LOCUS       sequence-121121-        4830 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  middle entry plasmid with dimerizable extracellular domain fused to 
            um200 mutant Danio Flt4 cytoplasmic domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4830)
  AUTHORS   Shin M, Male I, Beane TJ, Villefranc JA, Kok FO, Zhu LJ, Lawson ND
  TITLE     Vegfc acts through ERK to induce sprouting and differentiation of 
            trunk lymphatic progenitors.
  JOURNAL   Development. 2016 Oct 15;143(20):3785-3795. Epub 2016 Sep 12.
  PUBMED    27621059
REFERENCE   2  (bases 1 to 4830)
  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: "2016-10-15"; volume: "143(20)"; pages: 
            "3785-3795. Epub 2016 Sep 12"
COMMENT     Sequence Label: pME-Fv2E-flt4um200-ns
FEATURES             Location/Qualifiers
     source          1..4830
                     /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"
     CDS             674..715
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSKPKDPSQR"
     CDS             722..1042
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=DmrB
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             1049..1369
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=DmrB
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             2924..2950
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(2955..3054)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3071..3090)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3072..3090)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3095..3111)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3095..3111)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3213..3232)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3224..4033
                     /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(3291..3310)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4180..4768
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4669..4688
                     /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 agcaggctgg accatgggga gtagcaagag caagcctaag gaccccagcc agcgctctag
      721 aggcgtccaa gtcgaaacca ttagtcccgg cgatggcaga acatttccta aaaggggaca
      781 aacatgtgtc gtccattata caggcatgtt ggaggacggc aaaaaggtgg acagtagtag
      841 agatcgcaat aaacctttca aattcatgtt gggaaaacaa gaagtcatta ggggatggga
      901 ggagggcgtg gctcaaatgt ccgtcggcca acgcgctaag ctcaccatca gccccgacta
      961 cgcatacggc gctaccggac atcccggaat tattccccct cacgctacct tggtgtttga
     1021 cgtcgaactg ttgaagctcg agactagagg agtgcaggtg gagactatct ccccaggaga
     1081 cgggcgcacc ttccccaagc gcggccagac ctgcgtggtg cactacaccg ggatgcttga
     1141 agatggaaag aaagttgatt cctcccggga cagaaacaag ccctttaagt ttatgctagg
     1201 caagcaggag gtgatccgag gctgggaaga aggggttgcc cagatgagtg tgggtcagag
     1261 agccaaactg actatatctc cagattatgc ctatggtgcc actgggcacc caggcatcat
     1321 cccaccacat gccactctcg tcttcgatgt ggagcttcta aaactggaaa ctagagtgaa
     1381 tgtcaagcga gtgaacccag ctgatatcaa gacaggctat ctgtccatca tcatggaccc
     1441 gggggaagtg ccgctggagg agcagtgtga atacctgccg tacgactcca gccagtggga
     1501 gatttccaga gacagactgc gcctcgggaa ggtgctcggc catggagctt tcgggaaggt
     1561 gattgaagcc tcaatctttg gtcatgacaa gaaatcctct gcaaacacag tggctgtcaa
     1621 gatgttgaaa gagggagcaa cagccagcga gcacaaagcc ctgatgtcag agctgaagat
     1681 cctcattcat attggaaacc atctcaatgt tgtcaacctt cttggagctt gtaccaaacc
     1741 taacggtcct ctcatggtta tagtggagta ttgtaaatat ggaaatctgt ccaacttcct
     1801 gcgtgccaag agggagtttt ttttaccata cagggatcgt tctcctaaaa cccaaagtca
     1861 agtgcgacgg atgatagaag caggtcaggc gagccagagt gagcatcagc cgtccacttc
     1921 ctccaccaac cctccacgtg ttacagtgga tgatctctgg aaaactccac tcacaataga
     1981 ggatctgata tgctacagtt ttcaagttgc aagaggaatg gagtttctgg catctcgtaa
     2041 gtgtattcat cgagatctgg cagcccgaaa catcctcctg tccgagagca atgtggtgaa
     2101 gatctgtgac tttggtttag cacgggacat ctacaaagac cctgactatg tgcgcaaagg
     2161 caatgccagg ctgccgctga agtggatggc tccagagagc atctttgata aggtttacac
     2221 cagtcagagt gacgtctggt cttttggagt tctgctctgg gagattttct cactaggagc
     2281 gtctccatac cccggcatcc agattgatga agatttctgc aaacgactca aagacggtac
     2341 caggatgaga gcaccagaca atgcgtcccc tgaaatatat ggcatcatgt tggcctgctg
     2401 gcagggcgag cccagggaga gacccacatt tccagctctg gtggagatac ttggagatct
     2461 actgcaggaa aacagtctac cagaaatccc gttcaatgtg tctcagagct cagaggacga
     2521 tggcttctca caagcttctt ccagaccacc atcacaagag gagatcagat tggcctgcac
     2581 atgtcattct cgagtgaaga cctttgagga acttcctatg gaaatgacat cacacaagac
     2641 acagcatgac agtcagacag acagtgggat ggttttggcg tcagatgaac tggagaggtt
     2701 tgaacacaag cacagaggag ccatgttgac aactgcgacg acgggccaaa gcactgagcg
     2761 cctcatcagt tgtccatcgg tgagcagcag cggcagcggt ggtggtttgc tccgtccagt
     2821 gttcttcact caactctcag gccagacctt ctacaacaac gagtacggtc atctgtcaga
     2881 ggagggcgtc agcgactact tctcctcctc agacccgact agttatccgt acgacgtacc
     2941 agactacgca ttagacccag ctttcttgta caaagttggc attataagaa agcattgctt
     3001 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     3061 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     3121 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaataatat catcatgaac
     3181 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     3241 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     3301 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
     3361 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     3421 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     3481 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     3541 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     3601 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     3661 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     3721 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     3781 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     3841 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     3901 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     3961 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     4021 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     4081 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     4141 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4201 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4261 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4321 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4381 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4441 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4501 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4561 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4621 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4681 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4741 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4801 ggccttttgc tggccttttg ctcacatgtt
//
