LOCUS       sequence-469707-        4454 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway ORF Entry clone of mouse Tgfb2 with stop codon (for native 
            or N-terminal fusions); C226S/C228S/C229S mutations.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4454)
  TITLE     Esposito lab plasmids 2025
REFERENCE   2  (bases 1 to 4454)
  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"
COMMENT     Sequence Label: 8522-E05
FEATURES             Location/Qualifiers
     source          1..4454
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             668..688
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(1935..2034)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2051..2070)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2052..2070)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2075..2091)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2075..2091)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2522..3313
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     oriT            3435..3544
                     /note="incP origin of transfer"
     rep_origin      3804..4392
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4293..4312
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agttggcgaa aacctgtact tccaaggcac catgcactac tgtgtgctga gcaccttttt
      721 gctcctgcat ctggtcccgg tggcgctcag tctgtctacc tgcagcaccc tcgacatgga
      781 tcagtttatg cgcaagagga tcgaggccat ccgcgggcag atcctgagca agctgaagct
      841 caccagcccc ccggaagact atccggagcc ggatgaggtc cccccggagg tgatttccat
      901 ctacaacagt accagggact tactgcagga gaaggcaagc cggagggcag ccgcctgcga
      961 gcgcgagcgg agcgacgagg agtactacgc caaggaggtt tataaaatcg acatgccgtc
     1021 ccacctcccc tccgaaaatg ccatcccgcc cactttctac agaccctact tcagaatcgt
     1081 ccgctttgat gtctcaacaa tggagaaaaa tgcttcgaat ctggtgaagg cagagttcag
     1141 ggtcttccgc ttgcaaaacc ccaaagccag agtggccgag cagcggattg aactgtatca
     1201 gatccttaaa tccaaagact taacatctcc cacccagcgc tacatcgata gcaaggttgt
     1261 gaaaaccaga gcggagggtg aatggctctc cttcgacgtg acagacgctg tgcaggagtg
     1321 gcttcaccac aaagacagga acctggggtt taaaataagt ttacactccc cctcctctac
     1381 cttcgtgccg tctaataatt acatcatccc gaataaaagc gaagagctcg aggcgagatt
     1441 tgcaggtatt gatggcacct ctacatatgc cagtggtgat cagaaaacta taaagtccac
     1501 taggaaaaaa accagtggga agaccccaca tctcctgcta atgttgttgc cctcctacag
     1561 actggagtca caacagtcca gccggcggaa gaagcgcgct ttggatgctg cctactgctt
     1621 tagaaatgtg caggataatt gctgccttcg ccctctttac attgatttta agagggatct
     1681 tggatggaaa tggatccatg aacccaaagg gtacaatgct aacttctgtg ctggggcatg
     1741 cccatatcta tggagttcag acactcaaca caccaaagtc ctcagcctgt acaacaccat
     1801 aaatcccgaa gcttccgctt ccccttgctg tgtgtcccag gatctggaac cactgaccat
     1861 tctctattac attggaaata cgcccaagat cgaacagctt tccaatatga ttgtcaagtc
     1921 ttgtaaatgc agctaatcaa ctttcttgta caaagttggc attataagaa agcattgctt
     1981 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     2041 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     2101 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgcct
     2161 cctctagacc agccaggaca gaaatgcctc gacttcgctg ctacccaagg ttgccgggtg
     2221 acgcacaccg tggaaacgga tgaaggcacg aacccagtgg acataagcct gttcggttcg
     2281 taagctgtaa tgcaagtagc gtatgcgctc acgcaactgg tccagaacct tgaccgaacg
     2341 cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttggggt
     2401 acagtctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat
     2461 gttatggagc agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaaacat
     2521 tatgagggaa gcggtgatcg ccgaagtatc gactcaacta tcagaggtag ttggcgtcat
     2581 cgagcgccat ctcgaaccga cgttgctggc cgtacatttg tacggctccg cagtggatgg
     2641 cggcctgaag ccacacagtg atattgattt gctggttacg gtgaccgtaa ggcttgatga
     2701 aacaacgcgg cgagctttga tcaacgacct tttggaaact tcggcttccc ctggagagag
     2761 cgagattctc cgcgctgtag aagtcaccat tgttgtgcac gacgacatca ttccgtggcg
     2821 ttatccagct aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg
     2881 tatcttcgag ccagccacga tcgacattga tctggctatc ttgctgacaa aagcaagaga
     2941 acatagcgtt gccttggtag gtccagcggc ggaggaactc tttgatccgg ttcctgaaca
     3001 ggatctattt gaggcgctaa atgaaacctt aacgctatgg aactcgccgc ccgactgggc
     3061 tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg
     3121 caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc cggcccagta
     3181 tcagcccgtc atacttgaag ctagacaggc ttatcttgga caagaagaag atcgcttggc
     3241 ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt
     3301 agtcggcaaa taaccctcga ctctagagga tccccgggta ccgagctcga attccgtgat
     3361 aggtgggctg cccttcctgg ttggcttggt ttcatcagcc atccgcttgc cctcatctgt
     3421 tacgccggcg gtagccggcc agcctcgcag agcaggattc ccgttgagca ccgccaggtg
     3481 cgaataaggg acagtgaaga aggaacaccc gctcgcgggt gggcctactt cacctatcct
     3541 gcccggctga cgccgttgga tacaccaagg aaagtctaca cgaacccttt ggcaaaatcc
     3601 tgtatatcgt gcgaaaaagg atggatatac cgaaaaaatc gctataatga ccccgaagca
     3661 gggttatgca gcggaaaacg gaattcgagc tcggtacccg gggatcctct agagtcgagc
     3721 cacccatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
     3781 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3841 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3901 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     3961 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4021 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4081 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4141 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4201 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4261 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4321 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4381 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4441 tttgctcaca tgtt
//
