LOCUS       sequence-125868-        3320 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Gateway ORF clone of human MRAS [NM_001085049.2] 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 3320)
  TITLE     Esposito lab
REFERENCE   2  (bases 1 to 3320)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: R777-E135 Hs.MRAS
FEATURES             Location/Qualifiers
     source          1..3320
                     /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"
     protein_bind    complement(811..910)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1287..2078
                     /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      2171..2759
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2660..2679
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2913..2930
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3089..3116)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3208..3294)
                     /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 aaccagcgcc gtccccagtg acaacctccc cacatacaag ctggtggtgg
      241 tgggggatgg gggtgtgggc aaaagtgccc tcaccatcca gtttttccag aagatctttg
      301 tgcctgacta tgaccccacc attgaagact cctacctgaa acatacggag attgacaatc
      361 aatgggccat cttggacgtt ctggacacag ctgggcagga ggaattcagc gccatgcggg
      421 agcaatacat gcgcacgggg gatggcttcc tcatcgtcta ctccgtcact gacaaggcca
      481 gctttgagca cgtggaccgc ttccaccagc ttatcctgcg cgtcaaagac agggagtcat
      541 tcccgatgat cctcgtggcc aacaaggtcg atttgatgca cttgaggaag atcaccaggg
      601 agcaaggaaa agaaatggcg accaaacaca atattccgta catagaaacc agtgccaagg
      661 acccacctct caatgtcgac aaagccttcc atgacctcgt tagagtaatt aggcaacaga
      721 ttccggaaaa aagccagaag aagaagaaga aaaccaaatg gcggggagac cgggccacag
      781 gcacccacaa actgcaatgt gtgatcttgt aatcaacttt cttgtacaaa gttggcatta
      841 taggaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa
      901 tcattatttg catgcaagct tagatcctct agaccagcca ggacagaaat gcctcgactt
      961 cgctgctacc caaggttgcc gggtgacgca caccgtggaa acggatgaag gcacgaaccc
     1021 agtggacata agcctgttcg gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca
     1081 actggtccag aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg
     1141 cttgttatga ctgttttttt ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc
     1201 gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagggca
     1261 gtcgccctaa aacaaagtta aacattatga gggaagcggt gatcgccgaa gtatcgactc
     1321 aactatcaga ggtagttggc gtcatcgagc gccatctcga accgacgttg ctggccgtac
     1381 atttgtacgg ctccgcagtg gatggcggcc tgaagccaca cagtgatatt gatttgctgg
     1441 ttacggtgac cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg
     1501 aaacttcggc ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc accattgttg
     1561 tgcacgacga catcattccg tggcgttatc cagctaagcg cgaactgcaa tttggagaat
     1621 ggcagcgcaa tgacattctt gcaggtatct tcgagccagc cacgatcgac attgatctgg
     1681 ctatcttgct gacaaaagca agagaacata gcgttgcctt ggtaggtcca gcggcggagg
     1741 aactctttga tccggttcct gaacaggatc tatttgaggc gctaaatgaa accttaacgc
     1801 tatggaactc gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc
     1861 gcatttggta cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg
     1921 caatggagcg cctgccggcc cagtatcagc ccgtcatact tgaagctaga caggcttatc
     1981 ttggacaaga agaagatcgc ttggcctcgc gcgcagatca gttggaagaa tttgtccact
     2041 acgtgaaagg cgagatcacc aaggtagtcg gcaaataacc ctcgagccac ccatgaccaa
     2101 aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca
     2161 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     2221 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     2281 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     2341 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     2401 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     2461 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     2521 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gcattgagaa agcgccacgc
     2581 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     2641 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     2701 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     2761 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     2821 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     2881 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     2941 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     3001 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg
     3061 ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt
     3121 agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt
     3181 cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa
     3241 acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct
     3301 ggcagttccc tactctcgcg
//
