LOCUS       sequence-125970-        5729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway ORF clone of codon optimized human RASAL3 [NM_022904.1] 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 5729)
  TITLE     Esposito lab
REFERENCE   2  (bases 1 to 5729)
  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-E237 Hs.RASAL3
FEATURES             Location/Qualifiers
     source          1..5729
                     /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(3220..3319)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3696..4487
                     /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      4580..5168
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5069..5088
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5322..5339
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5498..5525)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5617..5703)
                     /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 atactactta atcaactttc ttgtacaaag
     3241 ttggcattat aggaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc
     3301 aaaataaaat cattatttgc atgcaagctt agatcctcta gaccagccag gacagaaatg
     3361 cctcgacttc gctgctaccc aaggttgccg ggtgacgcac accgtggaaa cggatgaagg
     3421 cacgaaccca gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc
     3481 gctcacgcaa ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg
     3541 ttttcatggc ttgttatgac tgtttttttg gggtacagtc tatgcctcgg gcatccaagc
     3601 agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc
     3661 agcagggcag tcgccctaaa acaaagttaa acattatgag ggaagcggtg atcgccgaag
     3721 tatcgactca actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc
     3781 tggccgtaca tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg
     3841 atttgctggt tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg
     3901 accttttgga aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca
     3961 ccattgttgt gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat
     4021 ttggagaatg gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca
     4081 ttgatctggc tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag
     4141 cggcggagga actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa
     4201 ccttaacgct atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta
     4261 cgttgtcccg catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg
     4321 ccgactgggc aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac
     4381 aggcttatct tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat
     4441 ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg caaataaccc tcgagccacc
     4501 catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag
     4561 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4621 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4681 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     4741 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4801 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4861 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4921 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag cattgagaaa
     4981 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5041 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5101 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5161 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5221 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     5281 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     5341 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     5401 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata
     5461 cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc
     5521 cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg
     5581 ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt
     5641 caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat
     5701 ttgatgcctg gcagttccct actctcgcg
//
