LOCUS       sequence-349385-        5908 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses secreted murine GM-CSF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5908)
  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 5908)
  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-GM-CSF
FEATURES             Location/Qualifiers
     source          1..5908
                     /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    2363..2766
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             3272..3862
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3887..3903)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3887..3903)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3900..3922)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3911..3927
                     /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(3935..3941,3942..3959,3960..3965))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3980..4001
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4118..4135)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4289..4877)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4369..4388)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5048..5839,5840..5908))
                     /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     5671..5690
                     /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 ctgaccagat tcctgagcct
     1741 gaacctgctg ctgctgggcg agtctatcat cctcggatct ggcgaggccg ctcctaccag
     1801 atctcctatc accgtgacca gaccttggaa gcacgtggaa gccatcaaag aggccctgaa
     1861 tctgctggac gacatgcccg tgacactgaa cgaagaggtg gaagtggtgt ccaacgagtt
     1921 cagcttcaag aaactgacct gcgtgcagac ccggctgaag atctttgagc agggcctgag
     1981 aggcaacttc accaagctga agggcgctct gaacatgacc gccagctact accagaccta
     2041 ctgtcctcct acacctgaga cagactgcga gacacaagtg accacctacg ccgacttcat
     2101 cgacagcctg aaaaccttcc tgacagacat ccccttcgag tgcaagaaac ccggccagaa
     2161 gtgagaattc gagcatctta ccgccattta ttcccatatt tgttctgttt ttcttgattt
     2221 gggtatacat ttaaatgtta ataaaacaaa atggtggggc aatcatttac attttatggg
     2281 atatgtaatt actagttcag gtgtattgcc acaagacaaa catgttaaga aactttcccg
     2341 ttatttacgc tctgttcctg ttaatcaacc tctggattac aaaatttgtg aaagattgac
     2401 tgatattctt aactatgttg ctccttttac gctgtgtgga tatgctgctt taatgcctct
     2461 gtatcatgct attgcttccc gtacggcttt cgttttctcc tccttgtata aatcctggtt
     2521 gctgtctctt tatgaggagt tgtggcccgt tgtccgtcaa cgtggcgtgg tgtgctctgt
     2581 gtttgctgac gcaaccccca ctggctgggg cattgccacc acctgtcaac tcctttctgg
     2641 gactttcgct ttccccctcc cgatcgccac ggcagaactc atcgccgcct gccttgcccg
     2701 ctgctggaca ggggctaggt tgctgggcac tgataattcc gtggtgttgt cggggaagct
     2761 gacgtccttt ccatggctgc tcgcctgtgt tgccaactgg atcctgcgcg ggacgtcctt
     2821 ctgctacgtc ccttcggctc tcaatccagc ggacctccct tcccgaggcc ttctgccggt
     2881 tctgcggcct ctcccgcgtc ttcgctttcg gcctccgacg agtcggatct ccctttgggc
     2941 cgcctccccg cctgtttcgc ctcggcgtcc ggtccgtgtt gcttggtcgt cacctgtgca
     3001 gaattgcgaa ccatggattc caccgtgaac tttgtctcct ggcatgcaaa tcgtcaactt
     3061 ggcatgccaa gaataattcg gatccaagct taggcctgct cgctttcttg ctgtcccatt
     3121 tctattaaag gttcctttgt tccctaagtc caactactaa actgggggat attatgaagg
     3181 gccttgagca tctggattct gcctagcgct aagcttaaca cgagccatag atagaataaa
     3241 agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca
     3301 agctagctta agtaacgcca ttttgcaagg catggaaaat acataactga gaatagagaa
     3361 gttcagatca aggttaggaa cagagagaca gcagaatatg ggccaaacag gatatctgtg
     3421 gtaagcagtt cctgccccgg ctcagggcca agaacagttg gaacagcaga atatgggcca
     3481 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc
     3541 cagatgcggt cccgccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc
     3601 aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc
     3661 tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggcgc
     3721 gccagtcctc cgatagactg cgtcgcccgg gtacccgtgt tctcaataaa ccctcttgca
     3781 gttgcatccg actcgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgactg
     3841 cccacctcgg gggtctttca ttctcgagca gcttggcgta atcatggtca tagctgtttc
     3901 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     3961 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4021 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4081 ggagaggcgg tttgcgtatt gggcgctctt ccgctccctc gctcactgac tcgctgcgct
     4141 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4201 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4261 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4321 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4381 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4441 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4501 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4561 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4621 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4681 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4741 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4801 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4861 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4921 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4981 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5041 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5101 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5161 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5221 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5281 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5341 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5401 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5461 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5521 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5581 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5641 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5701 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5761 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5821 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5881 gggcgacacg gaaatgttga atactcat
//
