LOCUS       sequence-343299-        4182 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 4182)
  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 4182)
  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-kcnj14
FEATURES             Location/Qualifiers
     source          1..4182
                     /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(2307..2406)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2423..2442)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2424..2442)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2447..2463)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2447..2463)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2565..2584)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2576..3385
                     /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(2643..2662)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3532..4120
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4021..4040
                     /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 accatggggg ccgctcgagt gaagcggcgc ttcagtgcgg ttgtggacac
      721 ccctctggat gaggaagaag tcatgaggct ggcacaaagt gatgatggca cttcctccca
      781 ttcgggcaca cttccatctc ccaactgcaa tggcaaagtg ctcagcattc tccacagcaa
      841 tggaggaagg agcggagaga gagatgagga tgagaatgat gatgatgatg cggacaagga
      901 tgatgatgat gaggaggaag ggggaggaag gaggggaggg ggaggacgtg gctcggcact
      961 cagtggggag tctggggttg aagggaggtg ggaaaggaac aaaagagcat ccttaccttc
     1021 cctttccccc acaccgctcc gtgatcgccg ccgccagtcc cgccggcccc gaaggcgctt
     1081 tgtaggcaag gacgggcgct gcaacgtcac cttcgtcaac atgagcgaaa ggggccagcg
     1141 ctacctaact gacctcttca ccacctgcgt tgacatccgc tggcgctgga tgctcgtggt
     1201 gttcaccctc tccttcttgc tcacctggct tctctttggt ttcacgttct ggctcattgc
     1261 tgctgctcat ggggatttga atcccttagc atcatcttcc tcttctccct catctctttc
     1321 atcctctctg gctccactgg atgccaccga aggttccagt cgagtggaaa tggtagcaga
     1381 gacagaggaa ccctgctttc agcaggtgaa cagcttcatg gcggcgttcc tcttctctct
     1441 agagacacag acatctatag gttatggctt ccggagtgtt accgaggcct gccccctggc
     1501 ggtgttggct gtcgttttgc agtgcattgt gggctgcatc atcgacgctt tcatcattgg
     1561 ggcggtaatg gctaagatcg ccaagcccaa gaagcgcaat gagacactgg tgttctcgga
     1621 tgtggctgtg gtggccgtga gggacgggaa actctgcatg atgtggcgag tgggaaactt
     1681 gcgtaagagc cacctggtgg aggctcatgt tcgagcacag ctacttaagc cacgggtgac
     1741 ccctgagggc gagttcctgc cattggacaa ctcagacctc aatgtcggct ttgacacggg
     1801 aacagatcgc atctttttgg tgtcacctgt gaccattgta catgaaatta atgaggagag
     1861 ccccttcttt gaaatggaca aaggtgcact agagggagac aaagatttgg aagtggtggt
     1921 aattctggaa ggtatggtgg aggccactgc catgaccact caatgtcggt cctcttttct
     1981 ggcttctgag attctctggg gtcaccgatt tgagcctgtg ctctttgaaa gaaagaactg
     2041 ctaccaggtg gactactcat acttcaacag gacttatgag gtcacggaca ctcccacctg
     2101 cagcgctaag gagttagcag agaaaaagta cattgcaggc tcccgttcct ccttctgcta
     2161 cgagaatgag gtggctctgc agctctcctc ctctgtcccg ccctgcgctg accagttctc
     2221 tccatccccc tgttcgtccc cccctcagtc acccacacca ccacctcgac aagactcctg
     2281 cagtgaacac ccacacaaac actgagaccc agctttcttg tacaaagttg gcattataag
     2341 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     2401 tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc
     2461 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     2521 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     2581 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata
     2641 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta
     2701 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     2761 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     2821 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa
     2881 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     2941 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     3001 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     3061 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     3121 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     3181 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     3241 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     3301 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     3361 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag
     3421 cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta
     3481 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3541 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3601 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3661 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3721 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3781 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3841 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3901 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3961 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4021 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4081 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4141 tttacggttc ctggcctttt gctggccttt tgctcacatg tt
//
