LOCUS       sequence-385826-        3284 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway p3E vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3284)
  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 3284)
  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: p3E ubb polyA
FEATURES             Location/Qualifiers
     source          1..3284
                     /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"
     primer_bind     220..236
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     220..236
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(823..839)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(824..840)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    complement(857..952)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(989..1008)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(990..1008)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1013..1029)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1013..1029)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1131..1150)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1142..1951
                     /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(1209..1228)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2098..2686
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2587..2606
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2840..2857
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3016..3043)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3135..3221)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3270..3284,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3284,1..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 ggcggccgct ctagaactag tggatcttct
      241 agaaagattg agtaaggtag aaccgacagt cttagggatg gacattaaat tctcagtatc
      301 ccctgcttat gcactcacca ctttttccca ttattttatt catattgtgt tgctaaatcc
      361 aactacattc gtattaccat gttcttttat tagttttttt ttatctcagc tgtcaaaata
      421 aaagcattga aatacaacat tgttctattt ttcattgtgg ttttgcatgc ccggtaaata
      481 aaaaaacaag gtaatgaaaa taagatgcga aatgagaagt atctcattat gctgtaagaa
      541 gcataattag aatatgcaag tagctttcca aattttaggt gtcttgtctt tctgggcttt
      601 taaagtgttt attcatgtat atttatagat gggaattttt ctgcttactg ctttaaaatg
      661 ttatagcagc tgctgaactt tggtggttct ctttattgtg ttgtatgaac acagaaaatg
      721 gctaaaaatc gctaacactt gatggcaatg aattcatctt gctgaaaaac tcgagccatc
      781 cggaagatcc cccgggctgc aggaattcga tatcaagctt atcgataccg tcgacctcga
      841 gggggggccc ggtacccaac tttattatac aaagttggca ttataaaaaa gcattgctta
      901 tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttggagctcc
      961 atggtagcgt taacgcggcc gcgatatccc ctatagtgag tcgtattaca tggtcatagc
     1021 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     1081 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     1141 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     1201 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     1261 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     1321 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     1381 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     1441 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     1501 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     1561 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     1621 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     1681 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     1741 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     1801 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     1861 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     1921 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     1981 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     2041 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2101 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2161 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2221 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     2281 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2341 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2401 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     2461 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2521 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2581 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2641 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2701 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2761 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2821 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     2881 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     2941 cgactggaaa gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta gccaggaaga
     3001 gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttagt ttgatgcctg
     3061 gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac aacgttcaaa
     3121 tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa
     3181 cgaaaggccc agtcttccga ctgagccttt cgttttattt gatgcctggc agttccctac
     3241 tctcgcgtta acgctagcat ggatgttttc ccagtcacga cgtt
//
