LOCUS       sequence-413261-        3579 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 3579)
  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 3579)
  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-kcnk7
FEATURES             Location/Qualifiers
     source          1..3579
                     /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"
     CDS             881..892
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     protein_bind    complement(1188..1287)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1304..1323)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1305..1323)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1328..1344)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1328..1344)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1446..1465)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1457..2266
                     /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(1524..1543)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2413..3001
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2902..2921
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3155..3172
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3331..3358)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3450..3536)
                     /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 gtcgcgtacg gcttgtttat
      181 cgtcatgggc gctgttgttt tgatggtact tgagcaacct gaggagaatt tgctggttca
      241 agaggtccgt gaactgaaag ctcgttttct ggcagataac ccgtgtgttg aggagcgaag
      301 tttggatggt ttgttaatgg acgtgctgtc tgctagtaaa cgcggtgtgg ctgcactcca
      361 agcagacagt gacgagtgta atttcgactt tacgtcgtcg ctgtttttcg tcatcacctt
      421 cctcacgacc acaggttatg gcaccaccgt gcccctttca gacgagggcc gtgtgttctg
      481 tgtggtttat tgtctcgtgg gaatccctct caccatgctg ctgctgtcct gcctcactca
      541 tgcccttctg ccccgagtaa ctcacacccc tatccagaac ttgcagctgt tctggggtct
      601 gtcccgtagc cacgctgcgc tcctgcactg cagcatcctc ggcttttgta cagccgcact
      661 cttcttcctg ctgcccgctg ctgctctctg cctgctggag gacgactgga catacctgga
      721 gtcgctgtat ttttgtttta tctcactgag cacaactggt cttggtgact acctgcctgg
      781 caaaatacaa aaccaagcag tgcggcaggg gctggagttc gtcacctcct gttatctgtt
      841 gctcggactg atcgttttgc tcgtggttct ggaaagcttc tgggagctgc agcagtttca
      901 ggcagtcctg cgtttctttt ctggcccgcg gctcagtgag ctaaatggac tgagtttaga
      961 tgagctcgtg ctcaccggag acatgaccgg tccagaggag gaacctcaat acacccttcc
     1021 aatttccacc atctcccctg ctttctcaga ctcacctgcc actccaacaa tagagctgct
     1081 gccagtcttt ggcctgccat ccataccagc ctccaaagat tatgaccact cttcactacg
     1141 gccaaacccc acctcagatc tgtgtcattc cacatcctct aactaagacc cagctttctt
     1201 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     1261 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     1321 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     1381 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     1441 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
     1501 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     1561 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     1621 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     1681 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     1741 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     1801 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     1861 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     1921 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     1981 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     2041 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     2101 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     2161 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     2221 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     2281 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc
     2341 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     2401 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2461 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2521 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     2581 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2641 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     2701 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     2761 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     2821 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     2881 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     2941 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3001 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     3061 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     3121 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     3181 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     3241 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac
     3301 cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc
     3361 ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc
     3421 ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac
     3481 aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt tatttgatgc
     3541 ctggcagttc cctactctcg cgttaacgct agcatggat
//
