LOCUS       sequence-385189-        3652 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway p3E 2A fluorophore.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3652)
  AUTHORS   Kemmler CL, Moran HR, Murray BF, Scoresby A, Klem JR, Eckert RL, 
            Lepovsky E, Bertho S, Nieuwenhuize S, Burger S, D'Agati G, Betz C, 
            Puller AC, Felker A, Ditrychova K, Botschi S, Affolter M, Rohner N, 
            Lovely CB, Kwan KM, Burger A, Mosimann C
  TITLE     Next-generation plasmids for transgenesis in zebrafish and beyond.
  JOURNAL   Development. 2023 Mar 28:dev.201531. doi: 10.1242/dev.201531.
  PUBMED    36975217
REFERENCE   2  (bases 1 to 3652)
  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. 2023 Mar 28:dev.201531. doi: 10.1242/dev.201531."
COMMENT     Sequence Label: 460 p3E-2A-mCherrypA
FEATURES             Location/Qualifiers
     source          1..3652
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(88..211)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             223..279
                     /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             280..990
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(425..443)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     693..712
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     887..910
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    1028..1162
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1052..1071)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1106..1125
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(1225..1320)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1358..1376)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1381..1397)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1381..1397)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1499..1518)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1510..2319
                     /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(1577..1596)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2466..3054
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2955..2974
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3208..3225
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3384..3411)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3503..3589)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3638..3652,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3652,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc cctgcagctc tagagctcga attctacagg
       61 tcactaatac catctaagta gttggttcat agtgactgca tatgttgtgt tttacagtat
      121 tatgtagtct gttttttatg caaaatctaa tttaatatat tgatatttat atcattttac
      181 gtttctcgtt caactttctt gtacaaagtg gggggatccg gagccacgaa cttctctctg
      241 ttaaagcaag caggagacgt ggaagaaaac cccggtccta tggtgagcaa gggcgaggag
      301 gacaacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
      361 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
      421 accgccaagc tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc
      481 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
      541 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
      601 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
      661 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
      721 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
      781 cagaggctga agctgaagga cggcggccac tacgacgccg aggtcaagac cacctacaag
      841 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagct ggacatcacc
      901 tcccacaacg aggactacac catcgtggaa cagtacgagc gcgccgaggg ccgccactcc
      961 accggcggca tggacgagct gtacaagtaa ggcgcgcctc tagaactata gtgagtcgta
     1021 ttacgtagat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     1081 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     1141 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     1201 gggggaggtg tgggaggttt tttccaactt tattatacaa agttggcatt ataaaaaagc
     1261 attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt
     1321 ggagctccat ggtagcgtta acgcggccgc gatatcccct atagtgagtc gtattacatg
     1381 gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca
     1441 caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca
     1501 aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg
     1561 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     1621 atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     1681 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     1741 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     1801 gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     1861 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     1921 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     1981 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     2041 aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc
     2101 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     2161 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     2221 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     2281 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg
     2341 taacactggc agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc
     2401 cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2461 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2521 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2581 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     2641 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2701 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2761 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2821 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2881 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2941 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3001 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3061 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     3121 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     3181 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     3241 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     3301 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc
     3361 caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt
     3421 gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa
     3481 cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac
     3541 agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga tgcctggcag
     3601 ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg tt
//
