LOCUS       sequence-468850-        4613 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway ORF Entry clone of human SMAD2 with stop codon (for native 
            or N-terminal fusions); S464A/S465A/S467A mutations (C-terminal 
            phosphorylation sites dead).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4613)
  TITLE     Esposito lab plasmids 2025
REFERENCE   2  (bases 1 to 4613)
  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-E10
FEATURES             Location/Qualifiers
     source          1..4613
                     /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(2094..2193)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2210..2229)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2211..2229)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2234..2250)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2234..2250)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2681..3472
                     /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            3594..3703
                     /note="incP origin of transfer"
     rep_origin      3963..4551
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4452..4471
                     /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 atcttgccat tcacgccgcc
      721 agttgtgaag agactgctgg gatggaagaa gtcagctggt gggtctggag gagcaggcgg
      781 aggagagcag aatgggcagg aagaaaagtg gtgtgagaaa gcagtgaaaa gtctggtgaa
      841 gaagctaaag aaaacaggac gattagatga gcttgagaaa gccatcacca ctcaaaactg
      901 taatactaaa tgtgttacca taccaagcac ttgctctgaa atttggggac tgagtacacc
      961 aaatacgata gatcagtggg atacaacagg cctttacagc ttctctgaac aaaccaggtc
     1021 tcttgatggt cgtctccagg tatcccatcg aaaaggattg ccacatgtta tatattgccg
     1081 attatggcgc tggcctgatc ttcacagtca tcatgaactc aaggcaattg aaaactgcga
     1141 atatgctttt aatcttaaaa aggatgaagt atgtgtaaac ccttaccact atcagagagt
     1201 tgagacacca gttttgcctc cagtattagt gccccgacac accgagatcc taacagaact
     1261 tccgcctctg gatgactata ctcactccat tccagaaaac actaacttcc cagcaggaat
     1321 tgagccacag agtaattata ttccagaaac gccacctcct ggatatatca gtgaagatgg
     1381 agaaacaagt gaccaacagt tgaatcaaag tatggacaca ggctctccag cagaactatc
     1441 tcctactact ctttcccctg ttaatcatag cttggattta cagccagtta cttactcaga
     1501 acctgcattt tggtgttcga tagcatatta tgaattaaat cagagggttg gagaaacctt
     1561 ccatgcatca cagccctcac tcactgtaga tggctttaca gacccatcaa attcagagag
     1621 gttctgctta ggtttactct ccaatgttaa ccgaaatgcc acggtagaaa tgacaagaag
     1681 gcatatagga agaggagtgc gcttatacta cataggtggg gaagtttttg ctgagtgcct
     1741 aagtgatagt gcaatctttg tgcagagccc caattgtaat cagagatatg gctggcaccc
     1801 tgcaacagtg tgtaaaattc caccaggctg taatctgaag atcttcaaca accaggaatt
     1861 tgctgctctt ctggctcagt ctgttaatca gggttttgaa gccgtctatc agctaactag
     1921 aatgtgcacc ataagaatga gttttgtgaa agggtgggga gcagaatacc gaaggcagac
     1981 ggtaacaagt actccttgct ggattgaact tcatctgaat ggacctctac agtggttgga
     2041 caaagtatta actcagatgg gatccccttc agtgcgttgc gcagccatgg cataatcaac
     2101 tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca
     2161 ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga
     2221 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     2281 atgttacatt gcacaagata aaaatatatc atcatgcctc ctctagacca gccaggacag
     2341 aaatgcctcg acttcgctgc tacccaaggt tgccgggtga cgcacaccgt ggaaacggat
     2401 gaaggcacga acccagtgga cataagcctg ttcggttcgt aagctgtaat gcaagtagcg
     2461 tatgcgctca cgcaactggt ccagaacctt gaccgaacgc agcggtggta acggcgcagt
     2521 ggcggttttc atggcttgtt atgactgttt ttttggggta cagtctatgc ctcgggcatc
     2581 caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt
     2641 tacgcagcag ggcagtcgcc ctaaaacaaa gttaaacatt atgagggaag cggtgatcgc
     2701 cgaagtatcg actcaactat cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac
     2761 gttgctggcc gtacatttgt acggctccgc agtggatggc ggcctgaagc cacacagtga
     2821 tattgatttg ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc gagctttgat
     2881 caacgacctt ttggaaactt cggcttcccc tggagagagc gagattctcc gcgctgtaga
     2941 agtcaccatt gttgtgcacg acgacatcat tccgtggcgt tatccagcta agcgcgaact
     3001 gcaatttgga gaatggcagc gcaatgacat tcttgcaggt atcttcgagc cagccacgat
     3061 cgacattgat ctggctatct tgctgacaaa agcaagagaa catagcgttg ccttggtagg
     3121 tccagcggcg gaggaactct ttgatccggt tcctgaacag gatctatttg aggcgctaaa
     3181 tgaaacctta acgctatgga actcgccgcc cgactgggct ggcgatgagc gaaatgtagt
     3241 gcttacgttg tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt
     3301 cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca tacttgaagc
     3361 tagacaggct tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga
     3421 agaatttgtc cactacgtga aaggcgagat caccaaggta gtcggcaaat aaccctcgac
     3481 tctagaggat ccccgggtac cgagctcgaa ttccgtgata ggtgggctgc ccttcctggt
     3541 tggcttggtt tcatcagcca tccgcttgcc ctcatctgtt acgccggcgg tagccggcca
     3601 gcctcgcaga gcaggattcc cgttgagcac cgccaggtgc gaataaggga cagtgaagaa
     3661 ggaacacccg ctcgcgggtg ggcctacttc acctatcctg cccggctgac gccgttggat
     3721 acaccaagga aagtctacac gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga
     3781 tggatatacc gaaaaaatcg ctataatgac cccgaagcag ggttatgcag cggaaaacgg
     3841 aattcgagct cggtacccgg ggatcctcta gagtcgagcc acccatgacc aaaatccctt
     3901 aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3961 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4021 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4081 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4141 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4201 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4261 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4321 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4381 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4441 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4501 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4561 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gtt
//
