LOCUS       sequence-343298-        3867 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 3867)
  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 3867)
  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-kcnj12a
FEATURES             Location/Qualifiers
     source          1..3867
                     /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(1992..2091)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2108..2127)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2109..2127)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2132..2148)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2132..2148)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2250..2269)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2261..3070
                     /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(2328..2347)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3217..3805
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3706..3725
                     /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 accatgagcg tgggtcggat tggtcgctac agtatagtgt cgacggagga
      721 ggatggtctg cgtttgacga ccatgcatgg tggagggatg aatggatacg gcaatggcaa
      781 aatccacacc cggagaaaat cccgcaaccg cttcgtcaag aagaacggac agtgcaatgt
      841 gcagttcacc aacatggacg agaagtccca gcgctacatg gcggacatat tcaccacttg
      901 cgtggacata cgttggcgct acatgctaat cgttttcacg cttgttttcg tgatttcctg
      961 gttagcgttt ggcttggcgt tttgggttat cgcgcttctt cacggtgacc tggacaaccc
     1021 agcaggtgat gacaacttta cgccatgcgt gctacaagtc aacggcttca tcgcagcgtt
     1081 cctgttctct atcgagacgc agacaacaat cggctacggt ttccgctgtg taacagaaga
     1141 atgtcctctc gctgtgtttc tcgtggtctt ccagtccatc gtgggcagca tcatcgactg
     1201 cttcatgatt ggagcaatca tggccaaaat ggcacgtccc aagaagcggg cacaaactct
     1261 gctgttctcg cataacgccg tcatcgccat gcgtgatgga aaactgtgct tgatgtggcg
     1321 ggtcggcaat ttacgaaaga gtcacattgt tgaggcgcac gttcgggcac agctcatcaa
     1381 accgcgggtt acgactgaag gagagtacat cccgctcgac caactggaca taaacgttgg
     1441 tttcgataaa ggacttgacc gcatcttcct cgtctctcca atcaccatcc tgcatcaaat
     1501 tgaccacgag agtcctctgt tcggcattag taagcaggat ctggaaacgg cagattttga
     1561 gatcgtggtc atcttggagg gcatggtgga ggcgacagcc atgacggcac aggctcggag
     1621 ctcctacttg gccagtgaga tcctctgggg gcatcgcttt gagccggtgc tgtttgagga
     1681 aaagaaccag tacaaggttg attactctca cttccacaag acctacgagg tgccgtccac
     1741 ccctcgctgc agtgccaagg acatgatgga aaacaagtac ctggtggttc ccaccgctaa
     1801 ctccttctgc tatgagaacg agctggcgat cctgagccac gaagaagaag aggacgacag
     1861 tccagaacgg ataaagccag agcgagcaat gagcctcagc ccggagagaa cccctcgaca
     1921 tgacttcgac cggctgcaga cgcctcgatg cacagaacaa aggtcatacc gcagggagtc
     1981 ggagatatga gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     2041 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     2101 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     2161 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     2221 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     2281 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     2341 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     2401 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     2461 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     2521 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     2581 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     2641 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     2701 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     2761 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     2821 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     2881 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     2941 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     3001 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     3061 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     3121 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     3181 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3241 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3301 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3361 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3421 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3481 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3541 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3601 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3661 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3721 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3781 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3841 cttttgctgg ccttttgctc acatgtt
//
