LOCUS       sequence-413285-        3780 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 3780)
  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 3780)
  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-kcnk13a
FEATURES             Location/Qualifiers
     source          1..3780
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     6..28
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     20..37
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     21..37
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    53..152
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1389..1488)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1505..1524)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1506..1524)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1529..1545)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1529..1545)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1647..1666)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1658..2467
                     /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(1725..1744)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2614..3202
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3103..3122
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3356..3373
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3532..3559)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3651..3737)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gttttcccag tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc cccaaataat
       61 gattttattt tgactgatag tgacctgttc gttgcaacaa attgatgagc aatgcttttt
      121 tataatgcca actttgtaca aaaaagcagg cttcaccatg gcatgtagag gctcgtgctg
      181 ttgtggctgt ggtccagtga atgaagataa tgcccggttc atcatgttgc tactcctcat
      241 catcctctac cttctgtgcg gagccgcggt gttttcagcc ctggagcatc cgaaggagaa
      301 actggcgaag gaaaggtggg ctcagaggtt tgagctgttc agccaaaagt acaacctcaa
      361 caaaagtgac ctgaagaact tcttgagaca ttatgaagag gcgaacatgg cagggatacg
      421 ggtggacact ctcagaccgc gttgggattt taccggagcc ttttattttg tcgggactgt
      481 cgtttctacc atcgggtttg gaatgaccac ccccgccacg gtcggtggca aaatcttcct
      541 gatcttttat ggcctcatcg gctgcgcagc caccatttta tttttcaatc tcttcctgga
      601 gagaataata acagtgttgg cgttcgtgct gaagtcgtgc catgaaatcc aacgtcgacg
      661 caagggagtt ttacctcacg acagcagaca ggactcaagg ctaaaccatg aagacagtct
      721 tgctggatgg aagccctcgg tttactgtgt gatgctgatc ttatgcgtgg cggcaataat
      781 catttcttgc tgtgcttcag tcatgtactc ttcaatcgaa ggatggcgat acttggattc
      841 gctttacttc tgctttgtgg cgttcagcac cattggtttt ggggatatgg tcagcagcca
      901 acgagccatt tacgagaacc aaaccgtcta ccgaatttgc aacttcctgt ttattttaat
      961 gggcgtttgc tgcatttact ccctattcaa cgtcatctcc atcgtgatca aacaggtgct
     1021 caactggctc ctgaaaaagc tgaagatacc ttgtcgtcac tgtcacagga ggtctcttcg
     1081 gccccgcagg aacgcagtca tgccgagcca tctccgaaca cgcgtccgca acatatccat
     1141 agacacggac gcagtgaacg acagtgagac ggatggacgc agactgtctg gagagatgat
     1201 ctccatgaga gacttcctgg cggctaataa ggtcaacttg gcgatcatgc agaaacagct
     1261 gtctgagact gccaatggtc atccccggcc gggcggctca acaaacagac acaatggatt
     1321 ctctggtgga gtcggagctt tgggtataat gaataacagg ctagcagaaa ccagcgtgga
     1381 cagataagac ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     1441 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     1501 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     1561 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     1621 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     1681 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     1741 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     1801 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     1861 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     1921 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     1981 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     2041 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     2101 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     2161 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     2221 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     2281 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     2341 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     2401 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     2461 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     2521 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     2581 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2641 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2701 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2761 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2821 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2881 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2941 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3001 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3061 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3121 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3181 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3241 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     3301 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     3361 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     3421 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     3481 caacgcaatt aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca
     3541 tccgtcagga tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc
     3601 cgccaccctc cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta
     3661 ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga
     3721 gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat
//
