LOCUS       sequence-126089-        8045 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway ORF clone of human TSC2 [NM_001114382.1] without stop codon 
            (for C-terminal fusions).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8045)
  TITLE     Esposito lab
REFERENCE   2  (bases 1 to 8045)
  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: R777-E356 Hs.TSC2-nostop
FEATURES             Location/Qualifiers
     source          1..8045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..45
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     38..54
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    84..183
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(5536..5635)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             6012..6803
                     /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"
     rep_origin      6896..7484
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7385..7404
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7638..7655
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(7814..7841)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(7933..8019)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaaa
       61 attcgagctc ggtccgagga tccgaaataa tgattttatt ttgactgata gtgacctgtt
      121 cgttgcaaca aattgataag caatgctttt ttataatgcc aactttgtac aaaaaagttg
      181 gcaccatggc caaaccaaca agcaaagatt caggcttgaa ggagaagttt aagattctgt
      241 tgggactggg aacaccgagg ccaaatccca ggtctgcaga gggtaaacag acggagttta
      301 tcatcaccgc ggaaatactg agagaactga gcatggaatg tggcctcaac aatcgcatcc
      361 ggatgatagg gcagatttgt gaagtcgcaa aaaccaagaa atttgaagag cacgcagtgg
      421 aagcactctg gaaggcggtc gcggatctgt tgcagccgga gcggccgctg gaggcccggc
      481 acgcggtgct ggctctgctg aaggccatcg tgcaggggca gggcgagcgt ttgggggtcc
      541 tcagagccct cttctttaag gtcatcaagg attacccttc caacgaagac cttcacgaaa
      601 ggctggaggt tttcaaggcc ctcacagaca atgggagaca catcacctac ttggaggaag
      661 agctggctga ctttgtcctg cagtggatgg atgttggctt gtcctcggaa ttccttctgg
      721 tgctggtgaa cttggtcaaa ttcaatagct gttacctcga cgagtacatc gcaaggatgg
      781 ttcagatgat ctgtctgctg tgcgtccgga ccgcgtcctc tgtggacata gaggtctccc
      841 tgcaggtgct ggacgccgtg gtctgctaca actgcctgcc ggctgagagc ctcccgctgt
      901 tcatcgttac cctctgtcgc accatcaacg tcaaggagct ctgcgagcct tgctggaagc
      961 tgatgcggaa cctccttggc acccacctgg gccacagcgc catctacaac atgtgccacc
     1021 tcatggagga cagagcctac atggaggacg cgcccctgct gagaggagcc gtgttttttg
     1081 tgggcatggc tctctgggga gcccaccggc tctattctct caggaactcg ccgacatctg
     1141 tgttgccatc attttaccag gccatggcat gtccgaacga ggtggtgtcc tatgagatcg
     1201 tcctgtccat caccaggctc atcaagaagt ataggaagga gctccaggtg gtggcgtggg
     1261 acattctgct gaacatcatc gaacggctcc ttcagcagct ccagaccttg gacagcccgg
     1321 agctcaggac catcgtccat gacctgttga ccacggtgga ggagctgtgt gaccagaacg
     1381 agttccacgg gtctcaggag agatactttg aactggtgga gagatgtgcg gaccagaggc
     1441 ctgagtcctc cctcctgaac ctgatctcct atagagcgca gtccatccac ccggccaagg
     1501 acggctggat tcagaacctg caggcgctga tggagagatt cttcaggagc gagtcccgag
     1561 gcgccgtgcg catcaaggtg ctggacgtgc tgtcctttgt gctgctcatc aacaggcagt
     1621 tctatgagga ggagctgatt aactcagtgg tcatctcgca gctctcccac atccccgagg
     1681 ataaagacca ccaggtccga aagctggcca cccagttgct ggtggacctg gcagagggct
     1741 gccacacaca ccacttcaac agcctgctgg acatcatcga gaaggtgatg gcccgctccc
     1801 tctccccacc cccggagctg gaagaaaggg atgtggccgc atactcggcc tccttggagg
     1861 atgtgaagac agccgtcctg gggcttctgg tcatccttca gaccaagctg tacaccctgc
     1921 ctgcaagcca cgccacgcgt gtgtatgaga tgctggtcag ccacattcag ctccactaca
     1981 agcacagcta caccctgcca atcgcgagca gcatccggct gcaggccttt gacttcctgt
