LOCUS       sequence-125842-        6422 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway ORF clone of human IRS1 [NM_005544.2] 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 6422)
  TITLE     Esposito lab
REFERENCE   2  (bases 1 to 6422)
  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-E109 Hs.IRS1
FEATURES             Location/Qualifiers
     source          1..6422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..45
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     37..54
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     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"
     regulatory      3270..3279
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3913..4012)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             4389..5180
                     /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      5273..5861
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5762..5781
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6015..6032
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(6191..6218)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6310..6396)
                     /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 gagccctccg gagagcgatg gcttctcgga cgtgcgcaag gtgggctacc
      241 tgcgcaaacc caagagcatg cacaaacgct tcttcgtact gcgcgcggcc agcgaggctg
      301 ggggcccggc gcgcctcgag tactacgaga acgagaagaa gtggcggcac aagtcgagcg
      361 cccccaaacg ctcgatcccc cttgagagct gcttcaacat caacaagcgg gctgactcca
      421 agaacaagca cctggtggct ctctacaccc gggacgagca ctttgccatc gcggcggaca
      481 gcgaggccga gcaagacagc tggtaccagg ctctcctaca gctgcacaac cgtgctaagg
      541 gccaccacga cggagctgcg gccctcgggg cgggaggtgg tgggggcagc tgcagcggca
      601 gctccggcct tggtgaggct ggggaggact tgagctacgg tgacgtgccc ccaggacccg
      661 cattcaaaga ggtctggcaa gtgatcctga agcccaaggg cctgggtcag acaaagaacc
      721 tgattggtat ctaccgcctt tgcctgacca gcaagaccat cagcttcgtg aagctgaact
      781 cggaggcagc ggccgtggtg ctgcagctga tgaacatcag gcgctgtggc cactcggaaa
      841 acttcttctt catcgaggtg ggccgttctg ccgtgacggg gcccggggag ttctggatgc
      901 aggtggatga ctctgtggtg gcccagaaca tgcacgagac catcctggag gccatgcggg
      961 ccatgagtga tgagttccgc cctcgcagca agagccagtc ctcgtccaac tgctctaacc
     1021 ccatcagcgt ccccctgcgc cggcaccatc tcaacaatcc cccgcccagc caggtggggc
     1081 tgacccgccg atcacgcact gagagcatca ccgccacctc cccggccagc atggtgggcg
     1141 ggaagccagg ctccttccgt gtccgcgcct ccagtgacgg cgaaggcacc atgtcccgcc
     1201 cagcctcggt ggacggcagc cctgtgagtc ccagcaccaa cagaacccac gcccaccggc
     1261 atcggggcag cgcccggctg caccccccgc tcaaccacag ccgctccatc cccatgccgg
     1321 cttcccgctg ctcgccttcg gccaccagcc cggtcagtct gtcgtccagt agcaccagtg
     1381 gccatggctc cacctcggat tgtctcttcc cacggcgatc tagtgcttcg gtgtctggtt
     1441 cccccagcga tggcggtttc atctcctcgg atgagtatgg ctccagtccc tgcgatttcc
     1501 ggagttcctt ccgcagtgtc actccggatt ccctgggcca caccccacca gcccgcggtg
     1561 aggaggagct aagcaactat atctgcatgg gtggcaaggg gccctccacc ctgaccgccc
     1621 ccaacggtca ctacattttg tctcggggtg gcaatggcca ccgctgcacc ccaggaacag
     1681 gcttgggcac gagtccagcc ttggctgggg atgaagcagc cagtgctgca gatctggata
     1741 atcggttccg aaagagaact cactcggcag gcacatcccc taccattacc caccagaaga
     1801 ccccgtccca gtcctcagtg gcttccattg aggagtacac agagatgatg cctgcctacc
     1861 caccaggagg tggcagtgga ggccgactgc cgggacacag gcactccgcc ttcgtgccca
     1921 cccgctccta cccagaggag ggtctggaaa tgcacccctt ggagcgtcgg ggggggcacc
     1981 accgcccaga cagctccacc ctccacacgg atgatggcta catgcccatg tccccagggg
     2041 tggccccagt gcccagtggc cgaaagggca gtggagacta tatgcccatg agccccaaga
     2101 gcgtatctgc cccacagcag atcatcaatc ccatcagacg ccatccccag agagtggacc
     2161 ccaatggcta catgatgatg tcccccagcg gtggctgctc tcctgacatt ggaggtggcc
     2221 ccagcagcag cagcagcagc agcaacgccg tcccttccgg gaccagctat ggaaagctgt
     2281 ggacaaacgg ggtagggggc caccactctc atgtcttgcc tcaccccaaa cccccagtgg
     2341 agagcagcgg tggtaagctc ttaccttgca caggtgacta catgaacatg tcaccagtgg
     2401 gggactccaa caccagcagc ccctccgact gctactacgg ccctgaggac ccccagcaca
     2461 agccagtcct ctcctactac tcattgccaa gatcctttaa gcacacccag cgccccgggg
     2521 agccggagga gggtgcccgg catcagcacc tccgcctttc cactagctct ggtcgccttc
     2581 tctatgctgc aacagcagat gattcttcct cttccaccag cagcgacagc ctgggtgggg
     2641 gatactgcgg ggctaggctg gagcccagcc ttccacatcc ccaccatcag gttctgcagc
     2701 cccatctgcc tcgaaaggtg gacacagctg ctcagaccaa tagccgcctg gcccggccca
