LOCUS       sequence-125961-        5831 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway ORF clone of human RASA1 [NM_002890.2] without stop codon 
            (for C-terminal fusions).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5831)
  TITLE     Esposito lab
REFERENCE   2  (bases 1 to 5831)
  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-E228 Hs.RASA1-nostop
FEATURES             Location/Qualifiers
     source          1..5831
                     /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(3322..3421)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3798..4589
                     /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      4682..5270
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5171..5190
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5424..5441
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5600..5627)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5719..5805)
                     /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 aggtcaactt tcttgtacaa agttggcatt ataggaaagc
     3361 attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt
     3421 gcatgcaagc ttagatcctc tagaccagcc aggacagaaa tgcctcgact tcgctgctac
     3481 ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc cagtggacat
     3541 aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc aactggtcca
     3601 gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg
     3661 actgtttttt tggggtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg
     3721 tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagggc agtcgcccta
     3781 aaacaaagtt aaacattatg agggaagcgg tgatcgccga agtatcgact caactatcag
     3841 aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt gctggccgta catttgtacg
     3901 gctccgcagt ggatggcggc ctgaagccac acagtgatat tgatttgctg gttacggtga
     3961 ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg
     4021 cttcccctgg agagagcgag attctccgcg ctgtagaagt caccattgtt gtgcacgacg
     4081 acatcattcc gtggcgttat ccagctaagc gcgaactgca atttggagaa tggcagcgca
     4141 atgacattct tgcaggtatc ttcgagccag ccacgatcga cattgatctg gctatcttgc
     4201 tgacaaaagc aagagaacat agcgttgcct tggtaggtcc agcggcggag gaactctttg
     4261 atccggttcc tgaacaggat ctatttgagg cgctaaatga aaccttaacg ctatggaact
     4321 cgccgcccga ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt
     4381 acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg gcaatggagc
     4441 gcctgccggc ccagtatcag cccgtcatac ttgaagctag acaggcttat cttggacaag
     4501 aagaagatcg cttggcctcg cgcgcagatc agttggaaga atttgtccac tacgtgaaag
     4561 gcgagatcac caaggtagtc ggcaaataac cctcgagcca cccatgacca aaatccctta
     4621 acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4681 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4741 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4801 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     4861 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4921 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4981 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5041 cctacaccga actgagatac ctacagcgtg agcattgaga aagcgccacg cttcccgaag
     5101 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5161 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5221 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5281 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     5341 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     5401 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     5461 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     5521 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc gctagccagg
     5581 aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct tagtttgatg
     5641 cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt
     5701 caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat
     5761 aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc tggcagttcc
     5821 ctactctcgc g
//
