LOCUS       sequence-385190-        3679 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 3679)
  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 3679)
  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: 766 p3E-2A-nlsmCherrypA
FEATURES             Location/Qualifiers
     source          1..3679
                     /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             286..306
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             307..1017
                     /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(452..470)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     720..739
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     914..937
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    1055..1189
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1079..1098)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1133..1152
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(1252..1347)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1385..1403)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1408..1424)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1408..1424)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1526..1545)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1537..2346
                     /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(1604..1623)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2493..3081
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2982..3001
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3235..3252
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3411..3438)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3530..3616)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3665..3679,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3679,1..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 tggctccaaa gaagaagcgt
      301 aaggtaatgg tgagcaaggg cgaggaggac aacatggcca tcatcaagga gttcatgcgc
      361 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
      421 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc
      481 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
      541 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
      601 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
      661 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
      721 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
      781 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
      841 gacgccgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
      901 aacgtcaaca tcaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag
      961 tacgagcgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaaggc
     1021 gcgcctctag aactatagtg agtcgtatta cgtagatcca gacatgataa gatacattga
     1081 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     1141 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     1201 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt ccaactttat
     1261 tatacaaagt tggcattata aaaaagcatt gcttatcaat ttgttgcaac gaacaggtca
     1321 ctatcagtca aaataaaatc attatttgga gctccatggt agcgttaacg cggccgcgat
     1381 atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg
     1441 tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa
     1501 ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg
     1561 tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
     1621 gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca
     1681 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     1741 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
     1801 cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta
     1861 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
     1921 ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct
     1981 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     2041 aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg
     2101 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     2161 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     2221 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     2281 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     2341 ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg
     2401 gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg
     2461 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2521 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     2581 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     2641 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2701 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2761 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2821 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     2881 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     2941 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3001 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3061 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     3121 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     3181 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     3241 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     3301 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     3361 aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat
     3421 ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc
     3481 gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac
     3541 tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag
     3601 cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct agcatggatg
     3661 ttttcccagt cacgacgtt
//
