LOCUS       sequence-342840-        3716 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 3716)
  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 3716)
  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-kcnj1a.1
FEATURES             Location/Qualifiers
     source          1..3716
                     /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"
     promoter        559..577
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     559..576
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    592..691
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1841..1940)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1957..1976)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1958..1976)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1981..1997)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1981..1997)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2099..2118)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2110..2919
                     /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(2177..2196)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3066..3654
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3555..3574
                     /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 agtcttaaat ttaggtgaca ctatagatta agctcgggcc ccaaataatg
      601 attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgatgagca atgctttttt
      661 ataatgccaa ctttgtacaa aaaagcaggc tccgcggccg cccccttcac catgactcgc
      721 tcattgagag agtggtttaa agagtacatg tcagagcggc gcgggcgaaa aaaccgcctg
      781 gtcaccaaag atggacgttg caacatcgag ttcggcaacg tgatgcacag aaaccgcttc
      841 gcctacgtcc tcgacttctg gaccaccttc gtggagaccc gatggcggtt catcctcttg
      901 tattttgtgg cgtccttcac cctcagctgg ttcatcttcg gactcatctg gtactggttg
      961 gcaagaaaca atggcgacct gttgtggcag aatccaccac caggtcataa accatgcgct
     1021 tttagcgtta atgatatgac ttatgccttc cttttctcac tggagaccca gtccaccatt
     1081 gggtttggca atatatacgt cactgaccaa tgtcctggag gtgtggcttt gattgtaatc
     1141 cagactatca taggactctt catcaacatc ttctggtgtg gtatagtcat gaccaaaata
     1201 tcccttccaa aaaagagagc caaaaccatc accttcagtg aatctgctgt catctgcccc
     1261 aaaaaaggca ctctttgcct tcaaatccgt gtagccaacc tgcgcaaaac cttaatgatc
     1321 ggcagtcaga tttacgggaa gcttctgagg acaacagtca cgggaggtga gaccgtcatc
     1381 ctggaccagg tcagcattga cttcatggtg gacgccggta aagacaatct gtttttcgtg
     1441 tgtcctttaa ccctgtacca catcattgac aaaagcagtc cgttcttcaa catggctgtg
     1501 gacacgatgc atcagcagga ctttgagctt gtggttttcc tcgatggcat ggctgaatcc
     1561 accagctctg cctgccaggt caggacttca tacatccctc aggagatcat gtggggctat
     1621 gactttctcc cgatcatctc ccgcagcaaa gagggaaggt accgagtgga cttttccaac
     1681 tttgctaaag tggtgcctgt gccaaccgct cattgcaccc actgctacca caatgagccc
     1741 ggaaaccatc tccatctcaa gggcattgac aacaagggct ttgaggtgat cgacattgag
     1801 ccgccaaacg gcaccaaaat gtaaaagggt gggcgcgccg acccagcttt cttgtacaaa
     1861 gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt
     1921 caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc gtattacatg
     1981 gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca
     2041 caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca
     2101 aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg
     2161 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     2221 atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     2281 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     2341 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     2401 gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     2461 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     2521 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     2581 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     2641 aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc
     2701 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     2761 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     2821 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     2881 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg
     2941 taacactggc agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc
     3001 cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3061 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3121 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3181 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     3241 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3301 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3361 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3421 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     3481 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3541 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3601 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3661 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgtt
//
