LOCUS       sequence-201387-        6869 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  UAS-driven expression of activated MEK.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6869)
  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 6869)
  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: pTol1:uas-myc-act-map2k2b-2a-tdTomato
FEATURES             Location/Qualifiers
     source          1..6869
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(42..64)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     164..183
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     377..399
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     391..408
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     392..408
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    641..661
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        663..681
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     706..722
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     707..723
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1073..1089)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1073..1089)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(1126..1144)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    1144..1168
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1213..1242
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1252..1281
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1291..1320
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1330..1359
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1369..1398
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1432..1461
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    complement(2708..2732)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2743..2799
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2800..4230
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGSHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     protein_bind    complement(4232..4252)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     polyA_signal    4274..4408
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4298..4317)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4352..4371
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4609..4625)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4661..4677)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4674..4696)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4685..4701
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4709..4715,4716..4733,4734..4739))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4754..4775
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4892..4909)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5063..5651)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5143..5162)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5822..6613,6614..6682))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6445..6464
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6683..6787)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(6855..6869,1..4)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
       61 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
      121 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
      181 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
      241 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
      301 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
      361 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attcccttta
      421 gccagtagcg gttctaggca cgggccgtcc gggcggtggc ctggggcgga aaactgaagg
      481 ggggcggcac cggcggctca gccctttgta atatattaat atgcaccact attggtttac
      541 ttatgtcaca gtttgtaagt ttgtaacagc ctgaacctgg ccgcgcgcca ttgggatgga
      601 accctcgagg ggcccgctag cgatatccct gcagggctat caactttgta tagaaaagtt
      661 ggtaatacga ctcactatag ggcgaattgg gtaccgggcc ccccctcgag gtcgacggta
      721 tcgataagct taggcctcca aggcggagta ctgtcctccg ggctggcgga gtactgtcct
      781 ccggcaaggt cggagtactg tcctccgaca ctagaggtcg gagtactgtc ctccgacgca
      841 aggcggagta ctgtcctccg ggctgcggag tactgtcctc cggcaaggtc ggagtactgt
      901 cctccgacac tagaggtcgg agtactgtcc tccgacgcaa ggcggagtac tgtcctccgg
      961 gctggcggag tactgtcctc cggcaagggt cgactctaga gggtatataa tggatcccat
     1021 cgcgtctcag cctcactttg agctcctcca cacgaattcc tgcagcccgg gggatccact
     1081 agttctagag cggccgccac cgcggtggag ctccagcttt tgttcccttt agtgagggtt
     1141 aattcaagtt tgtacaaaaa agcaggctcc gccaccatgc tttttgcagg atcccatcga
     1201 tttaaagcta tggagcaaaa gctcatttct gaagaggact tgaatgaaat ggagcaaaag
     1261 ctcatttctg aagaggactt gaatgaaatg gagcaaaagc tcatttctga agaggacttg
     1321 aatgaaatgg agcaaaagct catttctgaa gaggacttga atgaaatgga gcaaaagctc
     1381 atttctgaag aggacttgaa tgaaatggag agcttgggcg acctcaccat ggagcaaaag
     1441 ctcatttctg aagaggactt gaatttagag atctcgaatt caaggcctct cgagatggct
     1501 cccaaaaaga ggccagatcc tatagttata actggaagcg gagaagcaca gacgacctcc
     1561 acgcccatag atgctgcatc agaagcgaat cttgaagccc tgcagaagaa gctggatgag
     1621 ctggatttgg acgagcagca gaagaagcgt ctggaagctt tcctcacgca gaaagccaat
     1681 ctgggagagc tgaaggacga tgatttccag catatctgtg aactgggagc tggaaacgga
     1741 ggagtggtga ataaagtgtg ccacaaacct tcaggtctcg tcatggcccg gaagttaatt
     1801 cacctggaga tcaagccggc catcaggcat cagatcatca gagagctgca ggtcttgcat
     1861 gagtgtaatt caccatacat cgtgggtttt tacggcgcat tttacagcga tggcgagatt
     1921 agtatctgta tggagaacat ggacggcgga tcgttggatc aggtcctgaa agaggctcgc
