LOCUS       sequence-343325-        4068 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 4068)
  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 4068)
  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-kcnj19b
FEATURES             Location/Qualifiers
     source          1..4068
                     /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"
     CDS             1772..1783
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(2193..2292)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2309..2328)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2310..2328)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2333..2349)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2333..2349)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2451..2470)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2462..3271
                     /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(2529..2548)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3418..4006
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3907..3926
                     /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 accatgtccg cgatccgcag aaaatttggc gaagactatc aggtggtgaa
      721 caccaaccgg gccatgtcct tttctgcccc tgctaaaagg aagcgtcagc ggtttgtgga
      781 gaaaaacggg cgctgcaatg tccagcacgg taaccttggt ggggagacca gcaggtacat
      841 ctctgacctt ttcaccacgc tcgtcgacct taagtggcgc tggaacctgc taattttcct
      901 tttaacgtac acggtcgcgt ggctcatcat ggcatcgatg tggtggatca tcgcgtacat
      961 ccgcggagac ctcggccacg gccacgacga ctcgtacacg ccgtgcgtcg ccaacgttta
     1021 caactttcca tcagcttttc tcttcttcat tgagactgag gcgactatag gctacgggta
     1081 ccgatatata acagagaagt gcccggaggg catcatactt ttcctttttc agtcgctgtt
     1141 gggctccatc gtggacgcgt ttctcatcgg ctgtatgttt attaagatgt ctcagcccaa
     1201 gaaacgcgca gagaccctca tgttcagcca ggacgcagtc atctcacaac gggacgggaa
     1261 gctctgcctc atgtttagag tcggaaatct caggaatagc cacatggtgt cggcgcaaat
     1321 caggtgcaag ctcatcaagt ctcgtcaaac cccagaggga gagtttctac cattggatca
     1381 gcgggaattg gatgttggat ttggaactgg tgctgaccaa cttttcctcg tctcacccct
     1441 cacaatatgt cacgagatca attccaatag cccctttttt gacctgtccc agcgctccct
     1501 tatgaacgaa cagtttgaga tcgtcgtcat cctggaagga atcgtggaaa ctacaggcat
     1561 gacgtgtcaa gctcgaacct cttatacaga ggatgaagtg ctgtggggtc accgctttct
     1621 accagtcatg tcgctagagg aaggattctt cagagtcgac tactcacagt tccacaacac
     1681 atttgaggtg cccactcctc catacagtgt gaaggaacaa gatgaaaagt cctccctgcc
     1741 ctcccctggt cctgttctct cccctacatt gattgaaggt cgtcgcagag agcgcatctt
     1801 ttctttagat ggggttcctc attctgatga gcctggaagt aaattaccca gcaaactcca
     1861 gcgtatgtgc tctaagaatg gaaaagaggt cctcaaaact ggaagtcctc agccaacaga
     1921 aaaggccagc agcacaggtg atctgcctct cagggtacag cggcttggtt ccctggtggg
     1981 tcacggagag acagaaaatc acctgcgtct caaacctctt aagatgggtt cagagactca
     2041 cacccagtca gcaggagaac tggagcatcg cagaaagctc tcagcgcctg ctccagtacc
     2101 tggcccaagt ctcaacctat tgcctggttc tagtagccga ccagaggaca atctccctgc
     2161 taaattgcgc agaatgaacg cagaccactg agacccagct ttcttgtaca aagttggcat
     2221 tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa
     2281 aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc
     2341 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     2401 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     2461 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     2521 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     2581 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     2641 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     2701 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     2761 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     2821 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     2881 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     2941 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     3001 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     3061 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     3121 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     3181 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     3241 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     3301 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     3361 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3421 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3481 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3541 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     3601 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3661 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3721 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3781 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3841 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3901 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3961 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4021 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtt
//
