LOCUS       sequence-125960-        5834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway ORF clone of human RASA1 [NM_002890.2] 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 5834)
  TITLE     Esposito lab
REFERENCE   2  (bases 1 to 5834)
  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-E227 Hs.RASA1
FEATURES             Location/Qualifiers
     source          1..5834
                     /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"
     regulatory      184..193
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3325..3424)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3801..4592
                     /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      4685..5273
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5174..5193
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5427..5444
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5603..5630)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5722..5808)
                     /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 ggccgaggcc ggcagtgagg agggcggccc ggtaacagcc ggagctggag
      241 gaggcggcgc ggcagcgggc tccagtgcct atcccgcagt gtgtcgggtg aagatacccg
      301 cggccctgcc tgtggcagcc gccccctatc ctgggctggt ggagaccgga gtggctggaa
      361 ctctgggtgg cggagccgct ttggggtcag agttcctagg agccgggtct gtggcagggg
      421 cactgggggg agctggactg acagggggag gtactgctgc tggcgtagct ggtgctgctg
      481 ctggcgtggc cggtgctgct gttgctggac ctagtggaga catggctctc accaaactgc
      541 ccacttcgtt gcttgctgag actctcgggc caggcggcgg ttttccccct ctgccccctc
      601 ccccttacct gccccctttg ggggcgggcc tcgggacagt ggacgaaggt gactctctgg
      661 atggaccaga atacgaggag gaagaggtgg ccataccgtt gaccgctcct ccaactaacc
      721 agtggtatca cggaaaactt gacagaacga tagcagaaga acgcctcagg caggcaggga
      781 agtctggcag ttatcttata agagagagtg atcggaggcc agggtccttt gtactttcat
      841 ttcttagcca gatgaatgtt gtcaaccatt ttaggattat tgctatgtgt ggagattact
      901 acattggtgg aagacgtttt tcttcactgt cagacctaat aggttattac agtcatgttt
      961 cttgtttgct taaaggagaa aaattacttt acccagttgc accaccagag ccagtagaag
     1021 atagaaggcg tgtacgagct attctacctt acacaaaagt accagacact gatgaaataa
     1081 gtttcttaaa aggagatatg ttcattgttc ataatgaatt agaagatgga tggatgtggg
     1141 ttacaaattt aagaacagat gaacaaggcc ttattgttga agacctagta gaagaggtgg
     1201 gccgggaaga agatccacat gaaggaaaaa tatggttcca tgggaagatt tccaaacagg
     1261 aagcttataa tttactaatg acagttggtc aagtctgcag ttttcttgtg aggccctcag
     1321 ataatactcc tggcgattat tcactttatt tccggaccaa tgaaaatatt cagcgattta
     1381 aaatatgtcc aacgccaaac aatcagttta tgatgggagg ccggtattat aacagcattg
     1441 gggacatcat agatcactat cgaaaagaac agattgttga aggatattat cttaaggaac
     1501 ctgtaccaat gcaggatcaa gaacaagtac tcaatgacac agtggatggc aaggaaatct
     1561 ataataccat ccgtcgtaaa acaaaggatg ccttttataa aaacattgtt aagaaaggtt
     1621 atcttctgaa aaagggcaaa ggaaaacgtt ggaaaaattt atattttatc ttagagggta
     1681 gtgatgccca acttatttat tttgaaagcg aaaaacgagc taccaaacca aaaggattaa
     1741 tagatctcag tgtatgttct gtctatgtcg ttcatgatag tctctttggc aggccaaact
     1801 gttttcagat agtagttcag cactttagtg aagaacatta catcttttac tttgcaggag
     1861 aaactccaga acaagcagag gattggatga aaggtctgca ggcattttgc aatttacgga
     1921 aaagtagtcc agggacatcc aataaacgcc ttcgtcaggt cagcagcctt gttttacata
     1981 ttgaagaagc ccataaactc ccagtaaaac attttactaa tccatattgt aacatctacc
     2041 tgaatagtgt ccaagtagca aaaactcatg caagggaagg gcaaaaccca gtatggtcag
     2101 aagagtttgt ctttgatgat cttcctcctg acatcaatag atttgaaata actcttagta
     2161 ataaaacaaa gaaaagcaaa gatcctgata tcttatttat gcgctgccag ttgagccgat
     2221 tacagaaagg gcatgccaca gatgaatggt ttctgctcag ctcccatata ccattaaaag
     2281 gtattgaacc agggtccctg cgtgttcgag cacgatactc tatggaaaaa atcatgccag
     2341 aagaagagta cagtgaattt aaagagctta tactgcaaaa ggaacttcat gtagtctatg
     2401 ctttatcaca tgtatgtgga caagaccgaa cactactggc cagcatccta ctgaggattt
     2461 ttcttcacga aaagcttgaa tcgttgttgt tatgcacact aaatgacaga gaaataagca
