LOCUS       sequence-357081-        8487 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mouse full length TGF-beta-3 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8487)
  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 8487)
  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-Flag-mouse TGFb3
FEATURES             Location/Qualifiers
     source          1..8487
                     /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..1808
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    4558..4613
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4612..4631)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(5323..5911)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5403..5422)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5997..6806)
                     /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     6720..6739
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(6825..6872)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             complement(7298..7897)
                     /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(7381..7401)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     7878..7897
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(7907..8406)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7955..7974)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     8348..8371
                     /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 ggccgactac aaggacgacg
     1801 acgacaagct cgagacttgc accacgttgg acttcggcca catcaagaag aagagggtgg
     1861 aagccattag gggacagatc ttgagcaagc tcaggctcac cagcccccct gagccatcgg
     1921 tgatgaccca cgtcccctat caggtcctgg cactttacaa cagcacccgg gagttgctgg
     1981 aagagatgca cggggagagg gaggaaggct gcactcagga gacctcggag tctgagtact
     2041 atgccaaaga gatccataaa ttcgacatga tccagggact ggcggagcac aatgaactgg
     2101 ccgtctgccc caaaggaatt acctctaagg tttttcgttt caatgtgtcc tcagtggaga
     2161 aaaatggaac caatctgttc cgggcagagt tccgggtctt gcgggtgccc aaccccagct
     2221 ccaagcgcac agagcagaga attgagctct tccagatact tcgaccggat gagcacatag
     2281 ccaagcagcg ctacataggt ggcaagaatc tgcccacaag gggcaccgct gaatggctgt
     2341 ctttcgatgt cactgacact gtgcgcgagt ggctgttgag gagagagtcc aacttgggtc
     2401 tggaaatcag catccactgt ccatgtcaca cctttcagcc caatggagac atactggaaa
     2461 atgttcatga ggtgatggaa atcaaattca aaggagtgga caatgaagat gaccatggcc
     2521 gtggagacct ggggcgtctc aagaagcaaa aggatcacca caacccacac ctgatcctca
     2581 tgatgatccc cccacaccga ctggacagcc caggccaggg cagtcagagg aagaagaggg
     2641 ccctggacac caattactgc ttccgcaacc tggaggagaa ctgctgtgta cgcccccttt
     2701 atattgactt ccggcaggat ctaggctgga aatgggtcca cgaacctaag ggttactatg
     2761 ccaacttctg ctcaggccct tgcccatacc tccgcagcgc agacacaacc catagcacgg
     2821 tgcttggact atacaacacc ctgaacccag aggcgtctgc ctcgccatgc tgcgtccccc
     2881 aggacctgga gcccctgacc atcttgtact atgtgggcag aacccccaag gtggagcagc
     2941 tgtccaacat ggtggtgaag tcgtgtaagt gcagctgata agctagcggt tgaggtctct
     3001 aaaagcgtct tcctgttctc atcacatcat atcaaggtta tataccatca atattgccac
     3061 agatgttact tagcctttta atatttctct aatttagtgt atatgcaatg atagttctct
     3121 gatttctgag attgagtttc tcatgtgtaa tgattattta gagtttctct ttcatctgtt
     3181 caaatttttg tctagtttta ttttttactg atttgtaaga cttcttttta taatctgcat
     3241 attacaattc tctttactgg ggtgttgcaa atattttctg tcattctatg gcctgacttt
     3301 tcttaatggt tttttaattt taaaaataag tcttaatatt catgcaatct aattaacaat
     3361 cttttctttg tggttaggac tttgagtcat aagaaatttt tctctacact gaagtcatga
     3421 tggcatgctt ctatattatt ttctaaaaga tttaaagttt tgccttctcc atttagactt
     3481 ataattcact ggaatttttt tgtgtgtatg gtatgacata tgggttccct tttatttttt
     3541 acatataaat atatttccct gtttttctaa aaaagaaaaa gatcatcatt ttcccattgt
     3601 aaaatgccat atttttttca taggtcactt acatatatca atgggtctgt ttctgagctc
     3661 tactctattt tatcagcctc actgtctatc cccacacatc tcatgctttg ctctaaatct
     3721 tgatatttag tggaacattc tttcccattt tgttctacaa gaatattttt gttattgtct
     3781 ttgggctttc tatatacatt ttgaaatgag gttgacaagt taataacagg cctattgatt
     3841 ggaaagtttg tcaacgaatt gtgggtcttt tggggtttgc tgcccctttt acgcaatgtg
     3901 gatatcctgc tttaatgcct ttatatgcat gtatacaagc aaaacaggct tttactttct
     3961 cgccaactta caaggccttt ctcagtaaac agtatatgac cctttacccc gttgctcggc
     4021 aacggcctgg tctgtgccaa gtgtttgctg acgcaacccc cactggttgg ggcttggcca
     4081 taggccatca gcgcatgcgt ggaacctttg tgtctcctct gccgatccat actgcggaac
     4141 tcctagccgc ttgttttgct cgcagcaggt ctggagcaaa cctcatcggg accgacaatt
     4201 ctgtcgtact ctcccgcaag tatacatcgt ttccatggct gctaggctgt gctgccaact
     4261 ggatcctgcg cgggacgtcc tttgtttacg tcccgtcggc gctgaatccc gcggacgacc
     4321 cctcccgggg ccgcttgggg ctctaccgcc cgcttctccg tctgccgtac cgtccgacca
     4381 cggggcgcac ctctctttac gcggactccc cgtctgtgcc ttctcatctg ccggaccgtg
