LOCUS       sequence-119505-        2936 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway middle entry vector for full-length attP site for 
            phiC31-mediated transgenesis..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2936)
  AUTHORS   Mosimann C, Puller AC, Lawson KL, Tschopp P, Amsterdam A, Zon LI
  TITLE     Site-directed zebrafish transgenesis into single landing sites with 
            the phiC31 integrase system.
  JOURNAL   Dev Dyn. 2013 Aug;242(8):949-63. doi: 10.1002/dvdy.23989. Epub 2013 
            Jul 3.
  PUBMED    23723152
REFERENCE   2  (bases 1 to 2936)
  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"; doi: 
            "10.1002/dvdy.23989"; journalName: "Dev Dyn"; date: "2013-08"; 
            volume: "242"; issue: "8"; pages: "949-63"
COMMENT     Sequence Label: pENTR5'-attP (pCM254)
FEATURES             Location/Qualifiers
     source          1..2936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    593..688
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    791..890
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     protein_bind    978..1101
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1177..1196)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1178..1196)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1201..1217)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1201..1217)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1319..1338)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1330..2139
                     /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(1397..1416)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2286..2874
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2775..2794
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccga gttaacgcta ccatggagct ccaaataatg
      601 attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgataagca atgctttttt
      661 ataatgccaa ctttgtatag aaaagttggc tccgaattcg cccttcacct ctcgaggtac
      721 ctgcagtact gacggacaca ccgaagcccc ggcggcaacc ctcagcggat gccccggggc
      781 ttcacgtttt cccaggtcag aagcggtttt cgggagtagt gccccaactg gggtaacctt
      841 tgagttctct cagttggggg cgtagggtcg ccgacatgac acaaggggtt gtgaccgggg
      901 tggacacgta cgcgggtgct tacgaccgtc agtcgcgcga gcgcgatcta gagtcgtcga
      961 caagggcgaa ttcgacccaa gtttgtacaa aaaagttgaa cgagaaacgt aaaatgatat
     1021 aaatatcaat atattaaatt agattttgca taaaaaacag actacataat actgtaaaac
     1081 acaacatatg cagtcactat gaaccaacta cttagatggt attagtgacc tgtagaatta
     1141 attcgagctc tagagctgca gggcggccgc gatatcccct atagtgagtc gtattacatg
     1201 gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca
     1261 caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca
     1321 aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg
     1381 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     1441 atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     1501 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     1561 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     1621 gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     1681 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     1741 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     1801 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     1861 aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc
     1921 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     1981 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     2041 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     2101 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg
     2161 taacactggc agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc
     2221 cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2281 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2341 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2401 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     2461 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2521 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2581 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2641 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2701 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2761 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2821 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2881 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgtt
//
