LOCUS       sequence-421936-        4060 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 4060)
  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 4060)
  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: pENTR-D-sp6-kcnk4b
FEATURES             Location/Qualifiers
     source          1..4060
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1..18
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    34..133
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1627..1726)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1743..1762)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1744..1762)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1767..1783)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1767..1783)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1885..1904)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1896..2705
                     /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(1963..1982)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2852..3440
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3341..3360
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3594..3611
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3770..3797)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3889..3975)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4024..4046
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4038..4055
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4039..4055
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 atttaggtga cactatagat taagctcggg ccccaaataa tgattttatt ttgactgata
       61 gtgacctgtt cgttgcaaca aattgatgag caatgctttt ttataatgcc aactttgtac
      121 aaaaaagcag gctccgcggc cgcccccttg ccaccatgca cggggcaact cttgtattta
      181 tcctcactgg ggtgctcctg tatttggtta tgggggctct gatgttcaat tatctggagg
      241 ccggtcatga agagaaacac tttgaccttc tgcagaccac caaacaagag ttccttagta
      301 actatacctg tgtggaccta gagaatctaa acaagttcat cgatacacta gcagaggtaa
      361 tgggtgttgg agttgacccc accaaaaatg aaaccaattt cagcaccagc tgggacctgg
      421 ccagcgcttt cttcttctgt ggaaccattg tcaccaccat cggttatgga aacatctctc
      481 caaagacatg gtggggtcag ttcttctgta tctgctatac actggtgggc attcccttgt
      541 ttggatttct gttggctgcc gcaggagacc atctggggac gtcgctgagg aatgccattg
      601 ctaaaattga agcgctttta tggaaattta aggtgaaacc agtcatcgtg cgtgtcatca
      661 cctcagtgct gtccatcctg ctgggctgcg tcctcttcat ctttttgcca attttagtct
      721 tccagaagat tgaaaattgg actctgctgg aatctgccta ctttgtggtc atcaccctca
      781 ctactgtggg ctttggagac tatgttgcag gggatgacgt caagggtgac gatggtcgag
      841 aatattggta caagcctctg gtgtggttct gggttttgtt cggtttggcc tactttgtta
      901 ccattctctc catgattggg aactggatct tggtacttac taaaaaaacc aaagctgaga
      961 tggaggggtt gcgagcccat gcctcggact ggactcagaa catccagaat atgtctgtgg
     1021 atttcaacat tgctggaaaa cttgaagatc ccttcaagcg cagcaagaga aagcgccgtc
     1081 acaggcggca caaacacacc ggcagtcagg gtgaagtcgg aaaagatggt agagaggaga
     1141 atgaatctga atccggctcc tactcatcag atgagtctga aggcagctcg gagtcagagt
     1201 cagaggcaac tcaaactgaa cgggtcccgg agggtggagg aagcgttgct gctaaagaat
     1261 tagagaaaga ggaaaaactt cacgccatcc ccaaagatcc catccccaaa gatcccaagc
     1321 ccaaagatcc catcctcaaa gatcctattc tctctcagcc tttggattac tttggggaaa
     1381 atcttgcata cattgatgaa tcctcagatg cagggagcag gaaggtccac ctagaccccc
     1441 tgctggattt accagactca aacccagaca ggcccaaaag gagacgtcaa cggagacaat
     1501 tgagaagaga tcaaccaaaa accaagtacc cccaaaccag tccaattgaa ccagagaata
     1561 gagcgccaaa tggagacatt caacagaaag atccttctac accaaaagtg aagggtgggc
     1621 gcgccgaccc agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt
     1681 gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata
     1741 tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt
     1801 ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac
     1861 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt
     1921 cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
     1981 ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag
     2041 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
     2101 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
     2161 ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc caggtattag
     2221 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
     2281 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc
     2341 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
     2401 atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg
     2461 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
     2521 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
     2581 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     2641 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
     2701 tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg acttgacggg
     2761 acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt
     2821 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     2881 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     2941 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     3001 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3061 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3121 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3181 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3241 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3301 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3361 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3421 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     3481 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     3541 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     3601 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     3661 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     3721 acgcaattaa tacgcgtacc gctagccagg aagagtttgt agaaacgcaa aaaggccatc
     3781 cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc gtcctgcccg
     3841 ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat ttgtcctact
     3901 caggagagcg ttcaccgaca aacaacagat aaaactaaag gcccagtctt ccgactgagc
     3961 ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta gcatggatgt
     4021 tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa
//
