LOCUS       sequence-343844-        6019 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses secreted murine interferon beta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6019)
  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 6019)
  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-IFN-B
FEATURES             Location/Qualifiers
     source          1..6019
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    949..1352
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     regulatory      2114..2123
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3231..3248)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3231..3247)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3240..3262)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3456..3475)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3575..3597
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3635..3653)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3721..3825
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3826..3894,3895..4686)
                     /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     complement(4044..4063)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4857..5445
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5346..5365
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5599..5616
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5733..5754
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5769..5774,5775..5792,5793..5799)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5807..5823
                     /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)."
     primer_bind     5812..5834
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5831..5847
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5831..5847
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     LTR             join(5872..6019,1..443)
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 agtgaggggt tgtgggctct tttattgagc tcggggagca gaagcgcgcg aacagaagcg
       61 agaagcgaac tgattggtta gttcaaataa ggcacagggt catttcaggt ccttggggca
      121 ccctggaaac atctgatggt tctctagaaa ctgctgaggg cgggaccgca tctggggacc
      181 atctgttctt ggccctgagc cggggcagga actgcttacc acagatatcc tgtttggccc
      241 atattctgct gttccaactg ttcttggccc tgagccgggg caggaactgc ttaccacaga
      301 tatcctgttt ggcccatatt ctgctgtctc tctgttccta accttgatct gaacttctct
      361 attctcagtt atgtattttc catgccttgc aaaatggcgt tacttaagct agcttgccaa
      421 acctacaggt ggggtctttc attcccccct ttttctggag actaaataaa atcttttatt
      481 ctatctatgg ctcgtgttaa gcttagcgct aggcagaatc cagatgctca aggcccttca
      541 taatatcccc cagtttagta gttggactta gggaacaaag gaacctttaa tagaaatggg
      601 acagcaagaa agcgagcagg cctaagcttg gatccgaatt attcttggca tgccaagttg
      661 acgatttgca tgccaggaga caaagttcac ggtggaatcc atggttcgca attctgcaca
      721 ggtgacgacc aagcaacacg gaccggacgc cgaggcgaaa caggcgggga ggcggcccaa
      781 agggagatcc gactcgtcgg aggccgaaag cgaagacgcg ggagaggccg cagaaccggc
      841 agaaggcctc gggaagggag gtccgctgga ttgagagccg aagggacgta gcagaaggac
      901 gtcccgcgca ggatccagtt ggcaacacag gcgagcagcc atggaaagga cgtcagcttc
      961 cccgacaaca ccacggaatt atcagtgccc agcaacctag cccctgtcca gcagcgggca
     1021 aggcaggcgg cgatgagttc tgccgtggcg atcgggaggg ggaaagcgaa agtcccagaa
     1081 aggagttgac aggtggtggc aatgccccag ccagtggggg ttgcgtcagc aaacacagag
     1141 cacaccacgc cacgttgacg gacaacgggc cacaactcct cataaagaga cagcaaccag
     1201 gatttataca aggaggagaa aacgaaagcc gtacgggaag caatagcatg atacagaggc
     1261 attaaagcag catatccaca cagcgtaaaa ggagcaacat agttaagaat atcagtcaat
     1321 ctttcacaaa ttttgtaatc cagaggttga ttaacaggaa cagagcgtaa ataacgggaa
     1381 agtttcttaa catgtttgtc ttgtggcaat acacctgaac tagtaattac atatcccata
     1441 aaatgtaaat gattgcccca ccattttgtt ttattaacat ttaaatgtat acccaaatca
     1501 agaaaaacag aacaaatatg ggaataaatg gcggtaagat gctcgaattc tcagttctgg
     1561 aagttccggg tcagccgtct gatgatcagg aagtttctga agatctcggc ccgcacgacc
     1621 atccaggcgt agctgttgta cttcatcagc ttcaggtatc tctgcacccg ccagtagtaa
     1681 gacttcaggt gcagggctgt gctgctcatc tcccaggtca ggcgttcctc ttgcttctct
     1741 tccagcacgg ttttcagaaa cacggtctgc tggtgcagct cgtccagcag ccgcacaacg
     1801 attgtctcgt tccagccggt gctgctgaag ttgttccgaa acaccaggaa cacgttctgc
     1861 agcatctctt ggatggcgaa ggcggtgtag gacttctgca tcttctcggt catttccatg
     1921 gggatcttga agtcggctct gtaggtcagg ttgatcttgc cgttcagctg ttccagcagc
     1981 tcttggcact ttctgatgtt ggtcctctct tgcagctgga gctgcttgta gttgatggcc
