LOCUS       sequence-413326-        3789 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 3789)
  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 3789)
  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-kcnk13b
FEATURES             Location/Qualifiers
     source          1..3789
                     /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(1398..1497)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1514..1533)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1515..1533)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1538..1554)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1538..1554)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1656..1675)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1667..2476
                     /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(1734..1753)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2623..3211
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3112..3131
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3365..3382
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3541..3568)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3660..3746)
                     /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 gcttgccggt ctggatgctg
      181 ctgcaacagc atcggcagct tcaatgaaga caatgcgagg tttctgatgc tggctttgct
      241 gatcatcatc tacttgctgt gcggggcagc ggtgttttcg gcgctcgagc agccgaaaga
      301 gaagctggcc aaggagagat gggctcagag gatcgagcag ttcacgaaca agtacaacct
      361 gagtcaagac gacctgaaga acttcttgag gaactatgag gaggccaatg tggcgggaat
      421 ccgcgtggac gcgacccgcg cgcgctggga ttttgcagga gctttttatt tcgtcggaac
      481 tgtggtttca actatcggtt ttgggatgac gacacccgtc actattgctg ggaagatatt
      541 cttgatcttt tatggactaa tcggttgtgc cgccaccatt ttgttcttta acctcttcct
      601 ggaacgtgtc atcaccgtga ttgcatttgt tttgaaattc tgccatgagc gcagagaaag
      661 tcgaaaagca ggcccgactc aaaactgccg tcgtccttcc accgacaaca gagacaggcg
      721 cacagacagc ttggccggct ggaaaccttc agtgtactgt gtgatgctca tcttgggagt
      781 tgctgccatc ttggtctcct gctgtgcatc tgctatgtac tcggcagccg agggttggga
      841 ctacctggat gccctctatt tctgttttgt agctttcagc actattggtt ttggggatat
      901 ggtgagcaac cagcgtgaga tatatgaggc tcaggtggcg tatagggtgg gcaacttcct
      961 tttcatccta acaggggtgt gctgcatcta ctcgctgttc aacgttattt ccattgtaat
     1021 aaaacaagtt ctcaattggc ttcttcggag attagagact ccctgccatt gtccaggccg
     1081 tgggaatcgc catccaagga ggaatgctgt cgtccctggc cacatgagat cccggaggga
     1141 aaactctatc gagactgatg cagtcaatga cagtgaggca gatggacggc ggatgtctgg
     1201 agagatgatc tccatgaagg acttcctggc tgccaataaa gttaatctgg ccatcatgca
     1261 gaagcagctg tcagagatgg ccaatggaca ccctcgccag tcagggagca gctcacggca
     1321 caatgaattc tccggaggag tcggagcact gggaatcatg aacaaccgac tggctgagac
     1381 cagtgtagac agataagacc cagctttctt gtacaaagtt ggcattataa gaaagcattg
     1441 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     1501 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     1561 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     1621 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     1681 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     1741 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     1801 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     1861 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     1921 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     1981 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     2041 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     2101 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     2161 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     2221 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     2281 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     2341 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     2401 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     2461 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     2521 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     2581 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2641 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     2701 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2761 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2821 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2881 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     2941 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3001 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3061 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3121 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3181 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3241 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     3301 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     3361 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     3421 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     3481 cagtgagcgc aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca
     3541 aaaaggccat ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg
     3601 cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga
     3661 tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
     3721 tccgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct
     3781 agcatggat
//
