LOCUS       sequence-343132-        3759 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 3759)
  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 3759)
  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-kcnj6
FEATURES             Location/Qualifiers
     source          1..3759
                     /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(1884..1983)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2000..2019)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2001..2019)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2024..2040)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2024..2040)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2142..2161)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2153..2962
                     /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(2220..2239)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3109..3697
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3598..3617
                     /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 accatggagc aggatgtcga gagcaccgtc acggttcgac aacccaagct
      721 gcccaagcag gcgcgggacg acatccccaa accgctgcag gagcgggagc gcgccaagca
      781 gaagacccag cgctacgtcc gcaaagacgg caagtgcaac gtgcaccacg gaaacgtgcg
      841 ggagacctac cgctacctga cggacatctt caccacgctg gtggacctca agtggaggtt
      901 caacctgttc atcttcgtgc tggtctacac cgtcacctgg ctgttcttcg gcttcatgtg
      961 gtggctgatc gcctacattc gaggagatct ggagcacatc ggagacaacc agtggactcc
     1021 ctgcgtcaac aacctgaacg gcttcgtgtc cgcattcctg ttctcgatag agactgagac
     1081 caccatcggc tacggccacc gcgtcatcac cgacaagtgt ccggagggca tcgtgctgct
     1141 gctggtgcag tcggtgctgg gctccatcgt caacgccttc atggtgggct gcatgttcgt
     1201 caagatctcg cagcccaaga agcgcgccga gacgctggtc ttctccacca acgccgtcat
     1261 ctccatgcgg gacggccggc tctgcctgat gttccgcgtg ggagacctca gaaactcgca
     1321 catcgtggag gcgtccatta gagccaagct gatcaagtcc aaacagacca aggagggcga
     1381 gttcatcccg ctcaatcaga cggacataaa cgtgggctac gacaccggag acgaccggct
     1441 cttcctggtg tctccgctca tcatctgcca cgagatcaac cagcacagcc ccttctggga
     1501 gatctccaga gagcacatgg ccaaggagga gctggagatc gtggtgatcc tggagggcat
     1561 ggtggaggcc acaggcatga cgtgtcaggc gcgcagctca tacgtggcca gcgagatcaa
     1621 gtggggttac cgcttcacgc cagtgctgac gctggaggac ggcttctacg aggtggacta
     1681 caacagcttc cacgagatct acgagaccaa cacgcccagc tgcagcgcgc gagagctggc
     1741 cgagatgacg gcgcgcgccc gcctgccgct cacctggtcg gtggccagca aactgagcca
     1801 gctggagccg caggagccgc cggaggagct gacggagagg aacggagacg tcgccaacat
     1861 ggagagcgag tccaaagtgt aggacccagc tttcttgtac aaagttggca ttataagaaa
     1921 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     1981 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     2041 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     2101 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     2161 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     2221 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     2281 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     2341 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     2401 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     2461 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     2521 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     2581 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     2641 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     2701 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     2761 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     2821 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     2881 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     2941 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     3001 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     3061 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3121 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3181 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3241 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3301 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3361 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3421 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3481 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3541 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3601 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     3661 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3721 acggttcctg gccttttgct ggccttttgc tcacatgtt
//