     2041 tcgccagatc cgaggatgat agactcgccc agcagcagca ggttcaggct caggaatctg
     2101 gtcagagggc tagccatggc ggccgcctgt aagtgagctt ggagagaggg actattactt
     2161 aactttgtcg agctaatttt cagacaaata cagaaacaca gtcagacaga gacaacacag
     2221 aacgatgctg cagcagacaa gacgcgcggc gcggtttcgg cgtaaaaccg aaagcaaaaa
     2281 ttcagacggg ggcgggaagt gtctctgtct ctcgtccccc aacaaagcca cgtacccctc
     2341 ggacagtggc ggagctccaa aaagtccgac tcgggtaggt taactgatac agaatgggcc
     2401 acaaaacggg cccccgaggc cgctgggacg tctcccaggg gctgcggccg ggaatacgaa
     2461 ctcgtcagtt cttccgcggg accgccagat acagatctag ttagccaact agtacagacg
     2521 caggcgcaaa cattagatgc cggcacaaac acgcacaaag acagagacag acacgaaacg
     2581 accgctggcc agcttacctc ccggcggggg ggtcggtggt ccctgggcag gggtctccaa
     2641 atctcggtgg aacctccaaa tgaaagaccc ccgaggtggg cagtcaatca atctgaggag
     2701 accctcccaa ggatcagcga gtccacgatt cggatgcaaa cagcaagagg ctttattggg
     2761 aatacgggta cccgggcgac gcagtctatc ggaggactgg cgcgccgagt gaggggttgt
     2821 gggctctttt attgagctcg gggagcagaa gcgcgcgaac agaagcgaga agcgaactga
     2881 ttggttagtt caaataaggc acagggtcat ttcaggtcct tggggcaccc tggaaacatc
     2941 tgatggttct ctagaaactg ctgagggcgg gaccgcatct ggggaccatc tgttcttggc
     3001 cctgagccgg ggcaggaact gcttaccaca gatatcctgt ttggcccata ttctgctgtt
     3061 ccaactgttc ttggccctga gccggggcag gaactgctta ccacagatat cctgtttggc
     3121 ccatattctg ctgtctctct gttcctaacc ttgatctgaa cttctctatt ctcagttatg
     3181 tattttccat gccttgcaaa atggcgttac ttaagctaga gtactaattc actggccgtc
     3241 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     3301 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     3361 cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg
     3421 tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag
     3481 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     3541 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     3601 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     3661 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     3721 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3781 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     3841 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     3901 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     3961 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     4021 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     4081 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     4141 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     4201 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     4261 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     4321 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     4381 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     4441 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     4501 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     4561 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     4621 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     4681 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     4741 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     4801 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4861 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4921 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4981 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     5041 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5101 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5161 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5221 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5281 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5341 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5401 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5461 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     5521 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     5581 agccgaacga ccgagcgcag cgagtcagtg agcgagggag cggaagagcg cccaatacgc
     5641 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     5701 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     5761 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     5821 caatttcaca caggaaacag ctatgaccat gattacgcca agctgctcga gaatgaaaga
     5881 cccccgaggt gggcagtcaa tcactcagag gagaccctcc caaggaacag cgagaccacg
     5941 agtcggatgc aactgcaaga gggtttattg agaacacggg tacccgggcg acgcagtcta
     6001 tcggaggact ggcgcgccg
//
