LOCUS       sequence-413286-        3718 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 3718)
  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 3718)
  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-kcnk18
FEATURES             Location/Qualifiers
     source          1..3718
                     /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(1327..1426)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1443..1462)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1444..1462)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1467..1483)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1467..1483)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1585..1604)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1596..2405
                     /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(1663..1682)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2552..3140
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3041..3060
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3294..3311
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3470..3497)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3589..3675)
                     /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 tccgtgtcac aaaagagaca
      181 gtttacgggg agatgctcga cactcttctg gcgtttgttt cctcatgttt tcctaatttt
      241 atcactcgtt ttatacgcgg ttttaggagc gcttgtcttc agagcgattg agtacacgaa
      301 tcctcgaaat gagtcggagg agatcctgtc cattgtgcaa aaagtgatgg agatcgttca
      361 gaatcacaca gacgcctcag agcagaagca cctaatcaat aaagcaaaat acattttgga
      421 tgattactgt tatgagaagg aacgtgatca cggatggacc ttctttgctt cacttttctt
      481 ctgctgtaca gtgttcacta ctgtaggtta tggacgaatt tatccgttga caagtaaagg
      541 gaaagtggca tgtgtcctgt atgcgatggt gggcatccct ctgatgctgc tggtcatctc
      601 ggatgtgggc gatctcttag ctgttctcct ctccaaagca tatactcgcc ttaacctgtt
      661 cttcagaaga tggataggac atcagtcatg gagacttcag agccacgaaa agacatcagc
      721 tctcccacaa gttcaagctg atactgacgg cacctataag ttcaatcagg atgtcgtggt
      781 gctcgagaca acaaacaatc aacaggtgat acaaactagg tcttccatca ggcgcgggtc
      841 ctttcagctc cgtaacaaca aagagatctt tgaccgcatt attgtaaggg agagcttcag
      901 gatcaaagga actctttcaa aatcctgttc atgcccagaa ctggaccgtg tgccaacccc
      961 aaaagatgag ttattcaacg acatcggaca ggagatggag cagttggacg tccctttgct
     1021 ggtcatcttg ctgatggtgt ttgcctacat ggtgatctgc agtcagattc tgaaatgctg
     1081 ggagaagcag atggaccatt ctgatgcctt ttatttcacc ttcatcaccc taaccaccat
     1141 tggctttggc gacattgtgc ctgaacaccc aaaattcttc atggttactt ttttgttcat
     1201 catcacaggc atggccatta tgagtatggc ctttaaactt ggccaatcgc aaattgtttg
     1261 cttttacagg cggtgtataa agagtctaag catgggcaag gtaagaattc acaatgactt
     1321 aaacagaccc agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt
     1381 gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata
     1441 tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt
     1501 ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac
     1561 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt
     1621 cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
     1681 ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag
     1741 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
     1801 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
     1861 ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc caggtattag
     1921 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
     1981 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc
     2041 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
     2101 atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg
     2161 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
     2221 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
     2281 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     2341 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
     2401 tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg acttgacggg
     2461 acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt
     2521 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     2581 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     2641 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     2701 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     2761 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     2821 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     2881 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     2941 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3001 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3061 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3121 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     3181 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     3241 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     3301 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     3361 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     3421 acgcaattaa tacgcgtacc gctagccagg aagagtttgt agaaacgcaa aaaggccatc
     3481 cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc gtcctgcccg
     3541 ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat ttgtcctact
     3601 caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt ccgactgagc
     3661 ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta gcatggat
//
