LOCUS       sequence-468738-        4868 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Gateway ORF Entry clone of human SMAD4 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 4868)
  TITLE     Esposito lab plasmids 2025
REFERENCE   2  (bases 1 to 4868)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: 8061-E10
FEATURES             Location/Qualifiers
     source          1..4868
                     /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(2349..2448)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2465..2484)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2466..2484)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2489..2505)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2489..2505)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2936..3727
                     /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            3849..3958
                     /note="incP origin of transfer"
     rep_origin      4218..4806
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4707..4726
                     /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 catggacaat atgtctatta cgaatacacc
      721 aacaagtaat gatgcctgtc tgagcattgt gcatagtttg atgtgccata gacaaggtgg
      781 agagagtgaa acatttgcaa aaagagcaat tgaaagtttg gtaaagaagc tgaaggagaa
      841 aaaagatgaa ttggattctt taataacagc tataactaca aatggagctc atcctagtaa
      901 atgtgttacc atacagagaa cattggatgg gaggcttcag gtggctggtc ggaaaggatt
      961 tcctcatgtg atctatgccc gtctctggag gtggcctgat cttcacaaaa atgaactaaa
     1021 acatgttaaa tattgtcagt atgcgtttga cttaaaatgt gatagtgtct gtgtgaatcc
     1081 atatcactac gaacgagttg tatcacctgg aattgatctc tcaggattaa cactgcagag
     1141 taatgctcca tcaagtatga tggtgaagga tgaatatgtg catgactttg agggacagcc
     1201 atcgttgtcc actgaaggac attcaattca aaccatccag catccaccaa gtaatcgtgc
     1261 atcgacagag acatacagca ccccagctct gttagcccca tctgagtcta atgctaccag
     1321 cactgccaac tttcccaaca ttcctgtggc ttccacaagt cagcctgcca gtatactggg
     1381 gggcagccat agtgaaggac tgttgcagat agcatcaggg cctcagccag gacagcagca
     1441 gaatggattt actggtcagc cagctactta ccatcataac agcactacca cctggactgg
     1501 aagtaggact gcaccataca cacctaattt gcctcaccac caaaacggcc atcttcagca
     1561 ccacccgcct atgccgcccc atcccggaca ttactggcct gttcacaatg agcttgcatt
     1621 ccagcctccc atttccaatc atcctgctcc tgagtattgg tgttccattg cttactttga
     1681 aatggatgtt caggtaggag agacatttaa ggttccttca agctgcccta ttgttactgt
     1741 tgatggatac gtggaccctt ctggaggaga tcgcttttgt ttgggtcaac tctccaatgt
     1801 ccacaggaca gaagccattg agagagcaag gttgcacata ggcaaaggtg tgcagttgga
     1861 atgtaaaggt gaaggtgatg tttgggtcag gtgccttagt gaccacgcgg tctttgtaca
     1921 gagttactac ttagacagag aagctgggcg tgcacctgga gatgctgttc ataagatcta
     1981 cccaagtgca tatataaagg tctttgattt gcgtcagtgt catcgacaga tgcagcagca
     2041 ggcggctact gcacaagctg cagcagctgc ccaggcagca gccgtggcag gaaacatccc
     2101 tggcccagga tcagtaggtg gaatagctcc agctatcagt ctgtcagctg ctgctggaat
     2161 tggtgttgat gaccttcgtc gcttatgcat actcaggatg agttttgtga aaggctgggg
     2221 accggattac ccaagacaga gcatcaaaga aacaccttgc tggattgaaa ttcacttaca
     2281 ccgggccctc cagctcctag acgaagtact tcataccatg ccgattgcag acccacaacc
     2341 tttagactaa tcaactttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     2401 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     2461 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     2521 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gcctcctcta
     2581 gaccagccag gacagaaatg cctcgacttc gctgctaccc aaggttgccg ggtgacgcac
     2641 accgtggaaa cggatgaagg cacgaaccca gtggacataa gcctgttcgg ttcgtaagct
     2701 gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg
     2761 tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttg gggtacagtc
     2821 tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg
     2881 gagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttaa acattatgag
     2941 ggaagcggtg atcgccgaag tatcgactca actatcagag gtagttggcg tcatcgagcg
     3001 ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg atggcggcct
     3061 gaagccacac agtgatattg atttgctggt tacggtgacc gtaaggcttg atgaaacaac
     3121 gcggcgagct ttgatcaacg accttttgga aacttcggct tcccctggag agagcgagat
     3181 tctccgcgct gtagaagtca ccattgttgt gcacgacgac atcattccgt ggcgttatcc
     3241 agctaagcgc gaactgcaat ttggagaatg gcagcgcaat gacattcttg caggtatctt
     3301 cgagccagcc acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag
     3361 cgttgccttg gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct
     3421 atttgaggcg ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga
     3481 tgagcgaaat gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat
     3541 cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc
     3601 cgtcatactt gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg
     3661 cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg
     3721 caaataaccc tcgactctag aggatccccg ggtaccgagc tcgaattccg tgataggtgg
     3781 gctgcccttc ctggttggct tggtttcatc agccatccgc ttgccctcat ctgttacgcc
     3841 ggcggtagcc ggccagcctc gcagagcagg attcccgttg agcaccgcca ggtgcgaata
     3901 agggacagtg aagaaggaac acccgctcgc gggtgggcct acttcaccta tcctgcccgg
     3961 ctgacgccgt tggatacacc aaggaaagtc tacacgaacc ctttggcaaa atcctgtata
     4021 tcgtgcgaaa aaggatggat ataccgaaaa aatcgctata atgaccccga agcagggtta
     4081 tgcagcggaa aacggaattc gagctcggta cccggggatc ctctagagtc gagccaccca
     4141 tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa
     4201 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4261 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4321 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     4381 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4441 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4501 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4561 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4621 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4681 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4741 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4801 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     4861 cacatgtt
//
