LOCUS       sequence-343332-        3850 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 3850)
  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 3850)
  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-kcnj21
FEATURES             Location/Qualifiers
     source          1..3850
                     /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(1975..2074)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2091..2110)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2092..2110)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2115..2131)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2115..2131)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2233..2252)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2244..3053
                     /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(2311..2330)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3200..3788
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3689..3708
                     /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 accatgagtg gtttcaaggg ccaggatgtg gaaaaacagg gcatcaagtt
      721 gccaaaaaaa atggattgat cacaatgaca aagtccccac agaccgtaca atggtggttg
      781 ggggagtcgt gggtggagga gattgttctg aacccgtagg agctcctctg aaaaaacgcc
      841 gccagcgata cgtggagaaa gacggcaagt gcaacgtcca ccacggaaat gtgcgtgaaa
      901 cctaccgcta cctgacagac atcttcacca cgctagtgga tttaaagtgg cgcttcaatc
      961 tgctggtctt cacactggtc tacaccacca catgggtgtt ttttggcttc atctggtggc
     1021 tcatcgccta cgcccgagga gatctggagc acactgatga taatgactgg acaccttgtg
     1081 tgaataacct caatggcttc gtctcagcgt ttcttttttc catagagact gagaccacca
     1141 ttggttacgg gtatcgtgtg ataacagaca aatgtccaga ggggatcatc ctgcttctga
     1201 ttcaggccat tttgggctcc atcgtcaacg cctttatggt gggctgcatg tttgtgaaaa
     1261 tctcccaacc aaagaaacga gccgaaacac tcatgttttc caacaccgct gtgatatccg
     1321 tgcgggacag caagttgtgt ttgatgttcc gtgtcggtga cttgcgtaac tcgcacattg
     1381 tggaggcctc cattagggcg aagcttatcc gctcgaaaca gacgaaggaa ggagagttta
     1441 ttcctctcaa ccagacggac atcaatgtgg gctttgacac gggggacgat cggctgttcc
     1501 tcgtgtcgcc gttgattatt tgccacgaga tcaacaagaa cagcccattc tgggacgtct
     1561 ctcaggagca tcttgtacga gaggagtttg agatagtggt catccttgaa ggaatggtgg
     1621 aagctacagg aatgacgtgt caggcgcgca gctcctacct gagctcagag gtgctgtggg
     1681 gcgagcgctt cacccctgtg ctctcactgg aggagggttt ctacgaggtg gattatgaca
     1741 ccttccacca cacctatccc gccccgacac catcctgctc cgccagagag ctcgcagaac
     1801 tgctcaagaa aggggaatcc atacctctgc cccctatttc acctccagcg gccctagcgg
     1861 agccagcaag ccctgttgag acgccggagg agaaggagga gaatatagat gaagacgcag
     1921 aagagactgt tagcaatggg gatgtgaaca gaatggagat cgagcatgaa tgagacccag
     1981 ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac
     2041 aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg
     2101 agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct
     2161 gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca
     2221 taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat
     2281 taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc
     2341 aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga
     2401 aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc
     2461 tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat
     2521 ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg
     2581 attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc
     2641 ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac
     2701 gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg
     2761 ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca
     2821 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta
     2881 ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact
     2941 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata
     3001 atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat
     3061 tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct
     3121 catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag
     3181 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     3241 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     3301 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     3361 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     3421 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     3481 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     3541 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     3601 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3661 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     3721 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3781 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     3841 ctcacatgtt
//
