LOCUS       sequence-125971-        5726 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway ORF clone of codon optimized human RASAL3 [NM_022904.1] 
            without stop codon (for C-terminal fusions).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5726)
  TITLE     Esposito lab
REFERENCE   2  (bases 1 to 5726)
  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-E238 Hs.RASAL3-nostop
FEATURES             Location/Qualifiers
     source          1..5726
                     /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(3217..3316)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3693..4484
                     /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      4577..5165
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5066..5085
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5319..5336
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5495..5522)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5614..5700)
                     /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 gcaccatgga tcccccgtcc ccctcacgga cgtcccagac ccaacctacc gcgacctccc
      241 cccttacttc ctaccgctgg cacaccggtg gcgggggaga aaaggcagca ggggggttta
      301 gatggggccg cttcgccgga tggggccggg ccctgtcgca ccaggagccc atggtgtcca
      361 cccaacccgc acctcgctcc atcttccgcc gggtgctgtc cgcgccgcct aaggagtccc
      421 gaacttcgcg gctgagactt tcaaaggccc tctgggggcg acacaagaac ccgccacccg
      481 agccggaccc tgaacccgaa caggaagccc ccgaacttga gcctgaaccg gaactcgagc
      541 cacccacccc gcaaatccct gaggctccaa ccccgaacgt gccggtctgg gatattggcg
      601 gattcacact gctggacgga aagctggtgc tgctgggagg agaggaagag ggaccgcgca
      661 gaccccgcgt gggatccgcc agctcggagg ggtccattca cgtggccatg ggaaacttcc
      721 gggatccgga ccggatgccg ggaaagaccg agcccgaaac tgccggcccg aaccaggtcc
      781 acaacgtccg gggccttctg aagcggttga aggagaagaa gaaggcccgg ctggaaccgc
      841 gggatggccc gccgtccgcc ctgggttcgc gggagagcct tgccaccctg agcgagctcg
      901 acctgggcgc cgaaagggac gtgcggatct ggcctctgca cccgtcgctg ctgggcgaac
      961 cacactgctt ccaagtcacc tggaccggag gctcaagatg cttctcgtgc cgctcggctg
     1021 ccgaacggga ccgctggatc gaggacctga ggcgccagtt ccagcctacc caggacaacg
     1081 tggagcggga ggagacttgg ctgtctgtct gggtgcacga agccaagggc ctgccgagag
     1141 ccgcagccgg agctcccggc gtgcgcgctg aactgtggct tgacggagcc ttgctggcta
     1201 gaaccgctcc gcgggccgga cctggacagc tgttttgggc cgagcgcttc catttcgaag
     1261 ccctgccgcc tgcccggcgc ctctcgctga gactccgggg cctggggccc ggaagcgccg
     1321 tgctgggacg cgtggcgctc gcactggaag aactcgacgc ccctcgggcg ccggccgcgg
     1381 gcctggagcg gtggttcccc ctcctgggtg ccccggctgg cgcggccctg agagcgcgca
     1441 tccgcgcgcg gcgcctccgc gtgctgccat ccgaacggta caaggagctc gccgagttcc
     1501 tgaccttcca ctacgcgcgg ctgtgcggcg ccctcgagcc agcattgccg gctcaggcca
     1561 aggaggaact ggccgcagcc atggtccgag tgctgagggc cactggaagg gctcaggctc
     1621 ttgtgaccga cctgggaact gccgaactgg cgcgctgtgg aggaagggaa gccctgctgt
     1681 ttcgcgaaaa caccctggcc accaaggcca tcgacgagta catgaaactg gtggcccagg
     1741 attacttgca agagactctc ggacaggtcg tgcggcgcct ctgtgcatcc accgaggact
     1801 gcgaggtcga cccatccaag tgcccggcct cagaactgcc ggaacatcag gcccgcctga
     1861 gaaactcgtg cgaagaggtg ttcgaaacca tcatccattc ttatgactgg ttccctgcgg
     1921 agctgggaat tgtgtttagc tcatggcggg aggcctgtaa agagaggggt tccgaagtgc
     1981 tcggtcctag acttgtctgt gccagcctgt tcctccggct gctctgcccg gcgatcctcg
     2041 cgccaagcct gttcggactt gctcccgatc atcccgcccc gggtcctgct aggaccttga
     2101 ccctcattgc taaggtcatc caaaacctgg ccaatagagc ccctttcggg gaaaaggaag
     2161 cttacatggg cttcatgaac agcttcctgg aggaacacgg ccctgccatg cagtgcttcc
     2221 tggatcaagt ggccatggtg gacgtggacg ccgcgccgag cggttaccaa ggcagcgggg
     2281 atctcgccct ccaactggcc gtgttgcacg cccagctttg caccattttc gctgagctgg
     2341 accagaccac cagggacacc ctggagccct tgcccacgat tctgagagcg attgaagagg
     2401 ggcagccggt gctggtgtca gtgccaatgc gcctgccgct cccacccgcc caagtgcata
     2461 gctctctgtc cgcgggagaa aagcccggat tcctggctcc ccgcgatttg cctaagcaca
     2521 ccccgctgat cagcaaatca cagtccctgc gctccgtgag aagatcagaa agctgggcta
     2581 gacctcggcc tgacgaagaa aggccactga ggcggccacg gcccgtccag aggacccaat
     2641 ccgtgccagt gcgccgccct gcccgccggc ggcagtccgc aggaccgtgg cctcggccta
     2701 agggctcgct ctccatggga ccggccccta gagccaggcc ttggaccagg gacagcgcct
     2761 cgctcccgcg gaaaccatca gtgccgtggc agagacaaat ggaccagcca caggacagaa
     2821 accaggccct gggaactcat cggcccgtga acaagctggc agaattgcag tgtgaagtgg
     2881 ccgcgctgcg cgaagaacag aaggtcctga gccgcctggt ggagtccctg tcgacccaga
     2941 tcagggccct cactgaacag caggagcagt tgcggggcca gctccaagac ctggactccc
     3001 ggctgagggc cggttcctcc gaattcgact cggagcacaa cctgacctcc aacgaaggac
     3061 acagcctgaa gaacctcgag caccgcctga acgaaatgga gcgaactcaa gcccagctca
     3121 gggatgccgt gcagagcctg cagctgtccc ctcggactcg ggggagctgg tcacagccgc
     3181 agcccctgaa ggccccgtgc ctgaacgggg atactacttc aactttcttg tacaaagttg
     3241 gcattatagg aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa
     3301 ataaaatcat tatttgcatg caagcttaga tcctctagac cagccaggac agaaatgcct
     3361 cgacttcgct gctacccaag gttgccgggt gacgcacacc gtggaaacgg atgaaggcac
     3421 gaacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag cgtatgcgct
     3481 cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt
     3541 tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc
     3601 aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc
     3661 agggcagtcg ccctaaaaca aagttaaaca ttatgaggga agcggtgatc gccgaagtat
     3721 cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg
     3781 ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt
     3841 tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc
     3901 ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca
     3961 ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg
     4021 gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg
     4081 atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg
     4141 cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct
     4201 taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt
     4261 tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg
     4321 actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg
     4381 cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg
     4441 tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg agccacccat
     4501 gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa
     4561 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4621 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4681 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     4741 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4801 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4861 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4921 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcat tgagaaagcg
     4981 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5041 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5101 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5161 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5221 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     5281 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     5341 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     5401 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatacgc
     5461 gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt
     5521 ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg
     5581 ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga gagcgttcac
     5641 cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc gttttatttg
     5701 atgcctggca gttccctact ctcgcg
//
