LOCUS       sequence-342841-        3708 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 3708)
  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 3708)
  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: pENTR-D-kcnj1b
FEATURES             Location/Qualifiers
     source          1..3708
                     /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(1833..1932)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1949..1968)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1950..1968)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1973..1989)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1973..1989)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2091..2110)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2102..2911
                     /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(2169..2188)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3058..3646
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3547..3566
                     /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 agcaggctcc gcggccgccc ccttcaccat gttccagttc atccggaaac agcttcacaa
      721 tcacctggtg gagcgcaaaa tcaggagaac tcgcctcgtc accaaagacg gccactgcaa
      781 catcaaattc ggcaccaccg aatatcaaag ccacttcgct ttccttaagg acttctggac
      841 tacattcgtc gaaatccact ggtgtttcgt aatcctttta tttgccgcag ccttcaccgg
      901 cagctggttc gttttcggcc tgctctggta ttcgctagcg aaaagcaacg gagacctgtc
      961 tgagtctcag cagacgaaat gcatcgagaa cattaacagc ctcacgactg ctttcttgta
     1021 ttctttagaa acccaaacca ctattggata tggcggtcgg gctcttacgg ggcactgtcc
     1081 tggcaccgtc gctctattaa taatccaatc cctcatgggc gccatcatca actgcttcat
     1141 gtgtggactc atcctggcca agatctcgct cccgaagaga agagcaaaga ctgtagcgtt
     1201 cagcgaaaca gccgtcatcc gtctctggaa ggggaattta tgcctgcaga tacgagttgc
     1261 caatctacgg aaaaccctac tgattggaag ccacatttac ggaaagcttc tgcggactac
     1321 tatcactccc gaaggtgaaa ctatcattct ggatcaagta aatgttgact tttcggtgga
     1381 cgccgggaag gataacttgt tttttgtgtg tccgcttact atttatcatg tgatcgacaa
     1441 atccagtccg ttcgcggaca tggcagcgga taaccttcag ctccaggact ttgagttggt
     1501 ggtttttctt gacggcacgg ctgaatctac aagcttgtcc tgccaagtgc gaacttctta
     1561 ccttcccaga gagatcaaat ggggttacgt gtttctcccc gtcatctctc gcaccaaaac
     1621 tggaaaatac cgtatcgatt tatccaattt ttccaaaacg cagcaagcag atactccaca
     1681 ttgtgcttgc tgttttcaca acaaacataa caaggggctc attctccacc ggcatggaag
     1741 cgaaggcatc gataacctgg gatttgacgt tattgacctt caggactcgg ctgatgtaac
     1801 tgagaagggc gattaaaagg gtgggcgcgc cgacccagct ttcttgtaca aagttggcat
     1861 tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa
     1921 aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc
     1981 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     2041 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     2101 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     2161 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     2221 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     2281 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     2341 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     2401 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     2461 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     2521 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     2581 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     2641 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     2701 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     2761 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     2821 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     2881 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     2941 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     3001 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3061 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3121 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3181 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     3241 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3301 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3361 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3421 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3481 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3541 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3601 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3661 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtt
//
