LOCUS       sequence-343129-        3828 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This gene was cloned into the gateway middle entry vector. It can be
            used for generating antisense riboprobe or gene expression using a 
            gateway system..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3828)
  AUTHORS   Silic MR, Murata SH, Park SJ, Zhang G
  TITLE     Evolution of inwardly rectifying potassium channels and their gene 
            expression in zebrafish embryos.
  JOURNAL   Dev Dyn. 2021 Sep 23. doi: 10.1002/dvdy.425.
  PUBMED    34558132
REFERENCE   2  (bases 1 to 3828)
  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: "Dev Dyn. 
            2021 Sep 23. doi: 10.1002/dvdy.425."
COMMENT     Sequence Label: pDONR221-kcnj5
FEATURES             Location/Qualifiers
     source          1..3828
                     /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    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1953..2052)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2069..2088)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2070..2088)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2093..2109)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2093..2109)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2211..2230)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2222..3031
                     /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(2289..2308)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3178..3766
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3667..3686
                     /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 tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggcttc accatggcag gagattctcg cgtccacatg gaccacgaca tggagatagg
      721 agtgactcca agagagcagg tgaagaaact ccccaaacac ctgagggaat cccagatccc
      781 cacagaccgt acccacctca tctctgatcc ggttaagaaa ccccgtcagc gctacgtcca
      841 gaaagacggc aagtgcaacg ttcatcacgg aaacgttaaa gagacctacc gttatctcag
      901 cgacctcttc acgacactag tggacttacg atggcgcctc agtttcttca tcttcacctt
      961 ggtctacgtg gtcaactggc tgttttttgg gtttttatgg tggctaattg cactgatacg
     1021 aggagatctt ctacacagcg acgaggaggg ctggaccccc tgcgttgaaa acctcaacag
     1081 ctttgtatcc gctttcctgt tttccatcga aaccgagacc actattgggt atggctatcg
     1141 ggtgatcacg gagaagtgtc cggaagggat aatacttcta atggtgcaag ccatattggg
     1201 gtcgattgtc aacgccatga tggtagggtg tatgtttgta aagatttcac aacccaagaa
     1261 ccgagcagag acgttgatgt tctcccacaa tgctgtgatc tcggtgcggg acaacaagat
     1321 gtgcctgatg ttcagggtgg gcgacttgag gaattcccac attgtggaag cgtcaattcg
     1381 ggccaaactc atccgctcgc agcagaccaa ggagggcgag tttattccgc tcaatcagac
     1441 agacatcaac attggcttcg acacaggaga cgatcgctta ttcttggtat ctcctcttat
     1501 catctcacat gagatcaacg agaagagtcc tttctgggag atgtcgcggg cacagatgga
     1561 gaaggaagag ttcgagattg tggtcatctt agaaggcatg gttgaagcta caggcatgac
     1621 gtgtcaggcg cgcagctcgt acttggactc ggaggtgctg tggggcttcc gcttcacgcc
     1681 ggtgctttcg ctggaaaagg gcttctacga agtcgactac aacaactttc acgacatata
     1741 cgaaaccaac accccgacct gtagcgctaa agaactctcc gcccggttaa aagagagccc
     1801 cctgctcccg cacctgcccc tgctcagccc ggacctcagg gcgcacacaa cggacccgtt
     1861 ccagaccacc gaggaccagg ccaagcacga cgccgacacc ccgccggaga ccaacaacgg
     1921 ctcggcaaac cgcctggagg agaatccctg agacccagct ttcttgtaca aagttggcat
     1981 tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa
     2041 aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc
     2101 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     2161 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     2221 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     2281 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     2341 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     2401 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     2461 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     2521 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     2581 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     2641 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     2701 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     2761 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     2821 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     2881 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     2941 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     3001 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     3061 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     3121 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3181 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3241 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3301 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     3361 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3421 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3481 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3541 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3601 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3661 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3721 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3781 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtt
//
