LOCUS       sequence-344461-        5935 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of secreted murine IFN gamma.
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-IFN-gamma
FEATURES             Location/Qualifiers
     source          1..5935
                     /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    1719..2122
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             2628..3218
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3243..3259)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3243..3259)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3256..3278)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3267..3283
                     /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(3291..3297,3298..3315,3316..3321))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3336..3357
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3474..3491)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3645..4233)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3725..3744)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4404..5195,5196..5264))
                     /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     5027..5046
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5265..5369)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5437..5455
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5493..5515)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5615..5634
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5828..5850
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5842..5859
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5843..5859
                     /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 gctagccctc tgacaagatt cctgagcctg aacctgctgc
     1081 tgctgggcga gtctatcatc ctcggatctg gcgaggctca cggcaccgtg atcgagtctc
     1141 tggaaagcct gaacaactac ttcaacagca gcggcatcga cgtggaagag aagtccctgt
     1201 tcctggacat ctggcggaac tggcagaaag acggcgacat gaagatcctg cagagccaga
     1261 tcatcagctt ctacctgaga ctgttcgagg tgctgaagga caaccaggcc atcagcaaca
     1321 acatctctgt gatcgagagc cacctgatca ccacattctt cagcaacagc aaggccaaga
     1381 aagacgcctt catgtctatc gccaagttcg aagtgaacaa ccctcaggtg cagagacagg
     1441 ccttcaacga gctgatcaga gtggtgcatc agctgctgcc agagagcagc ctgagaaagc
     1501 gcaagagaag cagatgctag gaattcgagc atcttaccgc catttattcc catatttgtt
     1561 ctgtttttct tgatttgggt atacatttaa atgttaataa aacaaaatgg tggggcaatc
     1621 atttacattt tatgggatat gtaattacta gttcaggtgt attgccacaa gacaaacatg
     1681 ttaagaaact ttcccgttat ttacgctctg ttcctgttaa tcaacctctg gattacaaaa
     1741 tttgtgaaag attgactgat attcttaact atgttgctcc ttttacgctg tgtggatatg
     1801 ctgctttaat gcctctgtat catgctattg cttcccgtac ggctttcgtt ttctcctcct
     1861 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc cgtcaacgtg
     1921 gcgtggtgtg ctctgtgttt gctgacgcaa cccccactgg ctggggcatt gccaccacct
     1981 gtcaactcct ttctgggact ttcgctttcc ccctcccgat cgccacggca gaactcatcg
     2041 ccgcctgcct tgcccgctgc tggacagggg ctaggttgct gggcactgat aattccgtgg
     2101 tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc aactggatcc
     2161 tgcgcgggac gtccttctgc tacgtccctt cggctctcaa tccagcggac ctcccttccc
     2221 gaggccttct gccggttctg cggcctctcc cgcgtcttcg ctttcggcct ccgacgagtc
     2281 ggatctccct ttgggccgcc tccccgcctg tttcgcctcg gcgtccggtc cgtgttgctt
     2341 ggtcgtcacc tgtgcagaat tgcgaaccat ggattccacc gtgaactttg tctcctggca
     2401 tgcaaatcgt caacttggca tgccaagaat aattcggatc caagcttagg cctgctcgct
     2461 ttcttgctgt cccatttcta ttaaaggttc ctttgttccc taagtccaac tactaaactg
     2521 ggggatatta tgaagggcct tgagcatctg gattctgcct agcgctaagc ttaacacgag
     2581 ccatagatag aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc
     2641 acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaatacat
     2701 aactgagaat agagaagttc agatcaaggt taggaacaga gagacagcag aatatgggcc
     2761 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagttggaac
     2821 agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     2881 aagaacagat ggtccccaga tgcggtcccg ccctcagcag tttctagaga accatcagat
     2941 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     3001 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
     3061 acccctcact cggcgcgcca gtcctccgat agactgcgtc gcccgggtac ccgtgttctc
     3121 aataaaccct cttgcagttg catccgactc gtggtctcgc tgttccttgg gagggtctcc
     3181 tctgagtgat tgactgccca cctcgggggt ctttcattct cgagcagctt ggcgtaatca
     3241 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     3301 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     3361 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     3421 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc tccctcgctc
     3481 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3541 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3601 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3661 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3721 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3781 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3841 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3901 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3961 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4021 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4081 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4141 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4201 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4261 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4321 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4381 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4441 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     4501 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     4561 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4621 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4681 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4741 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     4801 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     4861 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4921 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4981 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     5041 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     5101 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     5161 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     5221 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     5281 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     5341 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     5401 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     5461 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     5521 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     5581 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     5641 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     5701 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     5761 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     5821 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattagtact ctagcttaag
     5881 taacgccatt ttgcaaggca tggaaaatac ataactgaga atagagaagt tcaga
//
