LOCUS       sequence-342883-        4170 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 4170)
  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 4170)
  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-kcnj3b
FEATURES             Location/Qualifiers
     source          1..4170
                     /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(2295..2394)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2411..2430)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2412..2430)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2435..2451)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2435..2451)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2553..2572)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2564..3373
                     /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(2631..2650)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3520..4108
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4009..4028
                     /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 cgttgaggaa aaaatttggg gatgattatc aggtggtgac
      721 cacctcgtcc agcggcggcg ggttcaacca agcggccccc gagaagaaaa ggaaacgtca
      781 gcgctttgtg gacaagaacg gccgctgtaa cgtccagcac gggaacctgg gaggagagac
      841 gagccggtac ctgtccgacc tgttcaccac gctcgtggac ctcaaatggc gctggaacct
      901 tttcatcttc atcctcacat acacggtggc gtggctcttc atggcttcta tgtggtgggt
      961 cattgctttt atacgcggag acatgaacca ggcccacgac gacaagtaca cgccgtgcgt
     1021 ggccaatgtg tacaactttc cctccgcttt tctgttcttc atcgagaccg aggccactat
     1081 cggatacggt taccgctaca ttacagacaa gtgtccggaa ggcatcattc tcttcctctt
     1141 ccagtccatc ctaggctcta tagtggacgc ttttctaatc ggctgcatgt tcatcaagat
     1201 gtcccagccg aagaaacgcg cggagacgct catgttcagc gagcacgcgg cgatttccat
     1261 gcgtgacggc aaactgaccc tgatgttccg cgtggggaac ctgcgcaaca gccatatggt
     1321 ctccgcgcaa atacgctgca aactgctcaa atcccgtcag actcccgagg gcgagttttt
     1381 gcctctggat caacttgagc tggatgttgg gttcagtacc ggagctgacc agcttttcct
     1441 agtgtctcct ctgacaatct gccatgtgat cgactccaag agcccattct acgacctctc
     1501 gctcaggtcc atgcaaacag agcaatttga gattgtggtt atcttagaag ggatcgtgga
     1561 gaccaccggt atgacatgcc aggctcgtac ttcctacaca gaggacgagg ttctgtgggg
     1621 tcaccgattc tatcctgtca tttctctgga ggagggattc ttcaaggttg actattcgca
     1681 gtttcatggc acttttgagg ttcccacacc accgcacagt gttaaagaac aggaggagag
     1741 tctgctgctg tcgtcccccc tgacagcccc ctctctgtgc cacagcgcag gtggggttac
     1801 agagggcagt ggcacactgg aaggtctgga gcttcaaagc gatgcagaaa ctggctccat
     1861 caatattgca accccaatgc caaccaacaa actgcctgcc aaacttcaga agctcacagg
     1921 acgggaggtc cccattcggg atggaatgcc ccgtaagctt ctgcgcatga gctcggaaat
     1981 cacctacaac ctgggcgacc tgcccgtcaa actccagcga atcagctcag ctccgggcgt
     2041 ggcagaagag aggcttcagc caatcagttc catgctgagc cccacagagg acagactggc
     2101 tcccaagctg ggcttaatag gtggagctga aactatcagc caatcagtga cagatctgcc
     2161 gccaaaactg cagaggttgg ctggcggggt cggggtggga ggagtcggaa tcggaggacg
     2221 aatggatggt catctgcctc ccaaactaag aaaaatgaac tctgagaggt tcacaggctt
     2281 gagtttaaaa tgagacccag ctttcttgta caaagttggc attataagaa agcattgctt
     2341 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     2401 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     2461 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
     2521 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     2581 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     2641 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
     2701 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     2761 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     2821 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     2881 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     2941 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     3001 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     3061 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     3121 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     3181 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     3241 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     3301 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     3361 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     3421 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     3481 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3541 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3601 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3661 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3721 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3781 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3841 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3901 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3961 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4021 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4081 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4141 ggccttttgc tggccttttg ctcacatgtt
//
