LOCUS       sequence-125843-        6419 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway ORF clone of human IRS1 [NM_005544.2] without stop codon 
            (for C-terminal fusions).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6419)
  TITLE     Esposito lab
REFERENCE   2  (bases 1 to 6419)
  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-E110 Hs.IRS1-nostop
FEATURES             Location/Qualifiers
     source          1..6419
                     /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(3910..4009)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             4386..5177
                     /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      5270..5858
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5759..5778
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6012..6029
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(6188..6215)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6307..6393)
                     /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 gtcaactttc ttgtacaaag ttggcattat aggaaagcat tgcttatcaa
     3961 tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc atgcaagctt
     4021 agatcctcta gaccagccag gacagaaatg cctcgacttc gctgctaccc aaggttgccg
     4081 ggtgacgcac accgtggaaa cggatgaagg cacgaaccca gtggacataa gcctgttcgg
     4141 ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga accttgaccg
     4201 aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttg
     4261 gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt
     4321 tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttaa
     4381 acattatgag ggaagcggtg atcgccgaag tatcgactca actatcagag gtagttggcg
     4441 tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg
     4501 atggcggcct gaagccacac agtgatattg atttgctggt tacggtgacc gtaaggcttg
     4561 atgaaacaac gcggcgagct ttgatcaacg accttttgga aacttcggct tcccctggag
     4621 agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac atcattccgt
     4681 ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat gacattcttg
     4741 caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg acaaaagcaa
     4801 gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat ccggttcctg
     4861 aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg ccgcccgact
     4921 gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac agcgcagtaa
     4981 ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc ctgccggccc
     5041 agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa gaagatcgct
     5101 tggcctcgcg cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc gagatcacca
     5161 aggtagtcgg caaataaccc tcgagccacc catgaccaaa atcccttaac gtgagttacg
     5221 cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5281 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5341 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5401 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5461 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     5521 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5581 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5641 tgagatacct acagcgtgag cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5701 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5761 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5821 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     5881 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     5941 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     6001 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     6061 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     6121 aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa gagtttgtag
     6181 aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta
     6241 tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca aatccgctcc
     6301 cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc
     6361 ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcg
//
