LOCUS       sequence-357201-        9267 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mouse full length GARP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9267)
  AUTHORS   Jiang A, Qin Y, Springer TA
  TITLE     Loss of LRRC33-dependent TGFbeta1 activation enhances anti-tumor 
            immunity and checkpoint blockade therapy.
  JOURNAL   Cancer Immunol Res. 2022 Feb 18. pii: 681626. doi: 
            10.1158/2326-6066.CIR-21-0593.
  PUBMED    35181792
REFERENCE   2  (bases 1 to 9267)
  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: "Cancer 
            Immunol Res. 2022 Feb 18. pii: 681626. doi: 
            10.1158/2326-6066.CIR-21-0593."
COMMENT     Sequence Label: pD2529-CAG-HA-mouse GARP
FEATURES             Location/Qualifiers
     source          1..9267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        3..306
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        308..583
                     /label=chicken beta-actin promoter
     intron          584..1600
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1524..1546
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1608..1627
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1785..1811
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    5338..5393
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5392..5411)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(6103..6691)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6183..6202)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6777..7586)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     7500..7519
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(7605..7652)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             complement(8078..8677)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(8161..8181)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     8658..8677
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(8687..9186)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(8735..8754)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     9128..9151
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
       61 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      121 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      181 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      241 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      301 accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca
      361 cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg
      421 gggggggcgc gcgccaggcg gggcggggsg gggsgrgggg sggggsgggg sgrggcggrg
      481 aggtgcggcg gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg
      541 gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg
      601 ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac
      661 cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg
      721 cttggtttaa tgacggcttg tttcttttct gtggctgcgt gaaagccttg aggggctccg
      781 ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg
      841 gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg
      901 gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg
      961 cggggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc
     1021 agggggtgtg ggcgcgtcgg tcgggctgca accccccctg cacccccctc cccgagttgc
     1081 tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt
     1141 gccgggcggg gggtggcggc aggtgggggt gccgggcggg gcggggccgc ctcgggccgg
     1201 ggagggctcg ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc
     1261 gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc
     1321 ccaaatctgt gcggagccga aatctgggag gcgccgccgc accccctcta gcgggcgcgg
     1381 ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc
     1441 gcgccgccgt ccccttctcc ctctccagcc tcggggctgt ccgcgggggg acggctgcct
     1501 tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg gctctagagc
     1561 ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt
     1621 tattgtgctg tctcatcatt ttggcaaaga attcggcttg atcgaagccg tctcagggga
     1681 gagaccacac ccaagctgtc tagagccgcc accttgtgaa gagcgccgcc accatgcctt
     1741 tgctcctcct gttgcccttg ctctgggctg gcgccctcgc ggcctaccca tacgatgttc
     1801 cagattacgc tctcgagatc tctcagcgtc gagagcaagt gccctgtagg acggtgaaca
     1861 aggaggcctt gtgccacggc cttggcctgc tccaggtccc ctcggtgctc tcactggaca
     1921 tccaagctct ctacttgtcc gggaaccaac tgcagagcat cctggtctcc ccactgggct
