LOCUS       sequence-208222-        4526 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In situ hybridization and RNAi studies in planarians.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4526)
  AUTHORS   Accorsi A, Williams MM, Ross EJ, Robb SMC, Elliott SA, Tu KC, 
            Sanchez Alvarado A
  TITLE     Hands-On Classroom Activities for Exploring Regeneration and Stem 
            Cell Biology with Planarians
  JOURNAL   The American Biology Teacher 79(3):208-223. Mar 2017.
REFERENCE   2  (bases 1 to 4526)
  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: "The 
            American Biology Teacher 79(3):208-223. Mar 2017."
COMMENT     Sequence Label: Ddo_piwi-1
FEATURES             Location/Qualifiers
     source          1..4526
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(24..71)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        76..94
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     152..168
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     152..168
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(2156..2174)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2232..2250)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2255..2271)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2255..2271)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2373..2392)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2384..3193
                     /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(2451..2470)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3340..3928
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3829..3848
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4082..4099
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4258..4285)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4377..4463)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4512..4526,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4526,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagcaaaaaa cccctcaaga cccgtttaga ggccccaagg
       61 ggttatgcta ggaagtaata cgactcacta tagggagacc ggcagatctg atatcatcga
      121 tgaattcgag ctccaccgcg gtggcggccg ctctagaact agtggatcca ccggttccat
      181 ggctagcggc attaccatcc cggcgaggag tactaccaaa tggtgaatat cgtattcaat
      241 aacgtacagt gtttgatggg acaagagaaa atcggtcgcc agtattttat ttcatcgaat
      301 atcgattctg tgtcgttaaa agagggtccg agtttcttta gaatggaaaa tttcaagatt
      361 ttaggtggat tttctacgac aatcacccga ggaaccaatt cggatgggac tgcgtcgacg
      421 ctgttgtatt tggagcggat caatcgtgtt ttgaatgaga gtagtgtgtg tagcgtgtat
      481 cgacgggatc aaatcgacgc gctgatcggg cgagatatca tcacgaaata taacaataaa
      541 acgtatagga tttccgatat caaagaaatg gcaattgacg atgaaatcga gttgggcggg
      601 aagcggattt cctatcgcga gtacttcaaa cagcggtata atattgcttt gaagactgac
      661 aagcaaccat ttgtgatttc ccggatgaaa cgatcgatcg cgccgaattt gaggaacaag
      721 aataaagcgg aaggcgcggc ggaagaggga gacgttccac cgagagagcc cgaccagagt
      781 ctatcgattc ccggagaatt gtgttatttg tgcgggttca gcgacagcga gcgcgctaat
      841 atcaatctcc agaagagcat gggaggcgtt ttgaagagag aaccgcgcga gagactcaac
      901 gacatccggg atttctgtaa gtggacgagc aatggcgcac agactcgcga attcatgacg
      961 agttggggaa tgaacattga cgagcagccg atcacgatcc gcggtcgaga aatgccgccg
     1021 gtcgatatct tctcgatgga ccagaagatc aacaccaaga ttcctgacga ctggaagttt
     1081 ggagaagtga gattcgacat ccctaaaggc acggcgcatc gtttcggcgt gctcgtggtc
     1141 gaccgaaacc ctgcgcattt taacgcgttc attgaggacg tgaagcgcga gatcggccgc
     1201 ctccggatca actacacgat cgaggccgta tcgtcatgtc gtggcaacga ggtcgaggac
     1261 gctctggccg agttcatcaa gatcaacaag gttcacatgg ctctcgtgtt catccccgac
     1321 gacaaagtct acgccaaggt caagaacttc accatgtcaa ctggcctgct gacccaatgc
     1381 gtcacccagc gcaacggcag caaccgcgac gaccgccgca gaaaggtggt ggccgacaag
     1441 tccgtcatgc agatgttctc taagctcggc tacgatccat ggggcatcaa cctcaagatg
     1501 gcgcccacca tgatcgtggg actcgacacg ttccactcga aaacgggcaa gcggtccgtg
     1561 caggcgtccg tgttctcgat tagcgccaag ttctcccagt acatcagctt cgtcaacagc
     1621 tcgaagggca agaacgagtt ccacgagaac ctggccaaga acttcctgac ggcaatcaac
     1681 acgttccaga gcaagttcaa cacgatgcct cagcgcttca tcatttaccg cgacggcgtg
     1741 ggcgactcgc agctcgcctt caccaagaag tacgagacgg aggccgtcat gaagatgctc
     1801 gagacgatct acgagaacca gccgctgccg cagatcctct acgtgatcgt caagaagcgc
     1861 atcagcgtca agttcttcaa ggacggcggc aaccccaatc cgggcacgat cgtcgacgag
     1921 aaggtcgtca agccgaactt ctacgaattc tatctcgtgt cccagaaaac taccaaggga
     1981 acggcgtcac cgaccaacta caacgtgctc atggacacca agatgacgaa caagaagacg
     2041 aacgaggtgg cggcgatggc gcccagcgtg ctgcagcaaa tcacgtactc gctcacgcac
     2101 ctctacttca actggatggg cgggtagaat tggccgggcc cggtacccaa ttcgccctat
     2161 agtgagtcgt attacttcca cataacccct tggggcctct aaacgggtct tgaggggttt
     2221 tttgaggatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     2281 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
     2341 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     2401 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     2461 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
     2521 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     2581 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     2641 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     2701 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     2761 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     2821 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     2881 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     2941 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     3001 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     3061 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     3121 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     3181 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     3241 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     3301 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3361 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3421 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3481 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3541 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3601 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3661 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3721 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3781 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3841 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3901 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3961 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     4021 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     4081 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     4141 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     4201 tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa
     4261 aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt
     4321 cctgcccgcc accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt
     4381 gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc
     4441 gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc
     4501 atggatgttt tcccagtcac gacgtt
//
