LOCUS       sequence-393253-        3009 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 3009)
  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 3009)
  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: 791 p3E-2A-pCS2MCSpA v2
FEATURES             Location/Qualifiers
     source          1..3009
                     /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(61..184)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             196..252
                     /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"
     polyA_signal    334..468
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(358..377)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     412..431
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(587..682)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(715..733)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(738..754)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(738..754)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(856..875)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             867..1676
                     /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(934..953)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1823..2411
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2312..2331
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2565..2582
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2741..2768)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2860..2946)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(2995..3009,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3009,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aaaattctac aggtcactaa taccatctaa gtagttggtt
       61 catagtgact gcatatgttg tgttttacag tattatgtag tctgtttttt atgcaaaatc
      121 taatttaata tattgatatt tatatcattt tacgtttctc gttcaacttt cttgtacaaa
      181 gtggggggat ctggagccac gaacttctct ctgttaaagc aagcaggaga cgtggaagaa
      241 aaccccggtc ctagggatcc catcgattcg aattcaaggc ctctcgagcc tctagtggcg
      301 cgcctctaga actatagtga gtcgtattac gtagatccag acatgataag atacattgat
      361 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
      421 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
      481 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta attcgcggcc
      541 gcggcgccaa tgcattgggc ccggtaccga tccaccggat cttgaccaac tttattatac
      601 aaagttggca ttataaaaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc
      661 agtcaaaata aaatcattat ttggagctcc atggtagcgt taacgcggcc atcccctata
      721 gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc
      781 tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt
      841 acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc
      901 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
      961 ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc
     1021 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact
     1081 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg
     1141 catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc
     1201 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga
     1261 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat
     1321 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc
     1381 ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg
     1441 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
     1501 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga
     1561 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg
     1621 ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag
     1681 aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa
     1741 gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg
     1801 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     1861 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     1921 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     1981 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2041 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     2101 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     2161 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     2221 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     2281 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     2341 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     2401 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     2461 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     2521 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     2581 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     2641 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     2701 atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat
     2761 ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc
     2821 gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc
     2881 gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt
     2941 tatttgatgc ctggcagttc cctactctcg cgttaacgct agcatggatg ttttcccagt
     3001 cacgacgtt
//
