LOCUS       sequence-208832-        4450 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 4450)
  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 4450)
  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: Pmo_piwi-1
FEATURES             Location/Qualifiers
     source          1..4450
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     105..121
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     105..121
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        126..144
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        202..220
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2132..2148)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2132..2148)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(2206..2224)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2229..2276
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2283..2300)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2283..2299)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2292..2314)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      2363..2449
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2541..2568
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2727..2744)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2898..3486)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2978..2997)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3633..4442)
                     /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     4356..4375
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     join(4434..4450,1..3)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 ttgtattact gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt
       61 gtgcaatgta acatcagaga ttttgagaca cgggccagag ctgccaggaa acagctatga
      121 ccatgtaata cgactcacta taggggatcc tcaaaaaacc cctcaagacc cgtttagagg
      181 ccccaagggg ttatgtggaa gtaatacgac tcactatagg gcgaattggg taccgggccc
      241 ggccaattct acccgccggg attaggatta ttaccatctt taaaaaactt gacacttatt
      301 ctctttttaa caattatata aattatttgg ggcaaatttt cattttcgta aattgtttcc
      361 ataattttta aaacagcatc agtttcgtac tttttagtga aatacaattg agaatctcca
      421 acaccgtctc gataaataat aaacctttgt ggcaaagtgt taaatttatc tttaaatgta
      481 tttatggctg cgatgaaatt ttgactcaag ttatcatgat attcattctt gccttttggt
      541 gaatttacaa aactcaaata ttgagaaaat ttcgaactga tactgaatac agatgcttga
      601 attgatcgct tgccagtctt gctatgaaaa gtatccaaac caacaaccat cgtcggtacc
      661 atttttaact caattcccca tggatcgtaa cccaatttag caaaaatttg cataacactt
      721 ttatcagcca caacttttct acgacgatca tctctatttg tcccatttct tacagtcacg
      781 cactgggtta ataatccagt gcttatcgta aagttcttta cacgagcata tgccttatcg
      841 tcaccaataa aaaccaaagc caaatgaacc ttttgaactc taacaaaatc tgttaagcaa
      901 tcttcaattt catttgctct gcaagttgca acagtctcta ttgaataatt aactcttagc
      961 ctacttattt cacgcttaac atcatctata aattgattga attgactagg acttctttcg
     1021 acggctaaaa tagcaaaacg atgctgagca ccctttggaa tatcaaattt aacttcaccg
     1081 aatttccaat catctggaga tttggcatta atacgttgtt gcccagaaaa tacatcaacc
     1141 ccatttagtt ctcgccccat aattgttatt ggttgattat ctaattgaac tccccaattt
     1201 ttcataaatg gagccgcctt gccttcatca gtccaactac aaaaatcact aatatctttt
     1261 aatcgttctc gtggttctct tttaagaact gatcctagac tcttttgcaa attaatgttc
     1321 gatctctcac tatccgaaaa gccacacaaa aagcacaatt cgccaggaat agataaactt
     1381 tgatcagctt cctttggtgc ctcgggttca ttttcttgat tttctttttg ttgaatagcc
     1441 actcttctca atcttgatat gacaaatggt tgctttccat tattaagcgt tatgttgtat
     1501 ctttgcttaa aataattggc atatgaaatc tttgtaccat ttaattctat ttcgtcatta
     1561 acatccatct ccttaatatc ggaaattctg tatgttttgt tattatattt ggttataaca
     1621 tctcttccaa tcaacagatc caattgattt ttattataaa tattataaac tgaattttca
     1681 ttcaatactc gattaatgcg ctctaaataa agtaaagttg aagcacttcc atcggaatta
     1741 gttcctcttg acaatgttgt agcaaaaccg cccaatattt taaaattatc tcttttgaaa
     1801 aaagatggcc cacatttgga tgtagattca ttactattcg aacccaagaa aaaagctcta
     1861 ccaattcgtt cttgtcccat gaacgcttga atgttattga agattatgtt aaccatttga
     1921 taatattctt cggtattttc cacaaatgag gctaaaaaac gaattgatac ttcttcatct
     1981 tttatcgaaa ttgtttctat atcttcttgg tgccactgtg aagatgtaaa aagacggcga
     2041 ccatcgaaac aaaatggccg attttctttc gttgaagcga cgctgtttaa taagtcccgg
     2101 gatggtaatg ccgctagcca tggaaccggt ggatccacta gttctagagc ggccgccacc
     2161 gcggtggagc tcgaattcat cgatgatatc agatctgccg gtctccctat agtgagtcgt
     2221 attacttcct agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctta
     2281 agactggccg tcgttttaca acgtcgtgac tgggaaaaca tccatgctag cgttaacgcg
     2341 agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcggaag actgggcctt
     2401 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc
     2461 ggatttgaac gttgtgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac
     2521 tgccaggcat caaactaagc agaaggccat cctgacggat ggcctttttg cgtttctaca
     2581 aactcttcct ggctagcggt acgcgtatta attgcgttgc gctcactgcc cgctttccag
     2641 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     2701 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     2761 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     2821 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2881 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2941 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3001 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3061 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3121 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3181 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3241 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3301 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     3361 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3421 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3481 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgacgcgtaa
     3541 ctcacgttaa gggattttgg tcatgagctt gcgccgtccc gtcaagtcag cgtaatgctc
     3601 tgccagtgtt acaaccaatt aaccaattct gattagaaaa actcatcgag catcaaatga
     3661 aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt
     3721 aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct
     3781 gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg
     3841 ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta
     3901 tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc
     3961 gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg
     4021 ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc
     4081 gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttt
     4141 ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg
     4201 gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca
     4261 ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac
     4321 aagcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat
     4381 aaatcagcat ccatgttgga atttaatcgc ggcctcgacg tttcccgttg aatatggctc
     4441 ataacacccc
//
