LOCUS       sequence-117495-        5755 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tol1 expression plasmid with UAS-driven myc tagged nlsTdTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5755)
  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 5755)
  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-nlstdTomato
FEATURES             Location/Qualifiers
     source          1..5755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    628..648
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     650..669
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        650..668
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     693..709
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     694..710
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1060..1076)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1060..1076)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(1113..1131)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    1131..1155
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1158..1167
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1194..1223
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1233..1262
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1272..1301
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1311..1340
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1350..1379
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1413..1442
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1467..1487
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1488..2915
                     /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
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     CDS             2928..2984
                     /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"
     protein_bind    complement(2987..3011)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3037..3057)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     polyA_signal    3079..3213
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3103..3122)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3157..3176
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3482..3498)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3534..3550)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3547..3569)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3558..3574
                     /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(3582..3588,3589..3606,3607..3612))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3627..3648
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3765..3782)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3936..4524)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4016..4035)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4695..5486,5487..5555))
                     /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     5318..5337
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5556..5660)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5728..5746
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt ccctttagcc agtagcggtt
      421 ctaggcacgg gccgtccggg cggtggcctg gggcggaaaa ctgaaggggg gcggcaccgg
      481 cggctcagcc ctttgtaata tattaatatg caccactatt ggtttactta tgtcacagtt
      541 tgtaagtttg taacagcctg aacctggccg cgcgccattg ggatggaacc ctcgaggggc
      601 ccgctagcga tatccctgca gggctatcaa ctttgtatag aaaagttggt aatacgactc
      661 actatagggc gaattgggta ccgggccccc cctcgaggtc gacggtatcg ataagcttag
      721 gcctccaagg cggagtactg tcctccgggc tggcggagta ctgtcctccg gcaaggtcgg
      781 agtactgtcc tccgacacta gaggtcggag tactgtcctc cgacgcaagg cggagtactg
      841 tcctccgggc tgcggagtac tgtcctccgg caaggtcgga gtactgtcct ccgacactag
      901 aggtcggagt actgtcctcc gacgcaaggc ggagtactgt cctccgggct ggcggagtac
      961 tgtcctccgg caagggtcga ctctagaggg tatataatgg atcccatcgc gtctcagcct
     1021 cactttgagc tcctccacac gaattcctgc agcccggggg atccactagt tctagagcgg
     1081 ccgccaccgc ggtggagctc cagcttttgt tccctttagt gagggttaat tcaagtttgt
     1141 acaaaaaagc aggctccgcc accatggctg gatcccatcg atttaaagct atggagcaaa
     1201 agctcatttc tgaagaggac ttgaatgaaa tggagcaaaa gctcatttct gaagaggact
     1261 tgaatgaaat ggagcaaaag ctcatttctg aagaggactt gaatgaaatg gagcaaaagc
     1321 tcatttctga agaggacttg aatgaaatgg agcaaaagct catttctgaa gaggacttga
     1381 atgaaatgga gagcttgggc gacctcacca tggagcaaaa gctcatttct gaagaggact
     1441 tgaatttaga gatctcgaat tccatgccca agaagaagag gaaggtgatg gtgagcaagg
     1501 gcgaggaggt catcaaagag ttcatgcgct tcaaggtgcg catggagggc tccatgaacg
     1561 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     1621 ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc
     1681 agttcatgta cggctccaag gcgtacgtga agcaccccgc cgacatcccc gattacaaga
     1741 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggtc
     1801 tggtgaccgt gacccaggac tcctccctgc aggacggcac gctgatctac aaggtgaaga
     1861 tgcgcggcac caacttcccc cccgacggcc ccgtaatgca gaagaagacc atgggctggg
     1921 aggcctccac cgagcgcctg tacccccgcg acggcgtgct gaagggcgag atccaccagg
     1981 ccctgaagct gaaggacggc ggccactacc tggtggagtt caagaccatc tacatggcca
     2041 agaagcccgt gcaactgccc ggctactact acgtggacac caagctggac atcacctccc
     2101 acaacgagga ctacaccatc gtggaacagt acgagcgctc cgagggccgc caccacctgt
     2161 tcctggggca tggcaccggc agcaccggca gcggcagctc cggcaccgcc tcctccgagg
     2221 acaacaacat ggccgtcatc aaagagttca tgcgcttcaa ggtgcgcatg gagggctcca
     2281 tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc
     2341 agaccgccaa gctgaaggtg accaagggcg gccccctgcc cttcgcctgg gacatcctgt
     2401 ccccccagtt catgtacggc tccaaggcgt acgtgaagca ccccgccgac atccccgatt
     2461 acaagaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg
     2521 gcggtctggt gaccgtgacc caggactcct ccctgcagga cggcacgctg atctacaagg
     2581 tgaagatgcg cggcaccaac ttcccccccg acggccccgt aatgcagaag aagaccatgg
     2641 gctgggaggc ctccaccgag cgcctgtacc cccgcgacgg cgtgctgaag ggcgagatcc
     2701 accaggccct gaagctgaag gacggcggcc actacctggt ggagttcaag accatctaca
     2761 tggccaagaa gcccgtgcaa ctgcccggct actactacgt ggacaccaag ctggacatca
     2821 cctcccacaa cgaggactac accatcgtgg aacagtacga gcgctccgag ggccgccacc
     2881 acctgttcct gtacggcatg gacgagctgt acaagggagg atccggagcc acgaacttct
     2941 ctctgttaaa gcaagcagga gacgtggaag aaaaccccgg tcctccaccc agctttcttg
     3001 tacaaagtgg ggcgcgccta cgtaactagt ctcgagcaac tttattatac atagttgata
     3061 aggcgcgcca ggcctgtaga tccagacatg ataagataca ttgatgagtt tggacaaacc
     3121 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     3181 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     3241 tttcaggttc agggggaggt gtgggaggtt ttttaattaa aacatttcag ttgacgaaga
     3301 caaacaaagt tctgttgtga ctatgggggg ggggggcgcc tggggatggt ctcgcccggg
     3361 gagtaattca gggtagaacc gccactggct aaagggagct cggtacctcg cgaatgcatc
     3421 tagatatcgg atcccgggcc cgtcgactgc agaggcctgc atgcaagctt gcgtaatcat
     3481 ggtcatagct gtttcctgta tcccaatggc gcgccgagct tggcgtaatc atggtcatag
     3541 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     3601 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     3661 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3721 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     3781 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3841 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3901 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3961 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4021 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4081 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4141 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4201 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4261 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4321 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4381 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4441 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4501 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4561 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4621 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4681 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4741 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4801 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4861 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4921 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4981 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5041 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5101 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5161 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5221 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5281 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5341 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5401 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5461 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5521 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5581 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5641 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     5701 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtc
//
