LOCUS       sequence-343170-        3792 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 3792)
  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 3792)
  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-kcnj10b
FEATURES             Location/Qualifiers
     source          1..3792
                     /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(1917..2016)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2033..2052)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2034..2052)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2057..2073)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2057..2073)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2175..2194)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2186..2995
                     /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(2253..2272)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3142..3730
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3631..3650
                     /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 accatgacct cggccacacc cccccctcat catggccccg cccctcagaa
      721 ggtctgccac tcccagacac agacagactt gctgaagcct ctgctcgggg gggcagggca
      781 tagcagtcat gtggcgcgca agaggcggat cctaaccaag gatgggcgga gcaacgtgcg
      841 cattgagcat gtgagtgggc tcagtgctct gtacctgcgc gacccctgga cgaccgttgt
      901 ggacatgcag tggagataca agctggttct gttcagcgcc acctttgtgg gcacctggtt
      961 caccttcggc ctgctctggt acctgctggc gctggtgcat ggagacctgc tcgagttcga
     1021 tcctccagca aaccacagtg tgtgtgtgat gcagatgcag actctgaccg gagcgttcct
     1081 cttctcgctg gagactcaga ccaccatcgg ttacggctat cgctgcatca cggaggaatg
     1141 tccgtccgcc atcgtcctgc tcatcataca gctgctgatc accaccgcca tggagatctt
     1201 catcaccggg acattcctgg ccaaagtggc tcgaccgaaa aagcgcggcg agacgattcg
     1261 cttcagtcag catgctgtaa tagccaatca cgacggccgg ccgagtctca tgatccgagt
     1321 ggcaaacatg cggaagagcc tcctgctggg ttgccaggtg acggggaagc tcttacagcc
     1381 gtgtgtgccg aaggcgggcg aaacggtgcg tctggaccag aaaaacgtgt cgttcagtgt
     1441 ggacacggcc agcgagagcc ccttcctcat cctgccgctc actttctacc acattattga
     1501 tgacgagagt ccactgcggg cctgggcgga caaaggcggt ggctggactg atcccggtca
     1561 gacagatttc gagctgctgg tgatgatgag ctctacagtg gagcccacct ctgctacctg
     1621 tcaggtgcgc acctcttacc tgcccgatga gatcctgtgg ggctacgagt tccctccaat
     1681 cgtctccctc tcgcctatgg ggaaatacgt ggcagatctg gcgtacttcg acaaagtggt
     1741 gaagaccaaa gccccgcccc ttttcgagca ggccccaccc actggcactg acatgggctg
     1801 cggggaggag agcacagacc cggagaagat gcagctggag cagagttacc gggacagaac
     1861 tggagacaaa ccggcggatg gcggctcgct caccgtgcgt atcagtaacg tctgagaccc
     1921 agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga
     1981 acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag
     2041 tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct
     2101 ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta
     2161 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg
     2221 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     2281 gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct
     2341 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     2401 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     2461 atggttactc accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc
     2521 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     2581 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     2641 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     2701 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
     2761 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     2821 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     2881 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     2941 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga
     3001 attggttaat tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag
     3061 ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt
     3121 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3181 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3241 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3301 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3361 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3421 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3481 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3541 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3601 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3661 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3721 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3781 tgctcacatg tt
//
