LOCUS       sequence-413242-        3480 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 3480)
  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 3480)
  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-Topo-kcnk17
FEATURES             Location/Qualifiers
     source          1..3480
                     /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             739..756
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     protein_bind    complement(1089..1188)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1205..1224)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1206..1224)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1229..1245)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1229..1245)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1347..1366)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1358..2167
                     /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(1425..1444)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2314..2902
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2803..2822
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3056..3073
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3232..3259)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3351..3437)
                     /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 ctccgcggcc gcccccttca ccatgctctc
      181 catgttttca ggaaggttat gtcagtttct cagtgtaacc cggttacctt ctatcctttt
      241 tcttggagtt ttttatgtcg cttatgttct tgtcggaggg ctggtattct ggaagctaga
      301 aggatggtat gtgttgcaac agattgctct tttgaaagag aaaagactgg aactgcttga
      361 gaagtacccc tgtctgggtc agaatggcct cagtgaacta gcacagatga tcaaatctgc
      421 ttccctgaga ggcttgagtc cagacagcaa cgacactgca gatggcctct ggaaatttac
      481 cagttcatct gtgtttgcgg cgacagtggt cacaactatt ggctatggga atatagttcc
      541 gctgacaaca gctggtcaaa tattctgtgt gatgtttgct cttttcggca ttcctctgaa
      601 tgttgtagtc ttgaacagag ttgggaaata catgctggca attgaaagac atttttgcaa
      661 ttttctggag aaaaaaattg atagagggaa atgtgtacgg atatcaattc atagtatatc
      721 ctttgtgagt tcagcgttct tgtacctcgt ggttcctatg ctgctcttta aagagtatga
      781 gggatggagt tatgctgaag ccatttatta ctgcttcata acactcagca ctattggatt
      841 tggagattac gttgcagatc acaacccagc aataaattac cctgagtggt acagctgttt
      901 gatggctgtc tggatattct ttggcctggc ttggcttgcg cttctagtca atcacactat
      961 tgatttactg gaaagtctga acgcttacct gagaggaagg cagaatggac aatcacaaat
     1021 gaatgagact gaagaaaagc cagaaaaagg actgaaagag gaagaaacat gaaagggtgg
     1081 gcgcgccgac ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     1141 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     1201 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     1261 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     1321 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     1381 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     1441 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     1501 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     1561 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     1621 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     1681 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     1741 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     1801 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     1861 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     1921 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     1981 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     2041 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     2101 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     2161 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     2221 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     2281 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2341 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2401 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2461 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2521 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2581 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2641 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2701 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2761 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     2821 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     2881 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     2941 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     3001 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     3061 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     3121 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     3181 caacgcaatt aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca
     3241 tccgtcagga tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc
     3301 cgccaccctc cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta
     3361 ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga
     3421 gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat
//