     1981 tctatacagc gcttcgtcac ctggatttaa gtgacaacca gattagcttc ctccaggctg
     2041 gggtcttcca ggccctgccc tacctggagc accttaacct ggcccacaat cggcttgcca
     2101 caggcatggc tctcaacagt ggtggtctgg gccgcctgcc ccttttggtc tccctggacc
     2161 tgtctgggaa cagcctgcat ggcaatctgg tggagcgact gttgggtgag accccgaggc
     2221 tgcgtacgct ctcactggcg gagaacagcc tcactcgcct ggcacgccac accttctggg
     2281 gtatgccagc ggtggagcaa ttagacctcc acagcaatgt cctcatggac atcgaggacg
     2341 gtgcctttga ggccctgccc cacctgactc acctcaatct ctccagaaac tccctcacct
     2401 gcatctcaga cttcagcctc cagcagctgc aggtactaga cttgagctgc aacagcattg
     2461 aggccttcca gacggccccg gaaccacagg cccagttcca gttggcctgg ctcgacctcc
     2521 gggagaacaa gctgctccac ttccctgacc tggccgtgtt cccgagactc atctacctga
     2581 atgtgtctaa caacctcatc cagctccctg cggggctgcc ccggggcagt gaggacctcc
     2641 acgcaccctc tgaaggctgg tcagcctctc cactgtccaa ccccagccgg aatgccagca
     2701 cccaccctct ctcccagctc ctgaacctgg atctgagtta caatgagatc gaactggtcc
     2761 ctgctagctt tcttgaacac ctgacctccc tgcgcttcct caacctcagc agaaactgcc
     2821 tgcgatcctt tgaggctcga caagtggact ccctgccctg cctggtgctt ttggacttga
     2881 gccacaatgt gctggaagca ttagaactgg gtaccaaagt cctagggtcc ctgcagacgt
     2941 tgcttctgca ggacaatgcc ttgcaagaac ttccacccta tacctttgcc agcctggcca
     3001 gcctgcagag gctaaaccta caggggaacc aggtcagtcc ctgtgggggg ccagcagaac
     3061 caggtccccc gggctgtgtg gacttctctg ggatccccac ccttcatgtt cttaacatgg
     3121 cagggaactc gatgggaatg ctcagggcgg gcagcttcct tcacacccca cttacagagc
     3181 tggacctctc caccaatcct ggtctggatg tggccacggg agccctggtg ggcctggagg
     3241 catccttaga ggtactggag cttcaaggca acgggctgac tgtcttgaga gtggacttgc
     3301 cctgctttct ccgtctcaag cgccttaacc ttgctgagaa ccagttaagc cacctgcccg
     3361 cttggacgcg ggctgtgtcc ctggaggtgc tagatctgcg gaacaacagc ttcagcctcc
     3421 tgccaggcaa tgccatgggt ggcctggaga ccagtctccg gcggctgtac ttgcagggaa
     3481 atccactcag ctgctgtggc aatggctggc tggcggccca gctgcaccag ggcagagtgg
     3541 atgtggatgc tactcaggac ctaatctgcc gctttggctc ccaggaggag ttgtccctga
     3601 gcctagtgcg tccagaggat tgcgagaagg gagggctgaa gaatgtcaac ctcatcctcc
     3661 tcctcagctt cacactggtc tctgccatcg tcctcaccac gctggccacc atctgcttcc
     3721 tccgccggca gaagctcagc caacaataca aagcctgata agctagcggt tgaggtctct
     3781 aaaagcgtct tcctgttctc atcacatcat atcaaggtta tataccatca atattgccac
     3841 agatgttact tagcctttta atatttctct aatttagtgt atatgcaatg atagttctct
     3901 gatttctgag attgagtttc tcatgtgtaa tgattattta gagtttctct ttcatctgtt
     3961 caaatttttg tctagtttta ttttttactg atttgtaaga cttcttttta taatctgcat
     4021 attacaattc tctttactgg ggtgttgcaa atattttctg tcattctatg gcctgacttt
     4081 tcttaatggt tttttaattt taaaaataag tcttaatatt catgcaatct aattaacaat
     4141 cttttctttg tggttaggac tttgagtcat aagaaatttt tctctacact gaagtcatga
     4201 tggcatgctt ctatattatt ttctaaaaga tttaaagttt tgccttctcc atttagactt
     4261 ataattcact ggaatttttt tgtgtgtatg gtatgacata tgggttccct tttatttttt
     4321 acatataaat atatttccct gtttttctaa aaaagaaaaa gatcatcatt ttcccattgt
     4381 aaaatgccat atttttttca taggtcactt acatatatca atgggtctgt ttctgagctc
     4441 tactctattt tatcagcctc actgtctatc cccacacatc tcatgctttg ctctaaatct
     4501 tgatatttag tggaacattc tttcccattt tgttctacaa gaatattttt gttattgtct
     4561 ttgggctttc tatatacatt ttgaaatgag gttgacaagt taataacagg cctattgatt
     4621 ggaaagtttg tcaacgaatt gtgggtcttt tggggtttgc tgcccctttt acgcaatgtg
     4681 gatatcctgc tttaatgcct ttatatgcat gtatacaagc aaaacaggct tttactttct
     4741 cgccaactta caaggccttt ctcagtaaac agtatatgac cctttacccc gttgctcggc
     4801 aacggcctgg tctgtgccaa gtgtttgctg acgcaacccc cactggttgg ggcttggcca
     4861 taggccatca gcgcatgcgt ggaacctttg tgtctcctct gccgatccat actgcggaac
     4921 tcctagccgc ttgttttgct cgcagcaggt ctggagcaaa cctcatcggg accgacaatt
     4981 ctgtcgtact ctcccgcaag tatacatcgt ttccatggct gctaggctgt gctgccaact
     5041 ggatcctgcg cgggacgtcc tttgtttacg tcccgtcggc gctgaatccc gcggacgacc
     5101 cctcccgggg ccgcttgggg ctctaccgcc cgcttctccg tctgccgtac cgtccgacca
     5161 cggggcgcac ctctctttac gcggactccc cgtctgtgcc ttctcatctg ccggaccgtg
     5221 tgcacttcgc ttcacctctg cacgtcgcat ggaggccacc gtgaacgccc accggaacct
     5281 gcccaaggtc ttgcataaga ggactcttgg actttcagca atgtcatctc agtggctaat
     5341 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     5401 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     5461 caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt
