LOCUS       sequence-357199-        8508 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mouse full length TGF-beta-2 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8508)
  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 8508)
  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 TGFb2
FEATURES             Location/Qualifiers
     source          1..8508
                     /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    4579..4634
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4633..4652)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(5344..5932)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5424..5443)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6018..6827)
                     /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     6741..6760
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(6846..6893)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             complement(7319..7918)
                     /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(7402..7422)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     7899..7918
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(7928..8427)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7976..7995)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     8369..8392
                     /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 aggkgcggcg 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 cgagctgtct acctgcagca ccctcgacat ggatcagttt atgcgcaaga
     1861 ggatcgaggc catccgcggg cagatcctga gcaagctgaa gctcaccagc cccccggaag
     1921 actatccgga gccggatgag gtccccccgg aggtgatttc catctacaac agtaccaggg
     1981 acttactgca ggagaaggca agccggaggg cagccgcctg cgagcgcgag cggagcgacg
     2041 aggagtacta cgccaaggag gtttataaaa tcgacatgcc gtcccacctc ccctccgaaa
     2101 atgccatccc gcccactttc tacagaccct acttcagaat cgtccgcttt gatgtctcaa
     2161 caatggagaa aaatgcttcg aatctggtga aggcagagtt cagggtcttc cgcttgcaaa
     2221 accccaaagc cagagtggcc gagcagcgga ttgaactgta tcagatcctt aaatccaaag
     2281 acttaacatc tcccacccag cgctacatcg atagcaaggt tgtgaaaacc agagcggagg
     2341 gtgaatggct ctccttcgac gtgacagacg ctgtgcagga gtggcttcac cacaaagaca
     2401 ggaacctggg gtttaaaata agtttacact gcccctgctg taccttcgtg ccgtctaata
     2461 attacatcat cccgaataaa agcgaagagc tcgaggcgag atttgcaggt attgatggca
     2521 cctctacata tgccagtggt gatcagaaaa ctataaagtc cactaggaaa aaaaccagtg
     2581 ggaagacccc acatctcctg ctaatgttgt tgccctccta cagactggag tcacaacagt
     2641 ccagccggcg gaagaagcgc gctttggatg ctgcctactg ctttagaaat gtgcaggata
     2701 attgctgcct tcgccctctt tacattgatt ttaagaggga tcttggatgg aaatggatcc
     2761 atgaacccaa agggtacaat gctaacttct gtgctggggc atgcccatat ctatggagtt
     2821 cagacactca acacaccaaa gtcctcagcc tgtacaacac cataaatccc gaagcttccg
     2881 cttccccttg ctgtgtgtcc caggatctgg aaccactgac cattctctat tacattggaa
     2941 atacgcccaa gatcgaacag ctttccaata tgattgtcaa gtcttgtaaa tgcagctaat
     3001 gagctagcgg ttgaggtctc taaaagcgtc ttcctgttct catcacatca tatcaaggtt
     3061 atataccatc aatattgcca cagatgttac ttagcctttt aatatttctc taatttagtg
     3121 tatatgcaat gatagttctc tgatttctga gattgagttt ctcatgtgta atgattattt
     3181 agagtttctc tttcatctgt tcaaattttt gtctagtttt attttttact gatttgtaag
     3241 acttcttttt ataatctgca tattacaatt ctctttactg gggtgttgca aatattttct
     3301 gtcattctat ggcctgactt ttcttaatgg ttttttaatt ttaaaaataa gtcttaatat
     3361 tcatgcaatc taattaacaa tcttttcttt gtggttagga ctttgagtca taagaaattt
     3421 ttctctacac tgaagtcatg atggcatgct tctatattat tttctaaaag atttaaagtt
     3481 ttgccttctc catttagact tataattcac tggaattttt ttgtgtgtat ggtatgacat
     3541 atgggttccc ttttattttt tacatataaa tatatttccc tgtttttcta aaaaagaaaa
     3601 agatcatcat tttcccattg taaaatgcca tatttttttc ataggtcact tacatatatc
     3661 aatgggtctg tttctgagct ctactctatt ttatcagcct cactgtctat ccccacacat
     3721 ctcatgcttt gctctaaatc ttgatattta gtggaacatt ctttcccatt ttgttctaca
     3781 agaatatttt tgttattgtc tttgggcttt ctatatacat tttgaaatga ggttgacaag
     3841 ttaataacag gcctattgat tggaaagttt gtcaacgaat tgtgggtctt ttggggtttg
     3901 ctgccccttt tacgcaatgt ggatatcctg ctttaatgcc tttatatgca tgtatacaag
     3961 caaaacaggc ttttactttc tcgccaactt acaaggcctt tctcagtaaa cagtatatga
     4021 ccctttaccc cgttgctcgg caacggcctg gtctgtgcca agtgtttgct gacgcaaccc
     4081 ccactggttg gggcttggcc ataggccatc agcgcatgcg tggaaccttt gtgtctcctc
     4141 tgccgatcca tactgcggaa ctcctagccg cttgttttgc tcgcagcagg tctggagcaa
     4201 acctcatcgg gaccgacaat tctgtcgtac tctcccgcaa gtatacatcg tttccatggc
     4261 tgctaggctg tgctgccaac tggatcctgc gcgggacgtc ctttgtttac gtcccgtcgg
     4321 cgctgaatcc cgcggacgac ccctcccggg gccgcttggg gctctaccgc ccgcttctcc
     4381 gtctgccgta ccgtccgacc acggggcgca cctctcttta cgcggactcc ccgtctgtgc
