LOCUS       sequence-468853-        4487 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway ORF Entry clone of human SMAD3 with stop codon (for native 
            or N-terminal fusions); T179V/S204A/S208A/S213A mutations (All four 
            linker sites dead).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4487)
  TITLE     Esposito lab plasmids 2025
REFERENCE   2  (bases 1 to 4487)
  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-E13
FEATURES             Location/Qualifiers
     source          1..4487
                     /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(1968..2067)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2084..2103)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2085..2103)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2108..2124)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2108..2124)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2555..3346
                     /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            3468..3577
                     /note="incP origin of transfer"
     rep_origin      3837..4425
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4326..4345
                     /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 catgtcgtcc atcctgcctt tcactccccc
      721 gatcgtgaag cgcctgctgg gctggaagaa gggcgagcag aacgggcagg aggagaaatg
      781 gtgcgagaag gcggtcaaga gcctggtcaa gaaactcaag aagacggggc agctggacga
      841 gctggagaag gccatcacca cgcagaacgt caacaccaag tgcatcacca tccccaggtc
      901 cctggatggc cggttgcagg tgtcccatcg gaaggggctc cctcatgtca tctactgccg
      961 cctgtggcga tggccagacc tgcacagcca ccacgagctg cgggccatgg agctgtgtga
     1021 gttcgccttc aatatgaaga aggacgaggt ctgcgtgaat ccctaccact accagagagt
     1081 agagacacca gttctacctc ctgtgttggt gccacgccac acagagatcc cggccgagtt
     1141 ccccccactg gacgactaca gccattccat ccccgaaaac actaacttcc ccgcaggcat
     1201 cgagccccag agcaatattc cagaggtccc accccctggc tacctgagtg aagatggaga
     1261 aaccagtgac caccagatga accacagcat ggacgcaggt gctccaaacc tagccccgaa
     1321 tccgatggcc ccagcacata ataacttgga cctgcagcca gttacctact gcgagccggc
     1381 cttctggtgc tccatctcct actacgagct gaaccagcgc gtcggggaga cattccacgc
     1441 ctcgcagcca tccatgactg tggatggctt caccgacccc tccaattcgg agcgcttctg
     1501 cctagggctg ctctccaatg tcaacaggaa tgcagcagtg gagctgacac ggagacacat
     1561 cggaagaggc gtgcggctct actacatcgg aggggaggtc ttcgcagagt gcctcagtga
     1621 cagcgctatt tttgtccagt ctcccaactg taaccagcgc tatggctggc acccggccac
     1681 cgtctgcaag atcccaccag gatgcaacct gaagatcttc aacaaccagg agttcgctgc
     1741 cctcctggcc cagtcggtca accagggctt tgaggctgtc taccagttga cccgaatgtg
     1801 caccatccgc atgagcttcg tcaaaggctg gggagcggag tacaggagac agactgtgac
     1861 cagtaccccc tgctggattg agctgcacct gaatgggcct ttgcagtggc ttgacaaggt
     1921 cctcacccag atgggctccc caagcatccg ctgttccagt gtgtcttaat caactttctt
     1981 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     2041 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     2101 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     2161 cattgcacaa gataaaaata tatcatcatg cctcctctag accagccagg acagaaatgc
     2221 ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac ggatgaaggc
     2281 acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg
     2341 ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg cagtggcggt
     2401 tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg catccaagca
     2461 gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg atgttacgca
     2521 gcagggcagt cgccctaaaa caaagttaaa cattatgagg gaagcggtga tcgccgaagt
     2581 atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac cgacgttgct
     2641 ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca gtgatattga
     2701 tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt tgatcaacga
     2761 ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg tagaagtcac
     2821 cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg aactgcaatt
     2881 tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca cgatcgacat
     2941 tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg taggtccagc
     3001 ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc taaatgaaac
     3061 cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg tagtgcttac
     3121 gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc
     3181 cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg aagctagaca
     3241 ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt
     3301 tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct cgactctaga
     3361 ggatccccgg gtaccgagct cgaattccgt gataggtggg ctgcccttcc tggttggctt
     3421 ggtttcatca gccatccgct tgccctcatc tgttacgccg gcggtagccg gccagcctcg
     3481 cagagcagga ttcccgttga gcaccgccag gtgcgaataa gggacagtga agaaggaaca
     3541 cccgctcgcg ggtgggccta cttcacctat cctgcccggc tgacgccgtt ggatacacca
     3601 aggaaagtct acacgaaccc tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata
     3661 taccgaaaaa atcgctataa tgaccccgaa gcagggttat gcagcggaaa acggaattcg
     3721 agctcggtac ccggggatcc tctagagtcg agccacccat gaccaaaatc ccttaacgtg
     3781 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3841 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3901 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3961 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     4021 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4081 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4141 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4201 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4261 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4321 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4381 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4441 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
