LOCUS       sequence-385249-        3592 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway p5E vector with reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3592)
  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 3592)
  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: pCK072 p5E 147bp exorh:EGFP
FEATURES             Location/Qualifiers
     source          1..3592
                     /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    57..152
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(350..352,353..355,356..1069)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPIPVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(656..675)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1003..1024
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    1098..1221
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1297..1316)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1298..1316)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1321..1337)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1321..1337)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1439..1458)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1450..2259
                     /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(1517..1536)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2406..2994
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2895..2914
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3148..3165
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3324..3351)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3443..3529)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3578..3592,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3592,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc ccgagttaac gctaccatgg agctccaaat
       61 aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgata agcaatgctt
      121 ttttataatg ccaactttgt atagaaaagt tggctccgaa ttcgccctta gatctggtac
      181 cgcgagcgct gctgtgtctc catctctgga ttatccccct gtctgacccc aatctgacca
      241 ctcactcctg taatctgcct aatctggact catggggttc acataagagc gcagcgccgg
      301 ctctccgggc ttcatctgcg gaggtgtgcc gatcgtccac ttctccatca tggtgagcaa
      361 gggcgaggag ctgttcaccg gggtggtgcc catcccggtc gagctggacg gcgacgtaaa
      421 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
      481 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
      541 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
      601 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
      661 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
      721 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
      781 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
      841 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
      901 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
      961 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1021 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaag gtaccagatc
     1081 taagggcgaa ttcgacccaa gtttgtacaa aaaagttgaa cgagaaacgt aaaatgatat
     1141 aaatatcaat atattaaatt agattttgca taaaaaacag actacataat actgtaaaac
     1201 acaacatatg cagtcactat gaaccaacta cttagatggt attagtgacc tgtagaatta
     1261 attcgagctc tagagctgca gggcggccgc gatatcccct atagtgagtc gtattacatg
     1321 gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca
     1381 caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca
     1441 aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg
     1501 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     1561 atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     1621 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     1681 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     1741 gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     1801 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     1861 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     1921 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     1981 aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc
     2041 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     2101 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     2161 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     2221 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg
     2281 taacactggc agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc
     2341 cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2401 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2461 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2521 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     2581 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2641 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2701 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2761 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2821 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2881 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2941 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3001 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     3061 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     3121 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     3181 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     3241 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc
     3301 caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt
     3361 gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa
     3421 cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac
     3481 agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga tgcctggcag
     3541 ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg tt
//
