LOCUS       sequence-125869-        3317 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway ORF clone of human MRAS [NM_001085049.2] without stop codon 
            (for C-terminal fusions).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3317)
  TITLE     Esposito lab
REFERENCE   2  (bases 1 to 3317)
  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-E136 Hs.MRAS-nostop
FEATURES             Location/Qualifiers
     source          1..3317
                     /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(808..907)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1284..2075
                     /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      2168..2756
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2657..2676
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2910..2927
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3086..3113)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3205..3291)
                     /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 caactttctt gtacaaagtt ggcattatag
      841 gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca
      901 ttatttgcat gcaagcttag atcctctaga ccagccagga cagaaatgcc tcgacttcgc
      961 tgctacccaa ggttgccggg tgacgcacac cgtggaaacg gatgaaggca cgaacccagt
     1021 ggacataagc ctgttcggtt cgtaagctgt aatgcaagta gcgtatgcgc tcacgcaact
     1081 ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt
     1141 gttatgactg tttttttggg gtacagtcta tgcctcgggc atccaagcag caagcgcgtt
     1201 acgccgtggg tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagggcagtc
     1261 gccctaaaac aaagttaaac attatgaggg aagcggtgat cgccgaagta tcgactcaac
     1321 tatcagaggt agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt
     1381 tgtacggctc cgcagtggat ggcggcctga agccacacag tgatattgat ttgctggtta
     1441 cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa
     1501 cttcggcttc ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc
     1561 acgacgacat cattccgtgg cgttatccag ctaagcgcga actgcaattt ggagaatggc
     1621 agcgcaatga cattcttgca ggtatcttcg agccagccac gatcgacatt gatctggcta
     1681 tcttgctgac aaaagcaaga gaacatagcg ttgccttggt aggtccagcg gcggaggaac
     1741 tctttgatcc ggttcctgaa caggatctat ttgaggcgct aaatgaaacc ttaacgctat
     1801 ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca
     1861 tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa
     1921 tggagcgcct gccggcccag tatcagcccg tcatacttga agctagacag gcttatcttg
     1981 gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gtccactacg
     2041 tgaaaggcga gatcaccaag gtagtcggca aataaccctc gagccaccca tgaccaaaat
     2101 cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     2161 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     2221 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     2281 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     2341 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     2401 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     2461 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     2521 gaacgaccta caccgaactg agatacctac agcgtgagca ttgagaaagc gccacgcttc
     2581 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     2641 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     2701 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     2761 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     2821 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     2881 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     2941 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     3001 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta
     3061 gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttagt
     3121 ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac
     3181 aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca
     3241 acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt gatgcctggc
     3301 agttccctac tctcgcg
//
