LOCUS       sequence-413265-        3498 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 3498)
  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 3498)
  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-kcnk6
FEATURES             Location/Qualifiers
     source          1..3498
                     /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(1107..1206)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1223..1242)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1224..1242)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1247..1263)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1247..1263)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1365..1384)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1376..2185
                     /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(1443..1462)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2332..2920
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2821..2840
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3074..3091
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3250..3277)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3369..3455)
                     /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 tcaacggcgt tcaggtcctg
      181 ctttatgctc atcggcttca ttttggcgta ctttacatat ctgttattgg gagcgctcgt
      241 tttctcagcc atcgagcggc ccgtcgagga gtccctgaaa gcggacctga gctccctgaa
      301 agccgagttc ctcaacctaa gctgcgttaa cagcacggct ctggaaacct tcttagaaag
      361 agttttaaaa gctaataaat atggcgtgtc ggtgcttgaa aatgcttctc ttcgtacaaa
      421 ctgggatttg gcgtcttctc tgttctttgc caacacgatg gtgacgacag tgggatatgg
      481 ccacacgacg ccactctcag acgccggtaa agccttctcc atcgtgtacg ctctgatcgg
      541 cgtcccgttc accatgctgg tgctgacagc gtgtgtgcag cgcctcatgc acccgctgac
      601 ctacaggccg atctccgcat gccagcggcg ggccggtctg cagcaacggt cagccagcgt
      661 ggtgcacttc atcgtcctgc tgtttctggt ggtgctctgc ttcttcgtgg tgccgtcgct
      721 ggtgttcagc gccattgaag agacctggtc tttcctggat gccttctact tctgcttcat
      781 ctctctctgc actataggac tcggggactt tgtgcctgcg gaaaaaccag gacagagtct
      841 cagagctttg tacaagatat cagtcatggt ctacctgttt gtgggattga tggtgatgtt
      901 cctggtcctc cgcacatttc acaagcttgc tgatgtttac ggctggacgg ctttcttcca
      961 cctgccgagc tgcgaggaag acgaggagga taaagagcca ataatagaag cagggcctga
     1021 agacgattcg ccggaggctg agaaggcctc aattaagcct ctggatccca gctcgcaagt
     1081 ctcctacaac tccataaata gataagaccc agctttcttg tacaaagttg gcattataag
     1141 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     1201 tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc
     1261 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     1321 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     1381 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata
     1441 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta
     1501 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     1561 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     1621 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa
     1681 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     1741 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     1801 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     1861 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     1921 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     1981 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     2041 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     2101 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     2161 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag
     2221 cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta
     2281 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     2341 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     2401 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     2461 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     2521 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     2581 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     2641 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     2701 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     2761 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     2821 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     2881 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     2941 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     3001 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     3061 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     3121 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     3181 gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc gctagccagg aagagtttgt
     3241 agaaacgcaa aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt
     3301 tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct
     3361 cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag
     3421 gcccagtctt ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc
     3481 gttaacgcta gcatggat
//
