LOCUS       sequence-413243-        3567 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Riboprobe generation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3567)
  AUTHORS   Park SJ, Silic MR, Staab PL, Chen J, Zackschewski EL, Zhang G
  TITLE     Evolution of two-pore domain potassium channels and their gene 
            expression in zebrafish embryos.
  JOURNAL   Dev Dyn. 2024 Jan 25. doi: 10.1002/dvdy.690.
  PUBMED    38270285
REFERENCE   2  (bases 1 to 3567)
  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. 
            2024 Jan 25. doi: 10.1002/dvdy.690."
COMMENT     Sequence Label: pDONR221-kcnk1a
FEATURES             Location/Qualifiers
     source          1..3567
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(936..1035)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1052..1071)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1053..1071)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1076..1092)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1076..1092)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1194..1213)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1205..2014
                     /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(1272..1291)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2161..2749
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2650..2669
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2903..2920
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3079..3106)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3198..3284)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3333..3355
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3347..3364
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3348..3364
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3380..3479
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ggcgtatgtt ttatatctta tcttcggggc tttagtgttt tccgcggtgg agctgccgta
       61 tgaagagcag cttcgacagg agcttcggac agtgaaacag caatttctgg aggacaatga
      121 atgtctgtct aatgacagac tggaagagtt tctgatcaaa gcgttagaag cgagtaatta
      181 cggtgtgtcg gttcttaata atgcctccag taactggaac tgggatttca cttcagcgtt
      241 gttcttcgcc agcactgtgc tttccacgac aggatatggg cacactgtcc ctctatcaga
      301 tggaggtaag gccttctgca tcatttactc agtggtggga ataccgttca ccctgctgtt
      361 cctgacggcg gtggtccaga ggatcatgga gttcagcact cgcaggccca tagagttcct
      421 gcaccggcga tggggcacct ccaaacctct gctggctgcc atgcacgcca cactactggc
      481 cattataact gtctcctgct tcttcctcat acctgccatc atcttctctg tgctggagga
      541 ggagtggaac ttcctggagt ccttctattt ctgctttatc tctcttagta ccattggctt
      601 aggggactat gtgccaggag agggttacca tcagaggttc agagaacttt ataaactggg
      661 cattactttc tacctgatat tgggcttgat cgctatgttg gtggttctgg aaaccttctg
      721 tgagcttcag cagctgaaga agcttaggaa aatgttctac ctgaggaaac aaaaaacgga
      781 ggaccagcta aacatcgtag atcacgatca cctctctttt gcctctgtgt ccgaccaggc
      841 agcctcattc agagaggata agactgaact gtttcctgat gatattgtgt ccggctcacc
      901 tactgctacc tctaatggat cactggagcg atgagaccca gctttcttgt acaaagttgg
      961 cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
     1021 taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat
     1081 agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     1141 taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg
     1201 tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc
     1261 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta
     1321 tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt
     1381 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct
     1441 tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat
     1501 ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt
     1561 tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt
     1621 taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt
     1681 tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga
     1741 aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact
     1801 tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg
     1861 aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc
     1921 ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt
     1981 gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca
     2041 ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa
     2101 cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2161 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2221 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2281 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     2341 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2401 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2461 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2521 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     2581 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2641 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2701 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2761 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     2821 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     2881 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     2941 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     3001 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga
     3061 agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc
     3121 ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc
     3181 aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata
     3241 aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc
     3301 tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc
     3361 cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga cctgttcgtt
     3421 gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa aagcaggctt
     3481 caccatgctt cagtgtctgt cgggaagttc gtgtatgcgt gtcattgacg gttacaagtc
     3541 tacgtggtat ttcttagttc tggtgct
//
