LOCUS       sequence-343336-        3609 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 3609)
  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 3609)
  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-kcnj16
FEATURES             Location/Qualifiers
     source          1..3609
                     /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(1734..1833)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1850..1869)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1851..1869)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1874..1890)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1874..1890)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1992..2011)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2003..2812
                     /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(2070..2089)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2959..3547
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3448..3467
                     /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 agcaggctcc gcggccgccc ccttcaccat gaattcaggt aaggtgcagt actctggagt
      721 gaagagcagt gacaatattc aaagtgtggt cactgacgac agctgttact ccaagaagaa
      781 gcgatatgtg agaaaagatg ggagctgtaa catggtaccc cggcatgttc cagaaaagtg
      841 gcttctgtgg gtcactgaca tcttcactac cttggtggag atccgctgga gagtcatgct
      901 cttgacattt gccctctcct acatcttatc ttggctgttt tttggaattc tattctggat
      961 cattgcatta acacatggcg atatgaaaaa ccccaacaat gagccatgcg tttatgaggt
     1021 tcggagtttt acagctgctt ttcttttctc attagagacc caaaccacta ttggttatgg
     1081 ctttcggggg atgtctgaga actgcatgat cgctatcatc gtagttacca tccaagatgt
     1141 cattagtgtg ttcattgaca cttttgtcat tggcattgcc gttgccaaga tggcgtctgc
     1201 ccgtaaaaga gcccaaacag ttggattcag caagtgtgca gtgatcagta tacatgatgg
     1261 tcacctatgc ctatcatggc gggtcgggga cttccggagg aaccatatgg tagaaggaac
     1321 ttctaatgca cagctaatag agttccaagc aaacaacaac acagggaaag ttgacatcac
     1381 atacaaagac ctcatcattg aagagaggag cgtgatcttg gcaaccccaa ccactataat
     1441 ccataaaata ctccctggaa cgcctctata caagatgagc ttaaaggatc taagaaaaga
     1501 gagcttcgaa ctagtggtct ctttcactta cacagacgat tgcacgggca tccttcacca
     1561 gactcgcaca tcttacactc caagcgagat tctgtggggc cagaactttc aggagatgat
     1621 cagggtcacc cgcaagaact acagagtgga ctacgcccta ttcaaccaca cggttaaagt
     1681 cctggtgcca gaattaagcg cagaggagta tgacctaaag ggtgggcgcg ccgacccagc
     1741 tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca
     1801 ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga
     1861 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     1921 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     1981 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt
     2041 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     2101 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
     2161 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     2221 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     2281 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
     2341 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     2401 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     2461 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     2521 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     2581 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     2641 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     2701 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     2761 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     2821 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc
     2881 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     2941 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3001 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3061 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3121 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3181 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3241 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3301 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3361 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3421 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3481 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3541 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3601 tcacatgtt
//
