LOCUS       sequence-468878-        4382 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway ORF Entry clone of mouse Tgfb1 with stop codon (for native 
            or N-terminal fusions); C223S/C225S mutations.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4382)
  TITLE     Esposito lab plasmids 2025
REFERENCE   2  (bases 1 to 4382)
  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-E04
FEATURES             Location/Qualifiers
     source          1..4382
                     /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(1863..1962)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1979..1998)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1980..1998)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2003..2019)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2003..2019)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2450..3241
                     /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            3363..3472
                     /note="incP origin of transfer"
     rep_origin      3732..4320
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4221..4240
                     /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 catgccgccc tcggggctgc ggctactgcc
      721 gcttctgctc ccactcccgt ggcttctagt gctgacgccc gggaggccag ccgcgggact
      781 ctccacctgc aagaccatcg acatggagct ggtgaaacgg aagcgcatcg aagccatccg
      841 tggccagatc ctgtccaaac taaggctcgc cagtccccca agccaggggg aggtaccgcc
      901 cggcccgctg cccgaggcgg tgctcgcttt gtacaacagc acccgcgacc gggtggcagg
      961 cgagagcgcc gacccagagc cggagcccga agcggactac tatgctaaag aggtcacccg
     1021 cgtgctaatg gtggaccgca acaacgccat ctatgagaaa accaaagaca tctcacacag
     1081 tatatatatg ttcttcaata cgtcagacat tcgggaagca gtgcccgaac ccccattgct
     1141 gtcccgtgca gagctgcgct tgcagagatt aaaatcaagt gtggagcaac atgtggaact
     1201 ctaccagaaa tatagcaaca attcctggcg ttaccttggt aaccggctgc tgacccccac
     1261 tgatacgcct gagtggctgt cttttgacgt cactggagtt gtacggcagt ggctgaacca
     1321 aggagacgga atacagggct ttcgattcag cgctcactcc tcttctgaca gcaaagataa
     1381 caaactccac gtggaaatca acgggatcag ccccaaacgt cggggcgacc tgggcaccat
     1441 ccatgacatg aaccggccct tcctgctcct catggccacc cccctggaaa gggcccagca
     1501 cctgcacagc tcacggcacc ggagagccct ggataccaac tattgcttca gctccacaga
     1561 gaagaactgc tgtgtgcggc agctgtacat tgactttagg aaggacctgg gttggaagtg
     1621 gatccacgag cccaagggct accatgccaa cttctgtctg ggaccctgcc cctatatttg
     1681 gagcctggac acacagtaca gcaaggtcct tgccctctac aaccaacaca acccgggcgc
     1741 ttcggcgtca ccgtgctgcg tgccgcaggc tttggagcca ctgcccatcg tctactacgt
     1801 gggtcgcaag cccaaggtgg agcagttgtc caacatgatt gtgcgctcct gcaagtgcag
     1861 ctaatcaact ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg
     1921 caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc
     1981 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca
     2041 aaatctctga tgttacattg cacaagataa aaatatatca tcatgcctcc tctagaccag
     2101 ccaggacaga aatgcctcga cttcgctgct acccaaggtt gccgggtgac gcacaccgtg
     2161 gaaacggatg aaggcacgaa cccagtggac ataagcctgt tcggttcgta agctgtaatg
     2221 caagtagcgt atgcgctcac gcaactggtc cagaaccttg accgaacgca gcggtggtaa
     2281 cggcgcagtg gcggttttca tggcttgtta tgactgtttt tttggggtac agtctatgcc
     2341 tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag
     2401 caacgatgtt acgcagcagg gcagtcgccc taaaacaaag ttaaacatta tgagggaagc
     2461 ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg agcgccatct
     2521 cgaaccgacg ttgctggccg tacatttgta cggctccgca gtggatggcg gcctgaagcc
     2581 acacagtgat attgatttgc tggttacggt gaccgtaagg cttgatgaaa caacgcggcg
     2641 agctttgatc aacgaccttt tggaaacttc ggcttcccct ggagagagcg agattctccg
     2701 cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa
     2761 gcgcgaactg caatttggag aatggcagcg caatgacatt cttgcaggta tcttcgagcc
     2821 agccacgatc gacattgatc tggctatctt gctgacaaaa gcaagagaac atagcgttgc
     2881 cttggtaggt ccagcggcgg aggaactctt tgatccggtt cctgaacagg atctatttga
     2941 ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc gactgggctg gcgatgagcg
     3001 aaatgtagtg cttacgttgt cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc
     3061 gaaggatgtc gctgccgact gggcaatgga gcgcctgccg gcccagtatc agcccgtcat
     3121 acttgaagct agacaggctt atcttggaca agaagaagat cgcttggcct cgcgcgcaga
     3181 tcagttggaa gaatttgtcc actacgtgaa aggcgagatc accaaggtag tcggcaaata
     3241 accctcgact ctagaggatc cccgggtacc gagctcgaat tccgtgatag gtgggctgcc
     3301 cttcctggtt ggcttggttt catcagccat ccgcttgccc tcatctgtta cgccggcggt
     3361 agccggccag cctcgcagag caggattccc gttgagcacc gccaggtgcg aataagggac
     3421 agtgaagaag gaacacccgc tcgcgggtgg gcctacttca cctatcctgc ccggctgacg
     3481 ccgttggata caccaaggaa agtctacacg aaccctttgg caaaatcctg tatatcgtgc
     3541 gaaaaaggat ggatataccg aaaaaatcgc tataatgacc ccgaagcagg gttatgcagc
     3601 ggaaaacgga attcgagctc ggtacccggg gatcctctag agtcgagcca cccatgacca
     3661 aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc
     3721 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3781 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3841 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     3901 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     3961 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4021 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4081 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4141 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4201 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4261 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4321 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     4381 tt
//
