LOCUS       sequence-343128-        4047 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 4047)
  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 4047)
  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-kcnj4
FEATURES             Location/Qualifiers
     source          1..4047
                     /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(2172..2271)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2288..2307)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2289..2307)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2312..2328)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2312..2328)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2430..2449)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2441..3250
                     /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(2508..2527)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3397..3985
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3886..3905
                     /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 accatgggga ccaaccgagc caacaggtac agcatcgtgt ctccagatga
      721 ggagcggctg aagatcgcca ccctcggcct ccacaacggc cacggttccg cccgcctaca
      781 ctcgccggtc tccccccgcc tgctgtctcc gagccccggg gcctctagca ctgcagtcac
      841 cggctccggc ttcaatggga agatctccac gtccggccgg agccagctcc gcagccgctt
      901 cgtcaagaag aacggccagt gcaacgtggt gttcgccaac atggaggaca agccgcggcg
      961 ctacctggca gacttgttca ccacctgcgt ggacatccgc tggcgctacc tgatgctggt
     1021 cttctgctcc acgttcctgc tgtcctggct cctgttcggc gtcatcttct acagcgtctc
     1081 tctgcttcac ggagacttcc agaaagagca cggaaagctc tggaaaccgt gtttgcttca
     1141 tgttgaaggg ttcgtcgggg cgtttctgtt ctccatcgag actcagacta cgattggcta
     1201 cggctggcgc tgcgtgactg aggagtgtcc tgtagcgata gtgaccgtag tggtgcaatc
     1261 gatcgtcgga tgcattattg actccttcat gattggaacc atcatggcta aaatggcacg
     1321 tccaaagaag aggaaccaaa ccttgatgtt ttcccaaaat gccgtgatcg cgttacgcga
     1381 tgggaagctg tgtttgatgt ggcgcgtggg gaacctccgg cggagccaca ttgtagaggc
     1441 acatgtgcga gcgcagctca tccggccgta cgtaacagca gagggtgagt tcatccctct
     1501 ggagcagacg gatctgaacg taggctacga tgacgggacg gaccgcctgt tcctggtgtc
     1561 cccacttgtg atcgtgcacg agattgatga ggactctccc ctgtggggaa tgagccgagc
     1621 agaactagag tcagatgatt tcgagatcgt ggtgatcctg gagggaatgg tggaggcgac
     1681 agctatgacc acacaggcac gcagctccta cctgtcccgc gagatcctgt gggggcatcg
     1741 cttcgagcct gtgatcttcg aggaccacga ccgctatcag gtcgactacg cacattttca
     1801 taaaacctac gaggttccct cgacacccgg ctgcagcgct aaagagctcc acgagctgac
     1861 ccgccggcgg tcctcgaact cgtctcgctc tgggacgccg cagacgctga tgccgccgca
     1921 ctcgcccagc gccttctgct acgagaatga agtggctctg tgctgcgggg aggaggacga
     1981 ggagccgccg gaggaccaaa tgaacaccat tccctcagac ttccagaaaa tgctccagga
     2041 ctcggccgcc gtgagctccg gcagtaacat gctctgcatg ttagacatgg acaatcagat
     2101 cgagttcgac atcctccact ccaccatacc actcgacccc atgacatata agagcgaatc
     2161 cgagatatga gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     2221 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     2281 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     2341 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     2401 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     2461 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     2521 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     2581 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     2641 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     2701 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     2761 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     2821 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     2881 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     2941 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     3001 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     3061 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     3121 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     3181 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     3241 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     3301 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     3361 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3421 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3481 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3541 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3601 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3661 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3721 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3781 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3841 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3901 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3961 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4021 cttttgctgg ccttttgctc acatgtt
//
