LOCUS       sequence-396366-        3743 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway middle entry vector expressing constitutively active 
            zebrafish map2k2a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3743)
  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 3743)
  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 map2k2a_s219D
FEATURES             Location/Qualifiers
     source          1..3743
                     /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(1868..1967)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1984..2003)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1985..2003)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2008..2024)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2008..2024)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2126..2145)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2137..2946
                     /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(2204..2223)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3093..3681
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3582..3601
                     /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 agcaggctcc accatggcac caaaaagaag accagtcccc ttgattataa cccccactgg
      721 agagggacag tcaaccaaca ttgatgcagc agcagaggcc aacctggagg ccttgcagag
      781 gaagttaggt gagctggatt tggatgaaca gcagaggaag cgcctcgaag ctttcctcac
      841 ccagaaagcc caggtgggcg agctcaagga tgaagatttc gaacccatat gtgaacttgg
      901 cgctggtaat ggtggagtcg ttcataaagt ccgtcacaaa ccctcgagac tggttatggc
      961 caggaagctt atacacttgg aaatcaaacc agccatcagg aaccagatta ttcgagagct
     1021 gcaggttcta cacgagtgta actcaccgta cattgtaggg ttttacggcg ccttctacag
     1081 tgatggagaa atcagcatct gtatggagca catggatgga ggctctttgg atcaagtgct
     1141 caaagaagcc agaagaatcc ctgaggaaat tttgggcaaa gttagcatag ctgtactcag
     1201 aggtttggcg tatcttcggg agaagcacca aataatgcac agagacgtta agccctccaa
     1261 catcctggtg aactcgcgcg gtgagatcaa actgtgcgac tttggtgtga gtggccagct
     1321 catcgacgat atggccaact ccttcgttgg aacacggtcg tacatgtcgc cggagagact
     1381 ccagggcacg cactactcgg ttcagtcgga tgtgtggagc atgggcctat cactggttga
     1441 gcttgctatt ggacgctacc ccatcccccc tcccgatgct aaggaactgg aggccatatt
     1501 tggccggcca gtgctggatg caggtggggc agaaggtcac agcatgtctc caagacccag
     1561 gcctccagga cgaccagtta gtggacacgg aatggacagc aggccagcaa tggctatatt
     1621 tgagctactg gactacattg tcaatgagcc tccacccaag ctcccccatg gcgtcttcac
     1681 cacagacttt gaggagtttg tgacaaaatg cctcataaag aaccctgctg acagagctga
     1741 cctcaagatg ctaatgggtc acacgttcat caagcgggct gaggttgagg aggtggattt
     1801 tgctggatgg ttgtgtaaaa ccatgggcct tcaccagcct agcacaccca ctcatagcgc
     1861 tgaatgaacc cagctttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt
     1921 tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat
     1981 atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg
     2041 tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa
     2101 ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg
     2161 tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
     2221 gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca
     2281 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     2341 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
     2401 cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta
     2461 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
     2521 ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct
     2581 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     2641 aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg
     2701 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     2761 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     2821 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     2881 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     2941 ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg
     3001 gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg
     3061 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3121 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3181 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     3241 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3301 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3361 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3421 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3481 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3541 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3601 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3661 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     3721 tgctggcctt ttgctcacat gtt
//
