LOCUS       sequence-396359-        3141 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway middle entry vector expressing constitutively actve 
            zebrafish rras.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3141)
  AUTHORS   Greysson-Wong J, Rode R, Ryu JR, Chan JL, Davari P, Rinker KD, 
            Childs SJ
  TITLE     rasa1-related arteriovenous malformation is driven by aberrant 
            venous signalling.
  JOURNAL   Development. 2023 Sep 15;150(18):dev201820. doi: 10.1242/dev.201820.
            Epub 2023 Sep 14.
  PUBMED    37708300
REFERENCE   2  (bases 1 to 3141)
  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"; journalName: 
            "Development."; date: "2023-09-15"; volume: "150(18):dev201820. 
            doi"; pages: " 10.1242/dev.201820. Epub 2023 Sep 14"
COMMENT     Sequence Label: pME rras_Q61R
FEATURES             Location/Qualifiers
     source          1..3141
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1266..1365)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1382..1401)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1383..1401)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1406..1422)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1406..1422)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1524..1543)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1535..2344
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1602..1621)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2491..3079
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2980..2999
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggcttc accatgtcaa cggaagaaag atacaagttg gtggttgtgg gcggtggagg
      721 tgtggggaaa agcgccctca ccattcagtt catccagtcg tactttgtgt ctgactacga
      781 cccaaccatt gaagactctt acactaaaat ctgcaccgtg gacgggaagg agacacgatt
      841 agacatcctc gacacagcag ggcgagagga gtttggagcc atgagagagc agtacatgcg
      901 ctctggagag ggcttcttac tggtgtttgc tcttaatgac agtggcagtt ataacgaaat
      961 tcagaaattt cacactcaga tattacgggt taaggatcga gatgactttc ctatggttct
     1021 tgttggcaac aaatcagatc ttgaccagca gagagtgatc tcaaaagagg aagccatgac
     1081 gtttgcccga gaaaacagga tccactatat ggaatcatca gccaagaacc gtcacaacgt
     1141 ggatgaagcc ttcatggagg ttgtgcgagc aataaggaag tttcaggaga cagagagtcc
     1201 accgcttccc gctaatcata cagggaagca gaagagtggt ggctgtccct gcaccctact
     1261 ctgaaaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     1321 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     1381 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     1441 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     1501 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     1561 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     1621 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     1681 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     1741 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     1801 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     1861 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     1921 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     1981 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     2041 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     2101 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     2161 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     2221 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     2281 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     2341 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     2401 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     2461 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2521 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2581 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     2641 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2701 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2761 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2821 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     2881 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     2941 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3001 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3061 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3121 ctggcctttt gctcacatgt t
//