     4441 tgcacttcgc ttcacctctg cacgtcgcat ggaggccacc gtgaacgccc accggaacct
     4501 gcccaaggtc ttgcataaga ggactcttgg actttcagca atgtcatctc agtggctaat
     4561 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     4621 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     4681 caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt
     4741 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt
     4801 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct
     4861 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc
     4921 ctcttctctt atggagatct ttgcattttt agacatttag aagcctatat cttgttacag
     4981 aattggaatt acacaaaaat tctaccatat tttgaaagct taggttgttc tgaaaaaaac
     5041 aatatattgt tttcctgggt aaactaaaag tcccctcgag gaaaggcccc taaagtgaaa
     5101 cagtgcaaaa cgttcaaaaa ctgtctggca atacaagttc cactttgacc aaaacggctg
     5161 gcagtaaaag ggttaagaag actgtcagcc ttgagcggta tcagctcact caaaggcggt
     5221 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5281 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5341 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5401 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5461 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5521 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5581 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5641 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5701 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg gctaactacg gctacactag
     5761 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5821 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5881 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5941 tgacgctcag tggaacgacg cgcgcgtaac tcacgttaag ggattttggt catgagttag
     6001 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca
     6061 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg
     6121 atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt
     6181 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa
     6241 tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca
     6301 ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc
     6361 tgagcgaggc gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgagtgc
     6421 aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct
     6481 tctaatacct ggaacgctgt ttttccgggg atcgcagtgg tgagtaacca tgcatcatca
     6541 ggagtacgga taaaatgctt gatggtcgga agtggcataa attccgtcag ccagtttagt
     6601 ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac
     6661 tctggcgcat cgggcttccc atacaagcga tagattgtcg cacctgattg cccgacatta
     6721 tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcctc
     6781 gacgtttccc gttggatatg gctcattttt tacttcctca ccttgtcgta ttatactatg
     6841 ccgatatact atgccgatga ttaattgtcg acactgcggg ggctctgtgt ggtaagcagg
     6901 tcttaacctt tttactgcca atgacgcatg ggatacgtcg tggcagtaaa agggcttaaa
     6961 tgccaacgac gcgtcccata cgttgttggc attttaattc ttctctctgc agcggcagca
     7021 tgtgccgccg ctgcagagag tttctagcga tgacagcccc tctgggcaac gagccggggg
     7081 ggctgtcttt ctttatgttt taaatgcact gacctcccac attccctttt tagtaaaata
     7141 ttcagaaata atttaaatac atcattgcaa tgaaaataaa tgttttttat taggcagaat
     7201 ccagatgctc aaggcccttc ataatatccc ccagtttagt agttggactt agggaacaaa
     7261 ggaaccttta atagaaattg gacagcaaga aagcgagtca ggcaccgggc ttgcgggtca
     7321 tgcaccaggt gcgcggtcct tcgggcacct cgacgtcggc ggtgacggtg aagccgagcc
     7381 gctcgtagaa ggggaggttg cggggcgcgg atgtctccag gaaggcgggc accccggcgc
     7441 gctcggccgc ctccactccg gggagcacga cggcgctgcc cagacccttg ccctggtggt
     7501 cgggcgacac gccgacggtg gccaggaacc acgcgggctc cttgggccgg tgcggcgcca
     7561 ggaggccttc catctgttgc tgcgcggcca gccgggaacc gctcaactcg gccatgcgcg
     7621 ggccgatctc ggcgaacacc gcccccgctt cgacgctctc cggcgtggtc cagaccgcca
     7681 ccgcggcgcc gtcgtccgcg acccacacct tgccgatgtc gagcccgacg cgcgtgagga
     7741 agagttcttg cagctcggtg acccgctcga tgtggcggtc cggatcgacg gtgtggcgcg
     7801 tggcggggta gtcggcgaac gcggcggcga gggtgcgtac ggccctgggg acgtcgtcgc
     7861 gggtggcgag gcgcaccgtg ggcttgtact cggtcatggt ggcggacgaa aggcccggag
     7921 atgaggaaga ggagaacagc gcggcagacg tgcgcttttg aagcgtgcag aatgccgggc
     7981 ctccggagga ccttcgggcg cccgccccgc ccctgagccc gcccctgagc ccgcccccgg
     8041 acccacccct tcccagcctc tgagcccaga aagcgaagga gcaaagctgc tattggccgc
     8101 tgccccaaag gcctacccgc ttccattgct cagcggtgct gtccatctgc acgagactag
     8161 tgagtcgtgc tacttccatt tgtcacgtcc tgcacgacgc gagctgcggg gcggggggga
     8221 acttcctgac taggggagga gtagaaggtg gcgcgaaggg gccaccaaag aacggagccg
     8281 gttggcgcct accggtggat gtggaatgtg tgcgaggcca gaggccactt gtgtagcgcc
     8341 aagtgcccag cggggctgct aaagcgcatg ctccagactg ccttgggaaa agcgcctccc
     8401 ctacccggta gagaaacttg atctgtcgcc gcaattcaaa cttcgtgagg ctccggtgcc
     8461 cgtcagtgac ctgctatact ctggaga
//