     2761 cgaggctgtc cctgggggat cccaaggcca gcaccttacc tcgggcccga gagcagcagc
     2821 agcagcagca gcccttgctg caccctccag agcccaagag cccgggggaa tatgtcaata
     2881 ttgaatttgg gagtgatcag tctggctact tgtctggccc ggtggctttc cacagctcac
     2941 cttctgtcag gtgtccatcc cagctccagc cagctcccag agaggaagag actggcactg
     3001 aggagtacat gaagatggac ctggggccgg gccggagggc agcctggcag gagagcactg
     3061 gggtcgagat gggcagactg ggccctgcac ctcccggggc tgctagcatt tgcaggccta
     3121 cccgggcagt gcccagcagc cggggtgact acatgaccat gcagatgagt tgtccccgtc
     3181 agagctacgt ggacacctcg ccagctgccc ctgtaagcta tgctgacatg cgaacaggca
     3241 ttgctgcaga ggaggtgagc ctgcccaggg ccaccatggc tgctgcctcc tcatcctcag
     3301 cagcctctgc ttccccgact gggcctcaag gggcagcaga gctggctgcc cactcgtccc
     3361 tgctgggggg cccacaagga cctgggggca tgagcgcctt cacccgggtg aacctcagtc
     3421 ctaaccgcaa ccagagtgcc aaagtgatcc gtgcagaccc acaagggtgc cggcggaggc
     3481 atagctccga gactttctcc tcaacaccca gtgccacccg ggtgggcaac acagtgccct
     3541 ttggagcggg ggcagcagta gggggcggtg gcggtagcag cagcagcagc gaggatgtga
     3601 aacgccacag ctctgcttcc tttgagaatg tgtggctgag gcctggggag cttgggggag
     3661 cccccaagga gccagccaaa ctgtgtgggg ctgctggggg tttggagaat ggtcttaact
     3721 acatagacct ggatttggtc aaggacttca aacagtgccc tcaggagtgc acccctgaac
     3781 cgcagcctcc cccaccccca ccccctcatc aacccctggg cagcggtgag agcagctcca
     3841 cccgccgctc aagtgaggat ttaagcgcct atgccagcat cagtttccag aagcagccag
     3901 aggaccgtca gtaatcaact ttcttgtaca aagttggcat tataggaaag cattgcttat
     3961 caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt tgcatgcaag
     4021 cttagatcct ctagaccagc caggacagaa atgcctcgac ttcgctgcta cccaaggttg
     4081 ccgggtgacg cacaccgtgg aaacggatga aggcacgaac ccagtggaca taagcctgtt
     4141 cggttcgtaa gctgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga
     4201 ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt
     4261 ttggggtaca gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat
     4321 gtttgatgtt atggagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt
     4381 taaacattat gagggaagcg gtgatcgccg aagtatcgac tcaactatca gaggtagttg
     4441 gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt acatttgtac ggctccgcag
     4501 tggatggcgg cctgaagcca cacagtgata ttgatttgct ggttacggtg accgtaaggc
     4561 ttgatgaaac aacgcggcga gctttgatca acgacctttt ggaaacttcg gcttcccctg
     4621 gagagagcga gattctccgc gctgtagaag tcaccattgt tgtgcacgac gacatcattc
     4681 cgtggcgtta tccagctaag cgcgaactgc aatttggaga atggcagcgc aatgacattc
     4741 ttgcaggtat cttcgagcca gccacgatcg acattgatct ggctatcttg ctgacaaaag
     4801 caagagaaca tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt gatccggttc
     4861 ctgaacagga tctatttgag gcgctaaatg aaaccttaac gctatggaac tcgccgcccg
     4921 actgggctgg cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag
     4981 taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg
     5041 cccagtatca gcccgtcata cttgaagcta gacaggctta tcttggacaa gaagaagatc
     5101 gcttggcctc gcgcgcagat cagttggaag aatttgtcca ctacgtgaaa ggcgagatca
     5161 ccaaggtagt cggcaaataa ccctcgagcc acccatgacc aaaatccctt aacgtgagtt
     5221 acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5281 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5341 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5401 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5461 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5521 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5581 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5641 aactgagata cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg
     5701 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5761 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5821 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5881 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     5941 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     6001 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     6061 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     6121 ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac cgctagccag gaagagtttg
     6181 tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttagtttgat gcctggcagt
     6241 ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc
     6301 tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa
     6361 ggcccagtct tccgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg
     6421 cg
//
