LOCUS       sequence-357198-        8412 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mouse full length TGF-beta-1 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8412)
  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 8412)
  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 TGFb1
FEATURES             Location/Qualifiers
     source          1..8412
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        3..306
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     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    4483..4538
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4537..4556)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(5248..5836)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5328..5347)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5922..6731)
                     /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     6645..6664
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(6750..6797)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             complement(7223..7822)
                     /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(7306..7326)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     7803..7822
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(7832..8331)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7880..7899)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     8273..8296
                     /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 sgrggsggrg
      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 cgagctctcc acctgcaaga ccatcgacat ggagctggtg aaacggaagc
     1861 gcatcgaagc catccgtggc cagatcctgt ccaaactaag gctcgccagt cccccaagcc
     1921 agggggaggt accgcccggc ccgctgcccg aggcggtgct cgctttgtac aacagcaccc
     1981 gcgaccgggt ggcaggcgag agcgccgacc cagagccgga gcccgaagcg gactactatg
     2041 ctaaagaggt cacccgcgtg ctaatggtgg accgcaacaa cgccatctat gagaaaacca
     2101 aagacatctc acacagtata tatatgttct tcaatacgtc agacattcgg gaagcagtgc
     2161 ccgaaccccc attgctgtcc cgtgcagagc tgcgcttgca gagattaaaa tcaagtgtgg
     2221 agcaacatgt ggaactctac cagaaatata gcaacaattc ctggcgttac cttggtaacc
     2281 ggctgctgac ccccactgat acgcctgagt ggctgtcttt tgacgtcact ggagttgtac
     2341 ggcagtggct gaaccaagga gacggaatac agggctttcg attcagcgct cactgctctt
     2401 gtgacagcaa agataacaaa ctccacgtgg aaatcaacgg gatcagcccc aaacgtcggg
     2461 gcgacctggg caccatccat gacatgaacc ggcccttcct gctcctcatg gccacccccc
     2521 tggaaagggc ccagcacctg cacagctcac ggcaccggag agccctggat accaactatt
     2581 gcttcagctc cacagagaag aactgctgtg tgcggcagct gtacattgac tttaggaagg
     2641 acctgggttg gaagtggatc cacgagccca agggctacca tgccaacttc tgtctgggac
     2701 cctgccccta tatttggagc ctggacacac agtacagcaa ggtccttgcc ctctacaacc
     2761 aacacaaccc gggcgcttcg gcgtcaccgt gctgcgtgcc gcaggctttg gagccactgc
     2821 ccatcgtcta ctacgtgggt cgcaagccca aggtggagca gttgtccaac atgattgtgc
     2881 gctcctgcaa gtgcagctaa tgataagcta gcggttgagg tctctaaaag cgtcttcctg
     2941 ttctcatcac atcatatcaa ggttatatac catcaatatt gccacagatg ttacttagcc
     3001 ttttaatatt tctctaattt agtgtatatg caatgatagt tctctgattt ctgagattga
     3061 gtttctcatg tgtaatgatt atttagagtt tctctttcat ctgttcaaat ttttgtctag
     3121 ttttattttt tactgatttg taagacttct ttttataatc tgcatattac aattctcttt
     3181 actggggtgt tgcaaatatt ttctgtcatt ctatggcctg acttttctta atggtttttt
     3241 aattttaaaa ataagtctta atattcatgc aatctaatta acaatctttt ctttgtggtt
     3301 aggactttga gtcataagaa atttttctct acactgaagt catgatggca tgcttctata
     3361 ttattttcta aaagatttaa agttttgcct tctccattta gacttataat tcactggaat
     3421 ttttttgtgt gtatggtatg acatatgggt tcccttttat tttttacata taaatatatt
     3481 tccctgtttt tctaaaaaag aaaaagatca tcattttccc attgtaaaat gccatatttt
     3541 tttcataggt cacttacata tatcaatggg tctgtttctg agctctactc tattttatca
     3601 gcctcactgt ctatccccac acatctcatg ctttgctcta aatcttgata tttagtggaa
     3661 cattctttcc cattttgttc tacaagaata tttttgttat tgtctttggg ctttctatat
     3721 acattttgaa atgaggttga caagttaata acaggcctat tgattggaaa gtttgtcaac
     3781 gaattgtggg tcttttgggg tttgctgccc cttttacgca atgtggatat cctgctttaa
     3841 tgcctttata tgcatgtata caagcaaaac aggcttttac tttctcgcca acttacaagg
     3901 cctttctcag taaacagtat atgacccttt accccgttgc tcggcaacgg cctggtctgt
     3961 gccaagtgtt tgctgacgca acccccactg gttggggctt ggccataggc catcagcgca
     4021 tgcgtggaac ctttgtgtct cctctgccga tccatactgc ggaactccta gccgcttgtt
     4081 ttgctcgcag caggtctgga gcaaacctca tcgggaccga caattctgtc gtactctccc
     4141 gcaagtatac atcgtttcca tggctgctag gctgtgctgc caactggatc ctgcgcggga
     4201 cgtcctttgt ttacgtcccg tcggcgctga atcccgcgga cgacccctcc cggggccgct
     4261 tggggctcta ccgcccgctt ctccgtctgc cgtaccgtcc gaccacgggg cgcacctctc
     4321 tttacgcgga ctccccgtct gtgccttctc atctgccgga ccgtgtgcac ttcgcttcac