     2521 tggaagatga agccactacc ctatttcgag ccacaacact tgcaagcacc ttgatggagc
     2581 agtatatgaa agccactgct acacagtttg ttcatcatgc tttgaaagac tctattttaa
     2641 agataatgga aagcaagcag tcttgtgagt taagtccatc aaagttagaa aaaaatgaag
     2701 atgtgaacac taatttaaca cacctattga acatactttc agagcttgtg gagaaaatat
     2761 tcatggcttc agaaatactt ccaccgacat tgagatatat ttatgggtgt ttacagaaat
     2821 ctgttcagca taagtggcct acaaatacca ccatgagaac aagagttgtt agtggttttg
     2881 tttttcttcg actcatctgt cctgccatcc tgaatccacg gatgttcaat atcatctcag
     2941 attctccatc tcctattgct gcaagaacac tgatattagt ggctaaatct gtgcagaact
     3001 tagcaaatct tgtggaattt ggagctaagg agccctacat ggaaggtgtc aatccattca
     3061 tcaaaagcaa caaacatcgt atgatcatgt ttttagatga acttgggaat gtacctgaac
     3121 ttccggacac tacagagcat tctagaacgg acctgtcccg tgatttagca gcattgcatg
     3181 agatttgcgt ggctcattca gatgaacttc gaacgctcag taatgagcgt ggtgcacagc
     3241 agcacgtatt gaaaaagctt ctggctataa cagaactgct tcaacaaaaa caaaaccagt
     3301 atacaaaaac caatgatgtc aggtaatcaa ctttcttgta caaagttggc attataggaa
     3361 agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta
     3421 tttgcatgca agcttagatc ctctagacca gccaggacag aaatgcctcg acttcgctgc
     3481 tacccaaggt tgccgggtga cgcacaccgt ggaaacggat gaaggcacga acccagtgga
     3541 cataagcctg ttcggttcgt aagctgtaat gcaagtagcg tatgcgctca cgcaactggt
     3601 ccagaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc atggcttgtt
     3661 atgactgttt ttttggggta cagtctatgc ctcgggcatc caagcagcaa gcgcgttacg
     3721 ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt tacgcagcag ggcagtcgcc
     3781 ctaaaacaaa gttaaacatt atgagggaag cggtgatcgc cgaagtatcg actcaactat
     3841 cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc gtacatttgt
     3901 acggctccgc agtggatggc ggcctgaagc cacacagtga tattgatttg ctggttacgg
     3961 tgaccgtaag gcttgatgaa acaacgcggc gagctttgat caacgacctt ttggaaactt
     4021 cggcttcccc tggagagagc gagattctcc gcgctgtaga agtcaccatt gttgtgcacg
     4081 acgacatcat tccgtggcgt tatccagcta agcgcgaact gcaatttgga gaatggcagc
     4141 gcaatgacat tcttgcaggt atcttcgagc cagccacgat cgacattgat ctggctatct
     4201 tgctgacaaa agcaagagaa catagcgttg ccttggtagg tccagcggcg gaggaactct
     4261 ttgatccggt tcctgaacag gatctatttg aggcgctaaa tgaaacctta acgctatgga
     4321 actcgccgcc cgactgggct ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt
     4381 ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg
     4441 agcgcctgcc ggcccagtat cagcccgtca tacttgaagc tagacaggct tatcttggac
     4501 aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc cactacgtga
     4561 aaggcgagat caccaaggta gtcggcaaat aaccctcgag ccacccatga ccaaaatccc
     4621 ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4681 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4741 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4801 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     4861 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4921 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4981 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5041 cgacctacac cgaactgaga tacctacagc gtgagcattg agaaagcgcc acgcttcccg
     5101 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5161 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5221 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5281 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     5341 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     5401 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     5461 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     5521 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt accgctagcc
     5581 aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg
     5641 atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac
     5701 gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
     5761 gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat gcctggcagt
     5821 tccctactct cgcg
//
