LOCUS       sequence-469771-        4607 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway ORF Entry clone of mouse Smad5 with stop codon (for native 
            or N-terminal fusions).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4607)
  TITLE     Esposito lab plasmids 2025
REFERENCE   2  (bases 1 to 4607)
  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: 8061-E35
FEATURES             Location/Qualifiers
     source          1..4607
                     /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(2088..2187)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2204..2223)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2205..2223)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2228..2244)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2228..2244)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2675..3466
                     /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            3588..3697
                     /note="incP origin of transfer"
     rep_origin      3957..4545
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4446..4465
                     /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 catgacgtca atggccagct tgttttcttt
      721 cactagtcca gccgtgaagc gattgttggg ctggaaacaa ggtgacgagg aagagaaatg
      781 ggcagaaaag gcagtggatg ctttagtgaa aaagctgaag aagaagaagg gtgctatgga
      841 ggagctggag aaagccttga gcagcccagg acagccaagc aagtgtgtca cgatccccag
      901 gtccttggat ggacgtctgc aagtttctca caggaaaggc ttgccccatg ttatatattg
      961 ccgtgtttgg cgctggccag atttgcagag ccatcacgag ctaaaaccat tggatatttg
     1021 tgaatttcct tttggatcta agcaaaagga agtttgtatc aatccatacc actataagag
     1081 agtggagagt ccagtcttac ctccagtatt agtgcctcgt cacaatgaat tcaatccaca
     1141 acacagcctt ctggttcagt tcaggaacct gagccacaat gaaccgcaca tgccacaaaa
     1201 cgccacgttt cccgattctt tccaccaacc caacaacgct cctttcccct tatctcctaa
     1261 cagcccctat cctccttccc ctgctagcag cacatatccc aactccccag caagctctgg
     1321 acctggaagt ccatttcaac tcccagctga cacccctccc cctgcctata tgccacctga
     1381 tgatcagatg gccccagata attcccagcc tatggataca agcagtaaca tgattcctca
     1441 gaccatgccc agcatatcca gcagagatgt tcagcctgtc gcctatgagg agcccaaaca
     1501 ctggtgttcg attgtctact atgaattaaa caatcgtgtt ggggaagctt ttcatgcatc
     1561 ttctactagt gtgttagtag atggatttac agatccttca aataacaaaa gtagattctg
     1621 cctgggattg ttgtcaaatg ttaatcgtaa ttcaactatt gaaaacacta ggcggcatat
     1681 tggaaaaggt gttcatctat actacgttgg tggggaggtg tacgctgagt gtcttagtga
     1741 cagcagcatc tttgttcaga gtaggaactg caactttcac catggcttcc atcccaccac
     1801 cgtctgtaag atccccagca gctgcagcct caagattttt aacaatcagg agtttgctca
     1861 gcttctggct cagtcagtca accatggatt cgaggctgtg tatgagctca ccaagatgtg
     1921 taccattcga atgagctttg tcaagggctg gggagcagag taccaccgac aggacgtcac
     1981 cagtactccc tgctggattg agattcacct ccacgggcct ctgcagtggc tggataaagt
     2041 ccttactcag atgggctctc cgctgaaccc catttcttct gtttcataat caactttctt
     2101 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     2161 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     2221 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     2281 cattgcacaa gataaaaata tatcatcatg cctcctctag accagccagg acagaaatgc
     2341 ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac ggatgaaggc
     2401 acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg
     2461 ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg cagtggcggt
     2521 tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg catccaagca
     2581 gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg atgttacgca
     2641 gcagggcagt cgccctaaaa caaagttaaa cattatgagg gaagcggtga tcgccgaagt
     2701 atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac cgacgttgct
     2761 ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca gtgatattga
     2821 tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt tgatcaacga
     2881 ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg tagaagtcac
     2941 cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg aactgcaatt
     3001 tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca cgatcgacat
     3061 tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg taggtccagc
     3121 ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc taaatgaaac
     3181 cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg tagtgcttac
     3241 gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc
     3301 cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg aagctagaca
     3361 ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt
     3421 tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct cgactctaga
     3481 ggatccccgg gtaccgagct cgaattccgt gataggtggg ctgcccttcc tggttggctt
     3541 ggtttcatca gccatccgct tgccctcatc tgttacgccg gcggtagccg gccagcctcg
     3601 cagagcagga ttcccgttga gcaccgccag gtgcgaataa gggacagtga agaaggaaca
     3661 cccgctcgcg ggtgggccta cttcacctat cctgcccggc tgacgccgtt ggatacacca
     3721 aggaaagtct acacgaaccc tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata
     3781 taccgaaaaa atcgctataa tgaccccgaa gcagggttat gcagcggaaa acggaattcg
     3841 agctcggtac ccggggatcc tctagagtcg agccacccat gaccaaaatc ccttaacgtg
     3901 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3961 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4021 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4081 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     4141 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4201 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4261 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4321 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4381 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4441 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4501 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4561 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
