962..3968,3969..3986,3987..3992))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4007..4028
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4145..4162)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4316..4904)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4396..4415)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5075..5866,5867..5935))
                     /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     5698..5717
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
      181 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
      241 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
      301 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
      361 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
      421 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
      481 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
      541 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
      601 agtactctag cttaagtaac gccattttgc aaggcatgga aaatacataa ctgagaatag
      661 agaagttcag atcaaggtta ggaacagaga gacagcagaa tatgggccaa acaggatatc
      721 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gttggaacag cagaatatgg
      781 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
      841 tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt ttccagggtg
      901 ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg
      961 cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg
     1021 gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa taaagcctct
     1081 tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct cagattgatt
     1141 gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga gacccctgcc
     1201 cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt cgtgtctgtc
     1261 tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct gtactagttg
     1321 gctaactaga tctgtatctg gcggtcccgc ggaagaactg acgagttcgt attcccggcc
     1381 gcagcccctg ggagacgtcc cagcggcctc gggggcccgt tttgtggccc attctgtatc
     1441 agttaaccta cccgagtcgg actttttgga gctccgccac tgtccgaggg gtacgtggct
     1501 ttgttggggg acgagagaca gagacacttc ccgcccccgt ctgaattttt gctttcggtt
     1561 ttacgccgaa accgcgccgc gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt
     1621 ctgactgtgt ttctgtattt gtctgaaaat tagctcgaca aagttaagta atagtccctc
     1681 tctccaagct cacttacagg cggccgccat ggctagccct ctgacaagat tcctgagcct
     1741 gaacctgctg ctgctgggcg agtctatcat cctcggatct ggcgaggccg ctcctaccag
     1801 cagctctacc aagaaaaccc agctccagct cgagcacctc ctgctggacc tgcagatgat
     1861 cctgaacggc atcaacaact acaagaaccc caagctgacc cggatgctga ccttcaagtt
     1921 ctacatgccc aagaaggcca ccgagctgaa gcacctccag tgcctggaag aggaactgaa
     1981 gcccctggaa gaagtgctga acctggctca gagcaagaac ttccacttcg accctcgcga
     2041 cgtggtgtcc aacatcaacg tgttcgtgct ggaactgaaa ggcagcgaga caaccttcat
     2101 gtgcgagtac gccgacgaga cagctacaat cgtggaattc ctgaaccggt ggatcacctt
     2161 ctgccagagc atcatcagca ccctgacctg agaattcgag catcttaccg ccatttattc
     2221 ccatatttgt tctgtttttc ttgatttggg tatacattta aatgttaata aaacaaaatg
     2281 gtggggcaat catttacatt ttatgggata tgtaattact agttcaggtg tattgccaca
     2341 agacaaacat gttaagaaac tttcccgtta tttacgctct gttcctgtta atcaacctct
     2401 ggattacaaa atttgtgaaa gattgactga tattcttaac tatgttgctc cttttacgct
     2461 gtgtggatat gctgctttaa tgcctctgta tcatgctatt gcttcccgta cggctttcgt
     2521 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     2581 ccgtcaacgt ggcgtggtgt gctctgtgtt tgctgacgca acccccactg gctggggcat
     2641 tgccaccacc tgtcaactcc tttctgggac tttcgctttc cccctcccga tcgccacggc
     2701 agaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctaggttgc tgggcactga
     2761 taattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     2821 caactggatc ctgcgcggga cgtccttctg ctacgtccct tcggctctca atccagcgga
     2881 cctcccttcc cgaggccttc tgccggttct gcggcctctc ccgcgtcttc gctttcggcc
     2941 tccgacgagt cggatctccc tttgggccgc ctccccgcct gtttcgcctc ggcgtccggt
     3001 ccgtgttgct tggtcgtcac ctgtgcagaa ttgcgaacca tggattccac cgtgaacttt
     3061 gtctcctggc atgcaaatcg tcaacttggc atgccaagaa taattcggat ccaagcttag
     3121 gcctgctcgc tttcttgctg tcccatttct attaaaggtt cctttgttcc ctaagtccaa
     3181 ctactaaact gggggatatt atgaagggcc ttgagcatct ggattctgcc tagcgctaag
     3241 cttaacacga gccatagata gaataaaaga ttttatttag tctccagaaa aaggggggaa
     3301 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
     3361 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
     3421 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
     3481 acagttggaa cagcagaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
     3541 ggctcagggc caagaacaga tggtccccag atgcggtccc gccctcagca gtttctagag
     3601 aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa
     3661 ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa
     3721 aagagcccac aacccctcac tcggcgcgcc agtcctccga tagactgcgt cgcccgggta
     3781 cccgtgttct caataaaccc tcttgcagtt gcatccgact cgtggtctcg ctgttccttg
     3841 ggagggtctc ctctgagtga ttgactgccc acctcggggg tctttcattc tcgagcagct
     3901 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3961 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     4021 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     4081 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     4141 ctccctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     4201 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4261 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4321 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4381 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4441 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4501 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4561 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4621 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4681 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4741 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4801 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4861 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4921 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4981 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     5041 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     5101 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     5161 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     5221 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     5281 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     5341 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     5401 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     5461 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5521 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5581 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5641 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5701 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5761 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5821 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5881 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcat
//
