LOCUS       sequence-125968-        6536 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway ORF clone of human RASAL2 [NM_170692.2] 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 6536)
  TITLE     Esposito lab
REFERENCE   2  (bases 1 to 6536)
  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-E235 Hs.RASAL2
FEATURES             Location/Qualifiers
     source          1..6536
                     /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(4027..4126)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             4503..5294
                     /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      5387..5975
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5876..5895
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6129..6146
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(6305..6332)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6424..6510)
                     /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 gctctctccg tcgtccggag gagccgcgga ggcgctgtcc tggccggaga
      241 tgttcccggc gctggagtcc gactcgccgc tgcccccgga ggacctggac gcggttgtcc
      301 cagtcagtgg agccgtcgcc ggtggcatgt tggatcggat ccttctggag tccgtgtgcc
      361 agcaacagag ctgggtccgg gtgtacgatg tgaaaggacc acccacccac cgtctgtctt
      421 gtggtcagtc accctacacc gagacaacaa cgtgggagcg gaagtattgc atcctcacag
      481 acagccagtt ggtattgctc aacaaggaga aggagatacc tgtggaaggg ggacaggagc
      541 agcagacaga ttccaccaaa gggcgatgcc tgaggagaac tgtcagtgtc ccttccgagg
      601 gtcagtttcc cgagtaccca ccagagggcg ccactaaact ggaggtacca gcagaaaggt
      661 cccctcgtag acggagtatc tcagggacca gtacatcaga gaaacccaac tccatggaca
      721 ctgcaaatac ctcacccttc aaagtaccag gattcttcag caagcgcctg aaaggctcca
      781 tcaagaggac caaaagccag tcaaagcttg acagaaacac gagctttcgg cttccatccc
      841 ttcgcagtac agatgacagg tctcgtgggc tgcctaaact aaaagagtca cgttcccatg
      901 aatccttgct gagcccatgc agcacagtgg aatgtctgga tcttggtaga ggggaacctg
      961 tatcagtgaa accacttcat agtagcatcc ttggacaaga cttctgcttt gaggttacct
     1021 acttaagtgg aagtaaatgc ttcagctgta attctgcttc tgagagagac aagtggatgg
     1081 aaaaccttcg caggacagtt caacctaata aggacaattg caggcgagct gaaaatgttc
     1141 ttcgtttatg gatcattgaa gccaaggacc ttgcccctaa aaagaaatat ttctgcgaac
     1201 tgtgccttga tgataccctc tttgctcgta caaccagcaa gaccaaagca gacaatattt
     1261 tctggggcga acattttgaa ttcttcagcc ttccacctct tcatagtatc acagttcaca
     1321 tttacaagga tgtggaaaaa aagaaaaaaa aggacaagaa taattatgta gggctagtca
     1381 acatccccac tgccagtgtg actggtcgcc aatttgtaga aaagtggtat ccagtgagta
     1441 cacctacacc caacaaagga aagacaggag gaccttctat tcggattaaa tcacgtttcc
     1501 aaactatcac cattctgcct atggagcaat acaaagaatt tgcagaattt gtcaccagca
     1561 actacaccat gctgtgttct gtccttgagc cagtaattag tgtgagaaat aaagaggagt
     1621 tggcttgtgc cttagtgcac attcttcaaa gtactggcag agccaaggat tttctgactg
     1681 acttggtgat gtctgaggtg gatcgttgtg gagagcatga tgtcttgatc ttcagagaga
     1741 acactattgc caccaaatcc attgaggaat acctcaagtt ggtgggacaa cagtatcttc
     1801 atgacgcact gggggagttt atcaaagctt tgtatgagtc cgatgagaac tgtgaagtgg
     1861 atcccagcaa atgttcatct agtgaactga tagaccatca gagcaacctg aaaatgtgct
     1921 gtgagctggc tttctgcaag atcatcaact cttactgtgt tttccctcgt gagttgaaag
     1981 aagtgtttgc atcatggaag cagcagtgcc tgaaccgtgg caagcaagac atcagcgaga
     2041 ggctcatcag tgcctcatta tttctccgtt ttctgtgtcc agccattatg tctcccagtc
     2101 ttttcaacct tatgcaggag tatcctgatg accgcacatc tcggactcta actcttattg
     2161 ccaaggtcat tcagaacctg gccaactttg ccaagtttgg taacaaagag gaatacatgg
     2221 cattcatgaa tgatttttta gaacatgaat ggggtggaat gaagcgcttt cttttggaga
     2281 tctctaatcc agacaccatc tcaaacaccc caggctttga tggttacatt gatctgggcc
     2341 gagagctttc agttttgcat tccttactgt gggaagtagt ttcccaactt gataaggcga
     2401 ccgtggcaaa attggggcct ctccctcgtg ttcttgctga tattaccaag tcattgacta
     2461 atcctacgcc aatacaacag caactgagac gcttcactga acataactcc agtccaaatg
     2521 tcagtggaag cctctcctct gggctgcaga aaatatttga agaccccact gacagtgatt
     2581 tgcataaact aaaatctcca agccaggaca acacagacag ctacttcaga gggaaaacat
     2641 tattgctggt tcagcaagcc tcctctcaga gcatgactta ttctgaaaag gatgaaaggg
     2701 aaagtagcct tcctaatggt cggagcgtct ccctcatgga cctccaggac actcatgctg
     2761 ctcaagtgga gcatgcatct gtcatgcttg atgtgcctat acgcttgacc ggaagccagc