     4381 ctctgcacgt cgcatggagg ccaccgtgaa cgcccaccgg aacctgccca aggtcttgca
     4441 taagaggact cttggacttt cagcaatgtc atctcagtgg ctaataaagg aaatttattt
     4501 tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg
     4561 caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata
     4621 tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc
     4681 ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat
     4741 tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag
     4801 ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgga
     4861 gatctttgca tttttagaca tttagaagcc tatatcttgt tacagaattg gaattacaca
     4921 aaaattctac catattttga aagcttaggt tgttctgaaa aaaacaatat attgttttcc
     4981 tgggtaaact aaaagtcccc tcgaggaaag gcccctaaag tgaaacagtg caaaacgttc
     5041 aaaaactgtc tggcaataca agttccactt tgaccaaaac ggctggcagt aaaagggtta
     5101 agaagactgt cagccttgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5161 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5221 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5281 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5341 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5401 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5461 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5521 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5581 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5641 tgctacagag ttcttgaagt ggtgggctaa ctacggctac actagaagaa cagtatttgg
     5701 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5761 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5821 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5881 cgacgcgcgc gtaactcacg ttaagggatt ttggtcatga gttagaaaaa ctcatcgagc
     5941 atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc
     6001 cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg
     6061 tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca
     6121 aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc
     6181 aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca
     6241 aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc gaggcgaaat
     6301 acgcgatcgc tgttaaaagg acaattacaa acaggaatcg agtgcaaccg gcgcaggaac
     6361 actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa tacctggaac
     6421 gctgtttttc cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa
     6481 tgcttgatgg tcggaagtgg cataaattcc gtcagccagt ttagtctgac catctcatct
     6541 gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc
     6601 ttcccataca agcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta
     6661 tacccatata aatcagcatc catgttggaa tttaatcgcg gcctcgacgt ttcccgttgg
     6721 atatggctca ttttttactt cctcaccttg tcgtattata ctatgccgat atactatgcc
     6781 gatgattaat tgtcgacact gcgggggctc tgtgtggtaa gcaggtctta acctttttac
     6841 tgccaatgac gcatgggata cgtcgtggca gtaaaagggc ttaaatgcca acgacgcgtc
     6901 ccatacgttg ttggcatttt aattcttctc tctgcagcgg cagcatgtgc cgccgctgca
     6961 gagagtttct agcgatgaca gcccctctgg gcaacgagcc gggggggctg tctttcttta
     7021 tgttttaaat gcactgacct cccacattcc ctttttagta aaatattcag aaataattta
     7081 aatacatcat tgcaatgaaa ataaatgttt tttattaggc agaatccaga tgctcaaggc
     7141 ccttcataat atcccccagt ttagtagttg gacttaggga acaaaggaac ctttaataga
     7201 aattggacag caagaaagcg agtcaggcac cgggcttgcg ggtcatgcac caggtgcgcg
     7261 gtccttcggg cacctcgacg tcggcggtga cggtgaagcc gagccgctcg tagaagggga
     7321 ggttgcgggg cgcggatgtc tccaggaagg cgggcacccc ggcgcgctcg gccgcctcca
     7381 ctccggggag cacgacggcg ctgcccagac ccttgccctg gtggtcgggc gacacgccga
     7441 cggtggccag gaaccacgcg ggctccttgg gccggtgcgg cgccaggagg ccttccatct
     7501 gttgctgcgc ggccagccgg gaaccgctca actcggccat gcgcgggccg atctcggcga
     7561 acaccgcccc cgcttcgacg ctctccggcg tggtccagac cgccaccgcg gcgccgtcgt
     7621 ccgcgaccca caccttgccg atgtcgagcc cgacgcgcgt gaggaagagt tcttgcagct
     7681 cggtgacccg ctcgatgtgg cggtccggat cgacggtgtg gcgcgtggcg gggtagtcgg
     7741 cgaacgcggc ggcgagggtg cgtacggccc tggggacgtc gtcgcgggtg gcgaggcgca
     7801 ccgtgggctt gtactcggtc atggtggcgg acgaaaggcc cggagatgag gaagaggaga
     7861 acagcgcggc agacgtgcgc ttttgaagcg tgcagaatgc cgggcctccg gaggaccttc
     7921 gggcgcccgc cccgcccctg agcccgcccc tgagcccgcc cccggaccca ccccttccca
     7981 gcctctgagc ccagaaagcg aaggagcaaa gctgctattg gccgctgccc caaaggccta
     8041 cccgcttcca ttgctcagcg gtgctgtcca tctgcacgag actagtgagt cgtgctactt
     8101 ccatttgtca cgtcctgcac gacgcgagct gcggggcggg ggggaacttc ctgactaggg
     8161 gaggagtaga aggtggcgcg aaggggccac caaagaacgg agccggttgg cgcctaccgg
     8221 tggatgtgga atgtgtgcga ggccagaggc cacttgtgta gcgccaagtg cccagcgggg
     8281 ctgctaaagc gcatgctcca gactgccttg ggaaaagcgc ctcccctacc cggtagagaa
     8341 acttgatctg tcgccgcaat tcaaacttcg tgaggctccg gtgcccgtca gtgacctgct
     8401 atactctgga ga
//
