LOCUS       sequence-342866-        3873 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 3873)
  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 3873)
  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-kcnj2b
FEATURES             Location/Qualifiers
     source          1..3873
                     /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(1998..2097)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2114..2133)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2115..2133)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2138..2154)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2138..2154)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2256..2275)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2267..3076
                     /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(2334..2353)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3223..3811
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3712..3731
                     /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 accatgggaa gcgtgagagc ccaccgatac agcatagttt cctcagaaga
      721 aggtgggatg aagttgtccc ccattgctat tcaaaacggc tacggcaatg gaaaagtgag
      781 caccacagat cacgagcaac agcaaacgca aaaccgattc gtcaacaagg atggacgctg
      841 caatgtacag ttcatcaaca tgagcgaaaa aggtcagcga tacctagctg atatcttcac
      901 aacatgcgtt gacattcgct ggcgttggat gatggtcctt tttttcctgt ccttcctatt
      961 gtcttggctt atttttggac ttatattctg gctcgttgca tatacatctg gcgacttgga
     1021 taatcaagtg cagatatgca tttcgaatgt tgacagtttt actgcggctt tcctgttttc
     1081 ggtagaaaca caaacgacta ttggatatgg ctaccgctat gtgacggagg actgccccat
     1141 tgctgtgttc atggtggttt ttcagagcat cctgggatgc atcattgatg cattcatcat
     1201 tggtgcggtc atggctaaga tggccaagcc taaaaaacga aatgaaaccc tagtgttcag
     1261 tcaatatgca acaatcgcaa tgagggacgg taaactgtgc ctaatgtgga gagtcgggaa
     1321 cctccgcaaa agtcacctgg tggaggcaca cgtccgcgct cagctgctca aatcccgcac
     1381 cacagctgag ggcgaattca tccctctgga tcagttagac attgatgtcg gattcgacac
     1441 tggtattgac cgtatatttt tagtgtcccc catcaccatc gtccatgaga ttgacgagga
     1501 cagtccattc tatgacatga gcaaacagga actggaaagt tccgagctgg aaatagtggt
     1561 gattttggag ggcatggtgg aagctacagc catgaccact cagtgtcgaa gctcttacgt
     1621 gactagcgaa atcctgtggg gccatcgctt tgagccggtc ctttttgagg agaagaatca
     1681 ttacaaggtg gactactcgc acttcgaaaa cacctacgag gttcccaaca cacctcaatg
     1741 cagcgctcga gatttggcag agaagacgta cataatgtct aattcaaact ctttttgtta
     1801 cgagaatgaa gtcgctttca tggataaaga ggaaacggag gatgatgatg atgacgatga
     1861 tgatgagcag attaacatca ggagggacag tgtggtttta gagggatcca atatggacga
     1921 tgtcaaaagc acagcttcta gtcaggacac tttaccacta cagctcaaac ctttaagaca
     1981 acagcctgag atctgagacc cagctttctt gtacaaagtt ggcattataa gaaagcattg
     2041 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     2101 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     2161 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     2221 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     2281 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     2341 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     2401 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     2461 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     2521 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     2581 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     2641 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     2701 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     2761 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     2821 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     2881 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     2941 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     3001 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     3061 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     3121 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     3181 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3241 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3301 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3361 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3421 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3481 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3541 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3601 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3661 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3721 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3781 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3841 cctggccttt tgctggcctt ttgctcacat gtt
//