     5521 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt
     5581 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct
     5641 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc
     5701 ctcttctctt atggagatct ttgcattttt agacatttag aagcctatat cttgttacag
     5761 aattggaatt acacaaaaat tctaccatat tttgaaagct taggttgttc tgaaaaaaac
     5821 aatatattgt tttcctgggt aaactaaaag tcccctcgag gaaaggcccc taaagtgaaa
     5881 cagtgcaaaa cgttcaaaaa ctgtctggca atacaagttc cactttgacc aaaacggctg
     5941 gcagtaaaag ggttaagaag actgtcagcc ttgagcggta tcagctcact caaaggcggt
     6001 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6061 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6121 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6181 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6241 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6301 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6361 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6421 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6481 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg gctaactacg gctacactag
     6541 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6601 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6661 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6721 tgacgctcag tggaacgacg cgcgcgtaac tcacgttaag ggattttggt catgagttag
     6781 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca
     6841 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg
     6901 atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt
     6961 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa
     7021 tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca
     7081 ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc
     7141 tgagcgaggc gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgagtgc
     7201 aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct
     7261 tctaatacct ggaacgctgt ttttccgggg atcgcagtgg tgagtaacca tgcatcatca
     7321 ggagtacgga taaaatgctt gatggtcgga agtggcataa attccgtcag ccagtttagt
     7381 ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac
     7441 tctggcgcat cgggcttccc atacaagcga tagattgtcg cacctgattg cccgacatta
     7501 tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcctc
     7561 gacgtttccc gttggatatg gctcattttt tacttcctca ccttgtcgta ttatactatg
     7621 ccgatatact atgccgatga ttaattgtcg acactgcggg ggctctgtgt ggtaagcagg
     7681 tcttaacctt tttactgcca atgacgcatg ggatacgtcg tggcagtaaa agggcttaaa
     7741 tgccaacgac gcgtcccata cgttgttggc attttaattc ttctctctgc agcggcagca
     7801 tgtgccgccg ctgcagagag tttctagcga tgacagcccc tctgggcaac gagccggggg
     7861 ggctgtcttt ctttatgttt taaatgcact gacctcccac attccctttt tagtaaaata
     7921 ttcagaaata atttaaatac atcattgcaa tgaaaataaa tgttttttat taggcagaat
     7981 ccagatgctc aaggcccttc ataatatccc ccagtttagt agttggactt agggaacaaa
     8041 ggaaccttta atagaaattg gacagcaaga aagcgagtca ggcaccgggc ttgcgggtca
     8101 tgcaccaggt gcgcggtcct tcgggcacct cgacgtcggc ggtgacggtg aagccgagcc
     8161 gctcgtagaa ggggaggttg cggggcgcgg atgtctccag gaaggcgggc accccggcgc
     8221 gctcggccgc ctccactccg gggagcacga cggcgctgcc cagacccttg ccctggtggt
     8281 cgggcgacac gccgacggtg gccaggaacc acgcgggctc cttgggccgg tgcggcgcca
     8341 ggaggccttc catctgttgc tgcgcggcca gccgggaacc gctcaactcg gccatgcgcg
     8401 ggccgatctc ggcgaacacc gcccccgctt cgacgctctc cggcgtggtc cagaccgcca
     8461 ccgcggcgcc gtcgtccgcg acccacacct tgccgatgtc gagcccgacg cgcgtgagga
     8521 agagttcttg cagctcggtg acccgctcga tgtggcggtc cggatcgacg gtgtggcgcg
     8581 tggcggggta gtcggcgaac gcggcggcga gggtgcgtac ggccctgggg acgtcgtcgc
     8641 gggtggcgag gcgcaccgtg ggcttgtact cggtcatggt ggcggacgaa aggcccggag
     8701 atgaggaaga ggagaacagc gcggcagacg tgcgcttttg aagcgtgcag aatgccgggc
     8761 ctccggagga ccttcgggcg cccgccccgc ccctgagccc gcccctgagc ccgcccccgg
     8821 acccacccct tcccagcctc tgagcccaga aagcgaagga gcaaagctgc tattggccgc
     8881 tgccccaaag gcctacccgc ttccattgct cagcggtgct gtccatctgc acgagactag
     8941 tgagtcgtgc tacttccatt tgtcacgtcc tgcacgacgc gagctgcggg gcggggggga
     9001 acttcctgac taggggagga gtagaaggtg gcgcgaaggg gccaccaaag aacggagccg
     9061 gttggcgcct accggtggat gtggaatgtg tgcgaggcca gaggccactt gtgtagcgcc
     9121 aagtgcccag cggggctgct aaagcgcatg ctccagactg ccttgggaaa agcgcctccc
     9181 ctacccggta gagaaacttg atctgtcgcc gcaattcaaa cttcgtgagg ctccggtgcc
     9241 cgtcagtgac ctgctatact ctggaga
//
