LOCUS       sequence-343178-        3696 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 3696)
  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 3696)
  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-kcnj11
FEATURES             Location/Qualifiers
     source          1..3696
                     /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(1821..1920)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1937..1956)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1938..1956)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1961..1977)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1961..1977)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2079..2098)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2090..2899
                     /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(2157..2176)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3046..3634
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3535..3554
                     /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 accatgttgt ccagaaaagg actcattccc gaggattatt tgttgactcg
      721 gttggcggag gatgtgttac agccaaaatt caagactaaa cagcggaaag cgcgattcgt
      781 cgccaaaaac ggaacgtgca acgtcgcgca cacgaacatt cgcgaacagg gacggtttct
      841 acaggacgtg ttcaccactt tggtggatct gaaatggctt catactctgg tgatcttcac
      901 catgtccttc ctgtgcagtt ggcttttgtt cgggatgatc tggtggctga tcgccttcgc
      961 gcacggagac ctggatttga aaggcgagga ctttgtgccg tgcgtaacgg acatccactc
     1021 gttctcctcc gcctttctgt tctccatcga ggtgcaggtg accatcggct tcggaggacg
     1081 gatgatcacg gaggaatgcg tttcggccat cgtcatactt attcttcaga acatagtggg
     1141 tcttgtgata aacgccatca tgttaggatg catcttcatg aaaacggcgc aagcaaaccg
     1201 gcgagcggaa acgctcatct tcagcaaaca cgcggtgata actgtccgaa acaacaagct
     1261 gtgcttcatg tttcgccttg gtgacctgag gaagagcatg attatcagtg ccaccgtcca
     1321 catgcaggtg gtacgaaaga ccgtcaccag cgaaggcgag atggtgcctt tggaccagat
     1381 agacatccag atggacaacc cggtgggcac caacagcatc tttctggtgt cccccctcat
     1441 catcagtcac accattgaca aaaacagccc tctgtatgat ctgtccgcgg acgatttact
     1501 gcatgaggac atcgaggtga ttgtcgtcct ggaaggggtc gtggagacca ccggcatcac
     1561 cactcaagcg aggacctctt acctgtccga ggagatttta tggggtcata ggtttgtgcc
     1621 tacggtgtct gaggaggacg ggatgtatgc tgtggactac tcgaagtttg gaaacacggt
     1681 taaagtggcg acgccgaaca gcagtgcgcg gactctcgag gaagcgggag ggatggagag
     1741 gttcaaactg caggagacca gcggcgcatg cgcagcagtg aggagaagag tcaaacaccc
     1801 aaacaccaaa gaaatgtgag acccagcttt cttgtacaaa gttggcatta taagaaagca
     1861 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     1921 ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
     1981 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     2041 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     2101 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     2161 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     2221 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     2281 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     2341 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
     2401 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     2461 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     2521 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     2581 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     2641 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     2701 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     2761 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     2821 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     2881 gctcgatgag tttttctaat cagaattggt taattggttg taacactggc agagcattac
     2941 gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc
     3001 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3061 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3121 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3181 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3241 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3301 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3361 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3421 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3481 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     3541 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     3601 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     3661 gttcctggcc ttttgctggc cttttgctca catgtt
//