     1981 agaatcccag aggagatttt gggaaaagtc agcatagctg ttttaagagg tcttgcttac
     2041 cttcgagaaa agcaccagat tattcacaga gatgtgaaac cctcaaatat tctggtaaac
     2101 tgtcgcgggg agatcaagct gtgtgacttc ggtgtgagcg ggcagctgat cgacgacatg
     2161 gccaacgact ttgtaggaac acgctcatac atgtcgccgg agcggttgca gggcactcat
     2221 tactcggttc agtcagacgt gtggagtatg ggtctctctc tggtggagct cgccatcggc
     2281 cgatacccta ttcctccacc tgacgccaag gagctggagg gcatcttcgg cagggcactg
     2341 atggatgttg gagattcaga aacacacagc acgtctcctc gacccagacc acctggcaga
     2401 cccatcagcg gtcatgggcc ggtcatggct atatttgagc tgctggatta tatagtcaat
     2461 gagcctcctc ctaaactgcc tcacggcgtc ttcactcctg atttccagga ctttgtgacc
     2521 aactgtctta ttaaaaaccc tgcagacaga gcggacctga agatgctgat gaatcacaca
     2581 tttatcaaac gcgccgaagt ggaggagatg gactttgctg gatggttgtg taaaacaatg
     2641 ggtctcaatc agcccagcac acccacacaa acaactgaat ctagaactat agtgagtcgt
     2701 attacgcacc cagctttctt gtacaaagtg gacggatccg gagccacgaa cttctctctg
     2761 ttaaagcaag caggagacgt ggaagaaaac cccggtccta tggtgagcaa gggcgaggag
     2821 gtcatcaaag agttcatgcg cttcaaggtg cgcatggagg gctccatgaa cggccacgag
     2881 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     2941 aaggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc ccagttcatg
     3001 tacggctcca aggcgtacgt gaagcacccc gccgacatcc ccgattacaa gaagctgtcc
     3061 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg tctggtgacc
     3121 gtgacccagg actcctccct gcaggacggc acgctgatct acaaggtgaa gatgcgcggc
     3181 accaacttcc cccccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     3241 accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg agatccacca ggccctgaag
     3301 ctgaaggacg gcagccacta cctggtggag ttcaagacca tctacatggc caagaagccc
     3361 gtgcaactgc ccggctacta ctacgtggac accaagctgg acatcacctc ccacaacgag
     3421 gactacacca tcgtggaaca gtacgagcgc tccgagggcc gccaccacct gttcctgggg
     3481 catggcaccg gcagcaccgg cagcggcagc tccggcaccg cctcctccga ggacaacaac
     3541 atggccgtca tcaaagagtt catgcgcttc aaggtgcgca tggagggctc catgaacggc
     3601 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     3661 aagctgaagg tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag
     3721 ttcatgtacg gctccaaggc gtacgtgaag caccccgccg acatccccga ttacaagaag
     3781 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggtctg
     3841 gtgaccgtga cccaggactc ctccctgcag gacggcacgc tgatctacaa ggtgaagatg
     3901 cgcggcacca acttcccccc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     3961 gcctccaccg agcgcctgta cccccgcgac ggcgtgctga agggcgagat ccaccaggcc
     4021 ctgaagctga aggacggcgg ccactacctg gtggagttca agaccatcta catggccaag
     4081 aagcccgtgc aactgcccgg ctactactac gtggacacca agctggacat cacctcccac
     4141 aacgaggact acaccatcgt ggaacagtac gagcgctccg agggccgcca ccacctgttc
     4201 ctgtacggca tggacgagct gtacaagtaa ccaactttat tatacatagt tgataaggcg
     4261 cgccaggcct gtagatccag acatgataag atacattgat gagtttggac aaaccacaac
     4321 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     4381 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     4441 ggttcagggg gaggtgtggg aggtttttta attaaaacat ttcagttgac gaagacaaac
     4501 aaagttctgt tgtgactatg gggggggggg gcgcctgggg atggtctcgc ccggggagta
     4561 attcagggta gaaccgccac tggctaaagg aagcttggcg taatcatggt catagctgtt
     4621 tcctgtatcc caatggcgcg ccgagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     4681 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     4741 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     4801 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     4861 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4921 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4981 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5041 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5101 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5161 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5221 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5281 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5341 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5401 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5461 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5521 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5581 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5641 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5701 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5761 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5821 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5881 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5941 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6001 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6061 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6121 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6181 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6241 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6301 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6361 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6421 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6481 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6541 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6601 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6661 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6721 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6781 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     6841 cattaaccta taaaaatagg cgtatcacg
//
