LOCUS       sequence-469778-        4607 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway ORF Entry clone of human SMAD1 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-E07
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 catgaatgtg acaagtttat tttcctttac
      721 aagtccagct gtgaagagac ttcttgggtg gaaacagggc gatgaagaag aaaaatgggc
      781 agagaaagct gttgatgctt tggtgaaaaa actgaagaaa aagaaaggtg ccatggagga
      841 actggaaaag gccttgagct gcccagggca accgagtaac tgtgtcacca ttccccgctc
      901 tctggatggc aggctgcaag tctcccaccg gaagggactg cctcatgtca tttactgccg
      961 tgtgtggcgc tggcccgatc ttcagagcca ccatgaacta aaaccactgg aatgctgtga
     1021 gtttcctttt ggttccaagc agaaggaggt ctgcatcaat ccctaccact ataagagagt
     1081 agaaagccct gtacttcctc ctgtgctggt tccaagacac agcgaatata atcctcagca
     1141 cagcctctta gctcagttcc gtaacttagg acaaaatgag cctcacatgc cactcaacgc
     1201 cacttttcca gattctttcc agcaacccaa cagccacccg tttcctcact ctcccaatag
     1261 cagttaccca aactctcctg ggagcagcag cagcacctac cctcactctc ccaccagctc
     1321 agacccagga agccctttcc agatgccagc tgatacgccc ccacctgctt acctgcctcc
     1381 tgaagacccc atgacccagg atggctctca gccgatggac acaaacatga tggcgcctcc
     1441 cctgccctca gaaatcaaca gaggagatgt tcaggcggtt gcttatgagg aaccaaaaca
     1501 ctggtgctct attgtctact atgagctcaa caatcgtgtg ggtgaagcgt tccatgcctc
     1561 ctccacaagt gtgttggtgg atggtttcac tgatccttcc aacaataaga accgtttctg
     1621 ccttgggctg ctctccaatg ttaaccggaa ttccactatt gaaaacacca ggcggcatat
     1681 tggaaaagga gttcatcttt attatgttgg aggggaggtg tatgccgaat gccttagtga
     1741 cagtagcatc tttgtgcaaa gtcggaactg caactaccat catggatttc atcctactac
     1801 tgtttgcaag atccctagtg ggtgtagtct gaaaattttt aacaaccaag aatttgctca
     1861 gttattggca cagtctgtga accatggatt tgagacagtc tatgagctta caaaaatgtg
     1921 tactatacgt atgagctttg tgaagggctg gggagcagaa taccaccgcc aggatgttac
     1981 tagcaccccc tgctggattg agatacatct gcacggcccc ctccagtggc tggataaagt
     2041 tcttactcaa atgggttcac ctcataatcc tatttcatct gtatcttaat 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
//
