LOCUS       sequence-343157-        3801 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 3801)
  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 3801)
  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-kcnj8
FEATURES             Location/Qualifiers
     source          1..3801
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     12..29
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(188..215)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(307..393)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     442..464
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     456..473
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     457..473
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    489..588
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1846..1945)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1962..1981)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1963..1981)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1986..2002)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1986..2002)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2104..2123)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2115..2924
                     /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(2182..2201)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3071..3659
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3560..3579
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
       61 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
      121 aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa gagtttgtag
      181 aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta
      241 tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca aatccgctcc
      301 cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc
      361 ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcgt
      421 taacgctagc atggatgttt tcccagtcac gacgttgtaa aacgacggcc agtcttaagc
      481 tcgggcccca aataatgatt ttattttgac tgatagtgac ctgttcgttg caacaaattg
      541 atgagcaatg cttttttata atgccaactt tgtacaaaaa agcaggcttc accatgttgc
      601 cgcggaagag catcatcccg gaggagttcg cgctgccggc gctggtgtcc cgggtgccgc
      661 gtaaaccagt gttcagagac cgcgtgaata aagcgcgctt catcgccaag agcggcgcgt
      721 gtaacttggc gcacaagaac atccgtgagc agggccggtt cctgcaggac gtcttcacca
      781 ctctggtgga tctgaagtgg cgcttcacgc tggtcatctt caccatgaca tttctgtgca
      841 gctggctgct gttcgccatg agctggtggc tggtggcctt cgggcacgga gatctggaca
      901 ccgaccgtaa acctgccgtg gagcagtgcg tcaccaatgt caactctttc acctctgcct
      961 tcctgttctc catcgaggtg caggtgacta taggtttcgg agggcgtatg ataacagagc
     1021 ggtgtcccat cgccatcgcc tttctcatat tgcagaacat cattggcttg ataataaatg
     1081 ccgtcatgct tggctgcatc tttatgaaga ccgctcaatc ccaccggcgt gccgaaaccc
     1141 ttatctttag ccgacacgca gtcatcgctg tgcgcaacaa ccgcctctgc tttatgatcc
     1201 gagtgggcga cctgcgcaaa agcatgatca tcaatgcagt tgtgcgcctc caagtggtcc
     1261 ggaagaccat gaccccagag ggcgaggtca tccccatcca gcagatcgat gtccagactg
     1321 agagtgcagt ggctggtaac agcatctttc ttctggcacc cttgatcata tgtcatgtca
     1381 ttgataaaga cagtcctctg tacgatctgt cggcgatgga gctgcagtgc agtgatctgg
     1441 aggtcatcgt gatcctggaa ggagtggtgg agaccacagg catcaccact caggctcgca
     1501 cctcctacgt gaccgaggag atccagtggg gacacagatt cgtgcctata gtgacagagg
     1561 aagagggtgt ctactcagtg gactactcca agttcgggaa tacagtcaaa gtggccaccc
     1621 ctcgctgcag cgctcgtgag ctggacgaga aaccctccat cctgatccag acgctgcaga
     1681 agagcgagct ttcacaccag aactctctgc gcaagaggaa ctctatgcgc aggaacaact
     1741 ccatgcgcaa gggcaactcc atgcggcgca ataattcagc tctcgctgtg cccaaagtgt
     1801 agttcctcac ccctgagggc ggagccaatc tagcggtcac atgagaccca gctttcttgt
     1861 acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta
     1921 tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt
     1981 acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca
     2041 ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta
     2101 atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca
     2161 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
     2221 cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
     2281 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
     2341 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     2401 ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg
     2461 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
     2521 attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata
     2581 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     2641 tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     2701 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     2761 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     2821 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     2881 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt
     2941 ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa
     3001 aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca
     3061 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3121 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3181 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     3241 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3301 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3361 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3421 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3481 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3541 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3601 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3661 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3721 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     3781 ataccgctcg ccgcagccga a
//
