LOCUS       sequence-343876-        6025 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Secretes the extracellular domain of murine Flt3L.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6025)
  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 6025)
  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-Flt3L
FEATURES             Location/Qualifiers
     source          1..6025
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(41..145)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     213..231
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(269..291)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     391..410
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     604..626
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     618..635
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     619..635
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      1743..1752
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2520..2923
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             3429..4019
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4044..4060)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4044..4060)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4057..4079)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4068..4084
                     /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(4092..4098,4099..4116,4117..4122))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4137..4158
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4275..4292)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4446..5034)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4526..4545)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5205..5996,5997..6025,1..40))
                     /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     5828..5847
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
       61 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
      121 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
      181 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
      241 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
      301 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      361 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      421 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      481 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      541 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      601 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt agtactctag cttaagtaac
      661 gccattttgc aaggcatgga aaatacataa ctgagaatag agaagttcag atcaaggtta
      721 ggaacagaga gacagcagaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
      781 ccggctcagg gccaagaaca gttggaacag cagaatatgg gccaaacagg atatctgtgg
      841 taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtcccgcc
      901 ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac
      961 cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg
     1021 ctccccgagc tcaataaaag agcccacaac ccctcactcg gcgcgccagt cctccgatag
     1081 actgcgtcgc ccgggtaccc gtattcccaa taaagcctct tgctgtttgc atccgaatcg
     1141 tggactcgct gatccttggg agggtctcct cagattgatt gactgcccac ctcgggggtc
     1201 tttcatttgg aggttccacc gagatttgga gacccctgcc cagggaccac cgaccccccc
     1261 gccgggaggt aagctggcca gcggtcgttt cgtgtctgtc tctgtctttg tgcgtgtttg
     1321 tgccggcatc taatgtttgc gcctgcgtct gtactagttg gctaactaga tctgtatctg
     1381 gcggtcccgc ggaagaactg acgagttcgt attcccggcc gcagcccctg ggagacgtcc
     1441 cagcggcctc gggggcccgt tttgtggccc attctgtatc agttaaccta cccgagtcgg
     1501 actttttgga gctccgccac tgtccgaggg gtacgtggct ttgttggggg acgagagaca
     1561 gagacacttc ccgcccccgt ctgaattttt gctttcggtt ttacgccgaa accgcgccgc
     1621 gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt
     1681 gtctgaaaat tagctcgaca aagttaagta atagtccctc tctccaagct cacttacagg
     1741 cggccgccat ggctagccct ctgacaagat tcctgagcct gaacctgctg ctgctgggcg
     1801 agtctatcat cctcggatct ggcgaggctg gcacccctga ctgttacttc agccactctc
     1861 ctatcagcag caacttcaaa gtgaagttcc gcgagctgac cgaccatctg ctgaaggact
     1921 accctgtgac cgtggccgtg aacctgcagg atgagaagca ctgcaaggcc ctgtggtccc
     1981 tgttcctggc tcagagatgg atcgagcagc tgaaaacagt ggccggcagc aagatgcaga
     2041 ccctgctgga agatgtgaac accgagatcc acttcgtgac cagctgcacc ttccagcctc
     2101 tgcctgagtg cctgagattc gtgcagacca acatcagcca ccttctcaag gacacatgca
     2161 cccagctgct ggccctgaag ccttgtatcg gcaaggcctg ccagaacttc tccagatgcc
     2221 tggaagtgca gtgccagcca gacagctcta cactgctgcc tccaagaagc cctatcgctc
     2281 tggaagccac agagctgcct gagcctagac ctagacagta ggaattcgag catcttaccg
     2341 ccatttattc ccatatttgt tctgtttttc ttgatttggg tatacattta aatgttaata
     2401 aaacaaaatg gtggggcaat catttacatt ttatgggata tgtaattact agttcaggtg
     2461 tattgccaca agacaaacat gttaagaaac tttcccgtta tttacgctct gttcctgtta
     2521 atcaacctct ggattacaaa atttgtgaaa gattgactga tattcttaac tatgttgctc
     2581 cttttacgct gtgtggatat gctgctttaa tgcctctgta tcatgctatt gcttcccgta
     2641 cggctttcgt tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     2701 ggcccgttgt ccgtcaacgt ggcgtggtgt gctctgtgtt tgctgacgca acccccactg
     2761 gctggggcat tgccaccacc tgtcaactcc tttctgggac tttcgctttc cccctcccga
     2821 tcgccacggc agaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctaggttgc
     2881 tgggcactga taattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg
     2941 cctgtgttgc caactggatc ctgcgcggga cgtccttctg ctacgtccct tcggctctca
     3001 atccagcgga cctcccttcc cgaggccttc tgccggttct gcggcctctc ccgcgtcttc
     3061 gctttcggcc tccgacgagt cggatctccc tttgggccgc ctccccgcct gtttcgcctc
     3121 ggcgtccggt ccgtgttgct tggtcgtcac ctgtgcagaa ttgcgaacca tggattccac
     3181 cgtgaacttt gtctcctggc atgcaaatcg tcaacttggc atgccaagaa taattcggat
     3241 ccaagcttag gcctgctcgc tttcttgctg tcccatttct attaaaggtt cctttgttcc
     3301 ctaagtccaa ctactaaact gggggatatt atgaagggcc ttgagcatct ggattctgcc
     3361 tagcgctaag cttaacacga gccatagata gaataaaaga ttttatttag tctccagaaa
     3421 aaggggggaa tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt
     3481 tgcaaggcat ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag
     3541 agagacagca gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc
     3601 agggccaaga acagttggaa cagcagaata tgggccaaac aggatatctg tggtaagcag
     3661 ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtccc gccctcagca
     3721 gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
     3781 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
     3841 agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga tagactgcgt
     3901 cgcccgggta cccgtgttct caataaaccc tcttgcagtt gcatccgact cgtggtctcg
     3961 ctgttccttg ggagggtctc ctctgagtga ttgactgccc acctcggggg tctttcattc
     4021 tcgagcagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     4081 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     4141 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     4201 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4261 cgctcttccg ctccctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4321 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4381 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4441 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4501 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4561 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4621 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4681 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4741 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4801 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4861 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4921 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4981 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5041 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5101 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5161 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5221 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5281 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5341 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5401 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5461 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5521 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5581 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5641 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5701 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5761 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5821 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5881 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5941 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6001 aaaacaggaa ggcaaaatgc cgcaa
//