     2821 tttccataac ccaggtggcc agcatcaaac agctgcggga aacccagagc actccccaaa
     2881 gtgcacccca agtgagaagg cccctgcacc cagccttgaa ccagccaggt ggccttcagc
     2941 ccttgtcgtt ccagaaccct gtctatcacc tcaataaccc aattccagca atgccaaagg
     3001 cctctataga ttccagtttg gagaacctaa gcactgccag ttccagaagc caaagtaaca
     3061 gtgaagactt caagctcagt ggacccagca atagcagcat ggaagatttc actaaacgta
     3121 gcactcagag tgaggacttc tccaggcggc acacggtgcc agatagacac atacctcttg
     3181 ctttgccacg acaaaatagt actgggcagg cccagatccg aaaagtggac cagggtgggt
     3241 taggtgcccg agccaaagcc ccaccatccc tgccacacag tgcttcttta cgtagcaccg
     3301 ggagcatgtc agtggtgtcc gcagccctgg tggccgaacc tgtgcagaat gggagccggt
     3361 cccggcagca gtcctcttcc tccagagaga gccctgttcc caaagttaga gcaatccaga
     3421 gacaacagac acagcaggtt cagtcacctg tggactctgc cacaatgtcc ccagtagaga
     3481 ggacagcagc ctgggttctg aacaatgggc agtatgaaga ggatgtggaa gaaactgagc
     3541 aaaatctaga tgaagccaag catgctgaga agtatgaaca agaaattact aaactgaagg
     3601 agcgcctgag agtttccagc cggcgactgg aggaatatga acgccgcttg ctggtgcagg
     3661 agcagcagat gcagaagctg ctgctggaat acaaggcccg actggaggac agcgaggagc
     3721 ggctccgaag acagcaggaa gaaaaagata gccagatgaa aagcatcatc agcaggctaa
     3781 tggctgtgga agaggaactg aagaaggatc atgctgagat gcaagcagtt attgatgcaa
     3841 agcagaaaat aattgatgca caggaaaaac ggatcgtgtc cctggattca gccaacacca
     3901 gactgatgag cgcgctgacc caagtgaagg agcggtacag catgcaggtc cgcaatggca
     3961 tctcccccac caaccccacc aagctttcca tcacggagaa tggtgaattc aaaaacagca
     4021 gctgctaatc aactttcttg tacaaagttg gcattatagg aaagcattgc ttatcaattt
     4081 gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgcatg caagcttaga
     4141 tcctctagac cagccaggac agaaatgcct cgacttcgct gctacccaag gttgccgggt
     4201 gacgcacacc gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc
     4261 gtaagctgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac
     4321 gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg
     4381 tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga
     4441 tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca
     4501 ttatgaggga agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca
     4561 tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg
     4621 gcggcctgaa gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg
     4681 aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga
     4741 gcgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc
     4801 gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag
     4861 gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag
     4921 aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac
     4981 aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg
     5041 ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg
     5101 gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt
     5161 atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg
     5221 cctcgcgcgc agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg
     5281 tagtcggcaa ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt
     5341 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5401 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5461 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5521 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5581 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5641 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5701 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5761 gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5821 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5881 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5941 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6001 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     6061 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     6121 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     6181 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     6241 cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa
     6301 cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg
     6361 cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg
     6421 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
     6481 gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact ctcgcg
//
