LOCUS       sequence-126088-        8048 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway ORF clone of human TSC2 [NM_001114382.1] 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 8048)
  TITLE     Esposito lab
REFERENCE   2  (bases 1 to 8048)
  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-E355 Hs.TSC2
FEATURES             Location/Qualifiers
     source          1..8048
                     /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(5539..5638)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             6015..6806
                     /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      6899..7487
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7388..7407
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7641..7658
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(7817..7844)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(7936..8022)
                     /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 gtttgtgtaa tcaactttct tgtacaaagt tggcattata ggaaagcatt
     5581 gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgca
     5641 tgcaagctta gatcctctag accagccagg acagaaatgc ctcgacttcg ctgctaccca
     5701 aggttgccgg gtgacgcaca ccgtggaaac ggatgaaggc acgaacccag tggacataag
     5761 cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa
     5821 ccttgaccga acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact
     5881 gtttttttgg ggtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg
     5941 gtcgatgttt gatgttatgg agcagcaacg atgttacgca gcagggcagt cgccctaaaa
     6001 caaagttaaa cattatgagg gaagcggtga tcgccgaagt atcgactcaa ctatcagagg
     6061 tagttggcgt catcgagcgc catctcgaac cgacgttgct ggccgtacat ttgtacggct
     6121 ccgcagtgga tggcggcctg aagccacaca gtgatattga tttgctggtt acggtgaccg
     6181 taaggcttga tgaaacaacg cggcgagctt tgatcaacga ccttttggaa acttcggctt
     6241 cccctggaga gagcgagatt ctccgcgctg tagaagtcac cattgttgtg cacgacgaca
     6301 tcattccgtg gcgttatcca gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg
     6361 acattcttgc aggtatcttc gagccagcca cgatcgacat tgatctggct atcttgctga
     6421 caaaagcaag agaacatagc gttgccttgg taggtccagc ggcggaggaa ctctttgatc
     6481 cggttcctga acaggatcta tttgaggcgc taaatgaaac cttaacgcta tggaactcgc
     6541 cgcccgactg ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc atttggtaca
     6601 gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca atggagcgcc
     6661 tgccggccca gtatcagccc gtcatacttg aagctagaca ggcttatctt ggacaagaag
     6721 aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt tgtccactac gtgaaaggcg
     6781 agatcaccaa ggtagtcggc aaataaccct cgagccaccc atgaccaaaa tcccttaacg
     6841 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6901 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6961 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     7021 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     7081 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     7141 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     7201 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     7261 acaccgaact gagataccta cagcgtgagc attgagaaag cgccacgctt cccgaaggga
     7321 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     7381 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     7441 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     7501 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     7561 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     7621 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     7681 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     7741 ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag
     7801 agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct
     7861 ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa
     7921 atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa
     7981 acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta
     8041 ctctcgcg
//