     2041 tgctgctgcg ggccgactca ctgcaccgcc tgggcctgcc caacaaggat ggagtcgtgc
     2101 ggttcagccc ctactgcgtc tgcgactaca tggagccaga gagaggctct gagaagaaga
     2161 ccagcggccc cctttctcct cccacagggc ctcctggccc ggcgcctgca ggccccgccg
     2221 tgcggctggg gtccgtgccc tactccctgc tcttccgcgt cctgctgcag tgcttgaagc
     2281 aggagtctga ctggaaggtg ctgaagctgg ttctgggcag gctgcctgag tccctgcgct
     2341 ataaagtgct catctttact tccccttgca gtgtggacca gctgtgctct gctctctgct
     2401 ccatgctttc aggcccaaag acactggagc ggctccgagg cgccccagaa ggcttctcca
     2461 gaactgactt gcacctggcc gtggttccag tgctgacagc attaatctct taccataact
     2521 acctggacaa aaccaaacag cgcgagatgg tctactgcct ggagcagggc ctcatccacc
     2581 gctgtgccag ccagtgcgtc gtggccttgt ccatctgcag cgtggagatg cctgacatca
     2641 tcatcaaggc gctgcctgtt ctggtggtga agctcacgca catctcagcc acagccagca
     2701 tggccgtccc actgctggag ttcctgtcca ctctggccag gctgccgcac ctctacagga
     2761 actttgccgc ggagcagtat gccagtgtgt tcgccatctc cctgccgtac accaacccct
     2821 ccaagtttaa tcagtacatc gtgtgtctgg cccatcacgt catagccatg tggttcatca
     2881 ggtgccgcct gcccttccgg aaggattttg tccctttcat cactaagggc ctgcggtcca
     2941 atgtcctctt gtcttttgat gacacccccg agaaggacag cttcagggcc cggagtacta
     3001 gtctcaacga gagacccaag agtctgagga tagccagacc ccccaaacaa ggcttgaata
     3061 actctccacc cgtgaaagaa ttcaaggaga gctctgcagc cgaggccttc cggtgccgca
     3121 gcatcagtgt gtctgaacat gtggtccgca gcaggataca gacgtccctc accagtgcca
     3181 gcttggggtc tgcagatgag aactccgtgg cccaggctga cgatagcctg aaaaacctcc
     3241 acctggagct cacggaaacc tgtctggaca tgatggctcg atacgtcttc tccaacttca
     3301 cggctgtccc gaagaggtct cctgtgggcg agttcctcct agcgggtggc aggaccaaaa
     3361 cctggctggt tgggaacaag cttgtcactg tgacgacaag cgtgggaacc gggacccggt
     3421 cgttactagg cctggactcg ggggagctgc agtccggccc ggagtcgagc tccagccccg
     3481 gggtgcatgt gagacagacc aaggaggcgc cggccaagct ggagtcccag gctgggcagc
     3541 aggtgtcccg tggggcccgg gatcgggtcc gttccatgtc ggggggccat ggtcttcgag
     3601 ttggcgccct ggacgtgccg gcctcccagt tcctgggcag tgccacttct ccaggaccac
     3661 ggactgcacc agccgcgaaa cctgagaagg cctcagctgg cacccgggtt cctgtgcagg
     3721 agaagacgaa cctggcggcc tatgtgcccc tgctgaccca gggctgggcg gagatcctgg
     3781 tccggaggcc cacagggaac accagctggc tgatgagcct ggagaacccg ctcagccctt
     3841 tctcctcgga catcaacaac atgcccctgc aggagctgtc taacgccctc atggcggctg
     3901 agcgcttcaa ggagcaccgg gacacagccc tgtacaagtc actgtcggtg ccggcagcca
     3961 gcacggccaa accccctcct ctgcctcgct ccaacacaga ctccgccgtg gtcatggagg
     4021 agggaagtcc gggcgaggtt cctgtgctgg tggagccccc agggttggag gacgttgagg
     4081 cagcgctagg catggacagg cgcacggatg cctacagcag gtcgtcctca gtctccagcc
     4141 aggaggagaa gtcgctccac gcggaggagc tggttggcag gggcatcccc atcgagcgag
     4201 tcgtctcctc ggagggtggc cggccctctg tggacctctc cttccagccc tcgcagcccc
     4261 tgagcaagtc cagctcctct cccgagctgc agactctgca ggacatcctc ggggaccctg
     4321 gggacaaggc cgacgtgggc cggctgagcc ctgaggttaa ggcccggtca cagtcaggga
     4381 ccctggacgg ggaaagtgct gcctggtcgg cctcgggcga agacagtcgg ggccagcccg
     4441 agggtccctt gccttccagc tccccccgct cgcccagtgg cctccggccc cgaggttaca
     4501 ccatctccga ctcggcccca tcacgcaggg gcaagagagt agagagggac gccttaaaga
     4561 gcagagccac agcctccaat gcagagaaag tgccaggcat caaccccagt ttcgtgttcc
     4621 tgcagctcta ccattccccc ttctttggcg acgagtcaaa caagccaatc ctgctgccca
     4681 atgagtcaca gtcctttgag cggtcggtgc agctcctcga ccagatccca tcatacgaca
     4741 cccacaagat cgccgtcctg tatgttggag aaggccagag caacagcgag ctcgccatcc
     4801 tgtccaatga gcatggctcc tacaggtaca cggagttcct gacgggcctg ggccggctca
     4861 tcgagctgaa ggactgccag ccggacaagg tgtacctggg aggcctggac gtgtgtggtg