     4441 cttctcatct gccggaccgt gtgcacttcg cttcacctct gcacgtcgca tggaggccac
     4501 cgtgaacgcc caccggaacc tgcccaaggt cttgcataag aggactcttg gactttcagc
     4561 aatgtcatct cagtggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt
     4621 ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg
     4681 agtatttggt ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt
     4741 ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata
     4801 gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt
     4861 taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac
     4921 tactcccagt catagctgtc cctcttctct tatggagatc tttgcatttt tagacattta
     4981 gaagcctata tcttgttaca gaattggaat tacacaaaaa ttctaccata ttttgaaagc
     5041 ttaggttgtt ctgaaaaaaa caatatattg ttttcctggg taaactaaaa gtcccctcga
     5101 ggaaaggccc ctaaagtgaa acagtgcaaa acgttcaaaa actgtctggc aatacaagtt
     5161 ccactttgac caaaacggct ggcagtaaaa gggttaagaa gactgtcagc cttgagcggt
     5221 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5281 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5341 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5401 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5461 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5521 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5581 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5641 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5701 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5761 ggctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5821 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5881 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5941 tttgatcttt tctacggggt ctgacgctca gtggaacgac gcgcgcgtaa ctcacgttaa
     6001 gggattttgg tcatgagtta gaaaaactca tcgagcatca aatgaaactg caatttattc
     6061 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
     6121 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt
     6181 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
     6241 tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag
     6301 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
     6361 ttattcattc gtgattgcgc ctgagcgagg cgaaatacgc gatcgctgtt aaaaggacaa
     6421 ttacaaacag gaatcgagtg caaccggcgc aggaacactg ccagcgcatc aacaatattt
     6481 tcacctgaat caggatattc ttctaatacc tggaacgctg tttttccggg gatcgcagtg
     6541 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagtggcata
     6601 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct
     6661 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaagcg atagattgtc
     6721 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg
     6781 ttggaattta atcgcggcct cgacgtttcc cgttggatat ggctcatttt ttacttcctc
     6841 accttgtcgt attatactat gccgatatac tatgccgatg attaattgtc gacactgcgg
     6901 gggctctgtg tggtaagcag gtcttaacct ttttactgcc aatgacgcat gggatacgtc
     6961 gtggcagtaa aagggcttaa atgccaacga cgcgtcccat acgttgttgg cattttaatt
     7021 cttctctctg cagcggcagc atgtgccgcc gctgcagaga gtttctagcg atgacagccc
     7081 ctctgggcaa cgagccgggg gggctgtctt tctttatgtt ttaaatgcac tgacctccca
     7141 cattcccttt ttagtaaaat attcagaaat aatttaaata catcattgca atgaaaataa
     7201 atgtttttta ttaggcagaa tccagatgct caaggccctt cataatatcc cccagtttag
     7261 tagttggact tagggaacaa aggaaccttt aatagaaatt ggacagcaag aaagcgagtc
     7321 aggcaccggg cttgcgggtc atgcaccagg tgcgcggtcc ttcgggcacc tcgacgtcgg
     7381 cggtgacggt gaagccgagc cgctcgtaga aggggaggtt gcggggcgcg gatgtctcca
     7441 ggaaggcggg caccccggcg cgctcggccg cctccactcc ggggagcacg acggcgctgc
     7501 ccagaccctt gccctggtgg tcgggcgaca cgccgacggt ggccaggaac cacgcgggct
     7561 ccttgggccg gtgcggcgcc aggaggcctt ccatctgttg ctgcgcggcc agccgggaac
     7621 cgctcaactc ggccatgcgc gggccgatct cggcgaacac cgcccccgct tcgacgctct
     7681 ccggcgtggt ccagaccgcc accgcggcgc cgtcgtccgc gacccacacc ttgccgatgt
     7741 cgagcccgac gcgcgtgagg aagagttctt gcagctcggt gacccgctcg atgtggcggt
     7801 ccggatcgac ggtgtggcgc gtggcggggt agtcggcgaa cgcggcggcg agggtgcgta
     7861 cggccctggg gacgtcgtcg cgggtggcga ggcgcaccgt gggcttgtac tcggtcatgg
     7921 tggcggacga aaggcccgga gatgaggaag aggagaacag cgcggcagac gtgcgctttt
     7981 gaagcgtgca gaatgccggg cctccggagg accttcgggc gcccgccccg cccctgagcc
     8041 cgcccctgag cccgcccccg gacccacccc ttcccagcct ctgagcccag aaagcgaagg
     8101 agcaaagctg ctattggccg ctgccccaaa ggcctacccg cttccattgc tcagcggtgc
     8161 tgtccatctg cacgagacta gtgagtcgtg ctacttccat ttgtcacgtc ctgcacgacg
     8221 cgagctgcgg ggcggggggg aacttcctga ctaggggagg agtagaaggt ggcgcgaagg
     8281 ggccaccaaa gaacggagcc ggttggcgcc taccggtgga tgtggaatgt gtgcgaggcc
     8341 agaggccact tgtgtagcgc caagtgccca gcggggctgc taaagcgcat gctccagact
     8401 gccttgggaa aagcgcctcc cctacccggt agagaaactt gatctgtcgc cgcaattcaa
     8461 acttcgtgag gctccggtgc ccgtcagtga cctgctatac tctggaga
//
