LOCUS       sequence-117329-        4139 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3' entry 2A-tdTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4139)
  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 4139)
  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: p3E-2a-tdTomato
FEATURES             Location/Qualifiers
     source          1..4139
                     /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    complement(624..747)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             759..815
                     /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             816..2246
                     /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(2248..2343)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2380..2399)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2381..2399)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2404..2420)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2404..2420)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2522..2541)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2533..3342
                     /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(2600..2619)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3489..4077
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3978..3997
                     /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 tcgggccctg cagctctaga gctcgaattc tacaggtcac
      601 taataccatc taagtagttg attcatagtg actgcatatg ttgtgtttta cagtattatg
      661 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt
      721 ctcgttcaac tttcttgtac aaagtggacg gatccggagc cacgaacttc tctctgttaa
      781 agcaagcagg agacgtggaa gaaaaccccg gtcctatggt gagcaagggc gaggaggtca
      841 tcaaagagtt catgcgcttc aaggtgcgca tggagggctc catgaacggc cacgagttcg
      901 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
      961 tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag ttcatgtacg
     1021 gctccaaggc gtacgtgaag caccccgccg acatccccga ttacaagaag ctgtccttcc
     1081 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggtctg gtgaccgtga
     1141 cccaggactc ctccctgcag gacggcacgc tgatctacaa ggtgaagatg cgcggcacca
     1201 acttcccccc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctccaccg
     1261 agcgcctgta cccccgcgac ggcgtgctga agggcgagat ccaccaggcc ctgaagctga
     1321 aggacggcag ccactacctg gtggagttca agaccatcta catggccaag aagcccgtgc
     1381 aactgcccgg ctactactac gtggacacca agctggacat cacctcccac aacgaggact
     1441 acaccatcgt ggaacagtac gagcgctccg agggccgcca ccacctgttc ctggggcatg
     1501 gcaccggcag caccggcagc ggcagctccg gcaccgcctc ctccgaggac aacaacatgg
     1561 ccgtcatcaa agagttcatg cgcttcaagg tgcgcatgga gggctccatg aacggccacg
     1621 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc
     1681 tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc ccccagttca
     1741 tgtacggctc caaggcgtac gtgaagcacc ccgccgacat ccccgattac aagaagctgt
     1801 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggtctggtga
     1861 ccgtgaccca ggactcctcc ctgcaggacg gcacgctgat ctacaaggtg aagatgcgcg
     1921 gcaccaactt cccccccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct
     1981 ccaccgagcg cctgtacccc cgcgacggcg tgctgaaggg cgagatccac caggccctga
     2041 agctgaagga cggcggccac tacctggtgg agttcaagac catctacatg gccaagaagc
     2101 ccgtgcaact gcccggctac tactacgtgg acaccaagct ggacatcacc tcccacaacg
     2161 aggactacac catcgtggaa cagtacgagc gctccgaggg ccgccaccac ctgttcctgt
     2221 acggcatgga cgagctgtac aagtaaccaa ctttattata caaagttggc attataaaaa
     2281 agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta
     2341 tttggagctc catggtagcg ttaacgcggc cgcgatatcc cctatagtga gtcgtattac
     2401 atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt
     2461 gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat
     2521 acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac
     2581 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     2641 acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     2701 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     2761 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
     2821 actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     2881 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
     2941 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac
     3001 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     3061 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     3121 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     3181 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     3241 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     3301 aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg
     3361 ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa
     3421 tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3481 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3541 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3601 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     3661 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3721 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3781 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3841 cgaacgacct acaccgaact gagataccta cagcgtgagc attgagaaag cgccacgctt
     3901 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3961 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4021 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4081 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtt
//
