LOCUS       sequence-343310-        3816 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 3816)
  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 3816)
  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-kcnj12b
FEATURES             Location/Qualifiers
     source          1..3816
                     /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(1941..2040)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2057..2076)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2058..2076)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2081..2097)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2081..2097)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2199..2218)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2210..3019
                     /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(2277..2296)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3166..3754
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3655..3674
                     /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 tgggcaagcc ccaccgctat aacatcgtgt catcgtcgga
      721 ggaggacgtg ggccgtcacg gcaccatgcc cgccctcggc aatggcttcg gaaatggcaa
      781 gatccagacc cggcggaagc tgcggagccg cttcgttaac aaaaccggcc aatgcaatgt
      841 cagcttcgct cacatggacg agcaatccca gcgatacctc gctgacatct tcaccacctg
      901 cgtggacagt cgctggcgct ggatgtttct acttttctcc ctcgcatttg tcatctcctg
      961 gttagcgttc ggatttgcgt tctggcttat cgctcttgtc catggtgatc tagagaaacc
     1021 tcaagaaaac ttcacgccat gcgtcatgca ggtcaacagc ttcgtcgcag cgtttttgtt
     1081 ctcagttgag acacaaacaa caattggcta cggatttcgc tgcgtgactg aggagtgtcc
     1141 tttagctgtg tttatggtcg tcttccagtc cattgtgggc tgcattattg actcgttcat
     1201 gattggtgca atcatggcca aaatggcgcg acctaaaaag cgtgcagaaa cgatactgtt
     1261 ctcgcataac gccgtcatct caatgcgcga tgggaagctc tgcctcatgt ggcgggttgg
     1321 gaacttgagg aagagtcaca ttgttgaggc tcatgtgcgg gcgcaactta ttaaaccccg
     1381 aattacagaa gagggcgagt acataccttt ggatcaaatt gacatcaatg ttgggtacga
     1441 ccaaggtctg gaccgcatct tcttggtcgc tcccctcacc atcctccatg agataaatga
     1501 ggagagtccg ctgtttggaa tcaaccagca ggatctggaa acatctgatt ttgagattgt
     1561 ggtcatactg gaaggaatgg tggaggcgac tgccatgacg gctcaggtgc gcagctccta
     1621 cctgtccagt gagatcttat ggggccatcg ctttgagcct gtggtattcg aggagcgcaa
     1681 ccagtataag gtggactatt cgcactttca taaaacctac gaggtgccct cgacgccacg
     1741 ctgcagcgcc aaagacctgg aagacaacaa atccttggag tccggtgcca acttctgcta
     1801 tgagaatgag ttagctttta ttagcaggga tgaagaggag caagaggagg acaggggtga
     1861 gagggggact gagctagaga cactttcagc caatctgaac tttgatcaga ggtcgtatca
     1921 caaagagtct gaaatttgag acccagcttt cttgtacaaa gttggcatta taagaaagca
     1981 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     2041 ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
     2101 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     2161 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     2221 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     2281 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     2341 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     2401 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     2461 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
     2521 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     2581 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     2641 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     2701 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     2761 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     2821 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     2881 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     2941 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     3001 gctcgatgag tttttctaat cagaattggt taattggttg taacactggc agagcattac
     3061 gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc
     3121 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3181 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3241 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3301 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3361 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3421 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3481 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3541 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3601 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     3661 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     3721 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     3781 gttcctggcc ttttgctggc cttttgctca catgtt
//
