LOCUS       sequence-343327-        3735 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 3735)
  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 3735)
  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-kcnj20
FEATURES             Location/Qualifiers
     source          1..3735
                     /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(1860..1959)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1976..1995)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1977..1995)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2000..2016)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2000..2016)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2118..2137)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2129..2938
                     /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(2196..2215)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3085..3673
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3574..3593
                     /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 accatgcaga gtgaggcctc caacccctat gtttccatac gccgccacaa
      721 gagcaccaca gatctgccat acagcccaca gcgtgggtac ttgcgtggac agagaactag
      781 tatctgcgcc cccaccgagg ccgagctggc aatgatgagg cgttgctcct tatgtgatcc
      841 gaacgttcgt ccatgttgcc gccatgctct tagcgttccc aatatgacca gtccaagcgc
      901 tagccctctg tacagcaccc catcccgcag tgctcactgc actcctgtac gggaagtgga
      961 gtgcaatgct aaagcccgca ataagctaat tgaaaacgag agcgtgcaaa ctcccacatg
     1021 cccaccagag attcgccagc agcgtcgcta cgtctcaaaa gatgggaaat gccgagtaga
     1081 cttggcacac atggcagaaa aggagcgatt tctggcggac atcttcactt cgtttgtgga
     1141 cctccagtat agatggtttc tgttcgtatt catgatgtgc tacacggtca cttggtttat
     1201 attcgcaggg ctttattcgc taaatgcttt cctgcgtgga gatcttgaac atcctgcgga
     1261 tactgatgtt tgcttccaaa atatagatgg atttgtttca gcgttgctct tttcagtgga
     1321 aacacagagg actattggtt atggcgctcg tacagtctca ccacgctgct atgaaggtgt
     1381 gtttttggtc atggcgcaat gcattgtggg atccatgatt gacgctctaa tggtgggctg
     1441 catgtttgtt aaaatctcac gtcctaagaa gcgagccgaa acacttcttt ttagtcggac
     1501 ttgtgttgtg gccaatcggg atgaccgtct ctgccttatg tttcgtcttg gagacttgcg
     1561 tgagagtcac atggtagacg ccaaagtacg tgctaaactc atcaagtctc ggcagacatc
     1621 ggaaggggaa tttctccccc tggagcaatc ggagatcgac ctgggctatg agagcggctc
     1681 tgaccgactg tttctagtgg agccgcaagt gattcaacac atcattgatt ccagcagtcc
     1741 tttctgggag ctcgggccgg agaagctgcg gaagcaacag tttgagatta ttgtcatctt
     1801 ggagggcata gtggaagcta caggtggcat tatctgcatg tgtctgttga cccagtaaga
     1861 cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa
     1921 cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg atatccccta
     1981 tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa
     2041 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     2101 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc
     2161 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     2221 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt
     2281 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     2341 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     2401 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata
     2461 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     2521 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     2581 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     2641 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt
     2701 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     2761 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     2821 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     2881 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     2941 agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc
     3001 aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     3061 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3121 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3181 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     3241 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3301 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3361 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3421 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3481 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3541 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3601 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3661 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3721 ttttgctcac atgtt
//
