LOCUS       sequence-468852-        4721 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway ORF Entry clone of human Tgfbr1 with stop codon (for native 
            or N-terminal fusions); T204D mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4721)
  TITLE     Esposito lab plasmids 2025
REFERENCE   2  (bases 1 to 4721)
  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: 9336-E05
FEATURES             Location/Qualifiers
     source          1..4721
                     /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(2202..2301)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2318..2337)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2319..2337)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2342..2358)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2342..2358)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2789..3580
                     /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            3702..3811
                     /note="incP origin of transfer"
     rep_origin      4071..4659
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4560..4579
                     /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 catggaggcg gcggtcgctg ctccgcgtcc
      721 ccggctgctc ctcctcgtgc tggcggcggc ggcggcggcg gcggcggcgc tgctcccggg
      781 ggcgacggcg ttacagtgtt tctgccacct ctgtacaaaa gacaatttta cttgtgtgac
      841 agatgggctc tgctttgtct ctgtcacaga gaccacagac aaagttatac acaacagcat
      901 gtgtatagct gaaattgact taattcctcg agataggccg tttgtatgtg caccctcttc
      961 aaaaactggg tctgtgacta caacatattg ctgcaatcag gaccattgca ataaaataga
     1021 acttccaact actgtaaagt catcacctgg ccttggtcct gtggaactgg cagctgtcat
     1081 tgctggacca gtgtgcttcg tctgcatctc actcatgttg atggtctata tctgccacaa
     1141 ccgcactgtc attcaccatc gagtgccaaa tgaagaggac ccttcattag atcgcccttt
     1201 tatttcagag ggtactacgt tgaaagactt aatttatgat atgacaacgt caggttctgg
     1261 ctcaggttta ccattgcttg ttcagagaac aattgcgaga gatattgtgt tacaagaaag
     1321 cattggcaaa ggtcgatttg gagaagtttg gagaggaaag tggcggggag aagaagttgc
     1381 tgttaagata ttctcctcta gagaagaacg ttcgtggttc cgtgaggcag agatttatca
     1441 aactgtaatg ttacgtcatg aaaacatcct gggatttata gcagcagaca ataaagacaa
     1501 tggtacttgg actcagctct ggttggtgtc agattatcat gagcatggat ccctttttga
     1561 ttacttaaac agatacacag ttactgtgga aggaatgata aaacttgctc tgtccacggc
     1621 gagcggtctt gcccatcttc acatggagat tgttggtacc caaggaaagc cagccattgc
     1681 tcatagagat ttgaaatcaa agaatatctt ggtaaagaag aatggaactt gctgtattgc
     1741 agacttagga ctggcagtaa gacatgattc agccacagat accattgata ttgctccaaa
     1801 ccacagagtg ggaacaaaaa ggtacatggc ccctgaagtt ctcgatgatt ccataaatat
     1861 gaaacatttt gaatccttca aacgtgctga catctatgca atgggcttag tattctggga
     1921 aattgctcga cgatgttcca ttggtggaat tcatgaagat taccaactgc cttattatga
     1981 tcttgtacct tctgacccat cagttgaaga aatgagaaaa gttgtttgtg aacagaagtt
     2041 aaggccaaat atcccaaaca gatggcagag ctgtgaagcc ttgagagtaa tggctaaaat
     2101 tatgagagaa tgttggtatg ccaatggagc agctaggctt acagcattgc ggattaagaa
     2161 aacattatcg caactcagtc aacaggaagg catcaaaatg taatcaactt tcttgtacaa
     2221 agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     2281 tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat
     2341 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     2401 acaagataaa aatatatcat catgcctcct ctagaccagc caggacagaa atgcctcgac
     2461 ttcgctgcta cccaaggttg ccgggtgacg cacaccgtgg aaacggatga aggcacgaac
     2521 ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta tgcgctcacg
     2581 caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat
     2641 ggcttgttat gactgttttt ttggggtaca gtctatgcct cgggcatcca agcagcaagc
     2701 gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcaggg
     2761 cagtcgccct aaaacaaagt taaacattat gagggaagcg gtgatcgccg aagtatcgac
     2821 tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt
     2881 acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagtgata ttgatttgct
     2941 ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca acgacctttt
     3001 ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag tcaccattgt
     3061 tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc aatttggaga
     3121 atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg acattgatct
     3181 ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc cagcggcgga
     3241 ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg aaaccttaac
     3301 gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc ttacgttgtc
     3361 ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg
     3421 ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta gacaggctta
     3481 tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag aatttgtcca
     3541 ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa ccctcgactc tagaggatcc
     3601 ccgggtaccg agctcgaatt ccgtgatagg tgggctgccc ttcctggttg gcttggtttc
     3661 atcagccatc cgcttgccct catctgttac gccggcggta gccggccagc ctcgcagagc
     3721 aggattcccg ttgagcaccg ccaggtgcga ataagggaca gtgaagaagg aacacccgct
     3781 cgcgggtggg cctacttcac ctatcctgcc cggctgacgc cgttggatac accaaggaaa
     3841 gtctacacga accctttggc aaaatcctgt atatcgtgcg aaaaaggatg gatataccga
     3901 aaaaatcgct ataatgaccc cgaagcaggg ttatgcagcg gaaaacggaa ttcgagctcg
     3961 gtacccgggg atcctctaga gtcgagccac ccatgaccaa aatcccttaa cgtgagttac
     4021 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4081 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4141 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4201 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     4261 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4321 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4381 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4441 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4501 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4561 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     4621 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     4681 ttacggttcc tggccttttg ctggcctttt gctcacatgt t
//