     4921 aggacggcca gttcacctac tgctggcacg atgacatcat gcaagccgtc ttccacatcg
     4981 ccaccctgat gcccaccaag gacgtggaca agcaccgctg cgacaagaag cgccacctgg
     5041 gcaacgactt tgtgtccatt gtctacaatg actccggtga ggacttcaag cttggcacca
     5101 tcaagggcca gttcaacttt gtccacgtga tcgtcacccc gctggactac gagtgcaacc
     5161 tggtgtccct gcagtgcagg aaagacatgg agggccttgt ggacaccagc gtggccaaga
     5221 tcgtgtctga ccgcaacctg cccttcgtgg cccgccagat ggccctgcac gcaaatatgg
     5281 cctcacaggt gcatcatagc cgctccaacc ccaccgatat ctacccctcc aagtggattg
     5341 cccggctccg ccacatcaag cggctccgcc agcggatctg cgaggaagcc gcctactcca
     5401 accccagcct acctctggtg caccctccgt cccatagcaa agcccctgca cagactccag
     5461 ccgagcccac acctggctat gaggtgggcc agcggaagcg cctcatctcc tcggtggagg
     5521 acttcaccga gtttgtgtca actttcttgt acaaagttgg cattatagga aagcattgct
     5581 tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgcatgc
     5641 aagcttagat cctctagacc agccaggaca gaaatgcctc gacttcgctg ctacccaagg
     5701 ttgccgggtg acgcacaccg tggaaacgga tgaaggcacg aacccagtgg acataagcct
     5761 gttcggttcg taagctgtaa tgcaagtagc gtatgcgctc acgcaactgg tccagaacct
     5821 tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt
     5881 tttttggggt acagtctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc
     5941 gatgtttgat gttatggagc agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa
     6001 agttaaacat tatgagggaa gcggtgatcg ccgaagtatc gactcaacta tcagaggtag
     6061 ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc cgtacatttg tacggctccg
     6121 cagtggatgg cggcctgaag ccacacagtg atattgattt gctggttacg gtgaccgtaa
     6181 ggcttgatga aacaacgcgg cgagctttga tcaacgacct tttggaaact tcggcttccc
     6241 ctggagagag cgagattctc cgcgctgtag aagtcaccat tgttgtgcac gacgacatca
     6301 ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca
     6361 ttcttgcagg tatcttcgag ccagccacga tcgacattga tctggctatc ttgctgacaa
     6421 aagcaagaga acatagcgtt gccttggtag gtccagcggc ggaggaactc tttgatccgg
     6481 ttcctgaaca ggatctattt gaggcgctaa atgaaacctt aacgctatgg aactcgccgc
     6541 ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg
     6601 cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc
     6661 cggcccagta tcagcccgtc atacttgaag ctagacaggc ttatcttgga caagaagaag
     6721 atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg aaaggcgaga
     6781 tcaccaaggt agtcggcaaa taaccctcga gccacccatg accaaaatcc cttaacgtga
     6841 gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6901 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6961 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     7021 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     7081 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     7141 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     7201 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     7261 ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa
     7321 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     7381 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     7441 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     7501 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     7561 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     7621 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     7681 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     7741 actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc caggaagagt
     7801 ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt gatgcctggc
     7861 agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc
     7921 cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg
     7981 aaaggcccag tcttccgact gagcctttcg ttttatttga tgcctggcag ttccctactc
     8041 tcgcg
//
