LOCUS       sequence-208117-        4811 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 4811)
  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 4811)
  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: Gsp_piwi-1
FEATURES             Location/Qualifiers
     source          1..4811
                     /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(2441..2459)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2517..2535)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2540..2556)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2540..2556)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2658..2677)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2669..3478
                     /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(2736..2755)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3625..4213
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4114..4133
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4367..4384
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4543..4570)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4662..4748)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4797..4811,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4811,1..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 cggcaaagat ggtagagtcg tcaatttaaa atccaactat
      241 gccaacttta acattaaaat tgatggtcct ttttatatgt atgatgttgt tatggagcca
      301 aaaagtattc ggtccaaact gcaattattg atgacaaaaa gtgatgggca gccattttgt
      361 tttgatggtc gtagattatt tactacttct aaatggcatt caacggatga tgtagagaaa
      421 tttactgtta acaacgtcga aatttcaata aggttgctga caacatttgt aaaaagttct
      481 gaagaatatt atcaaatgat taatattgtg tttaataata ttcaaatttt tatgggacaa
      541 gagaaaattg ggcgacaatt ttttttgggt tcgaattgtg agagtggtgg aataaaagag
      601 ggtccgagtt ttttcaaaat ggagaatttt aaaattttgg gcggttattc aactaccatt
      661 accaggggta ctaatgaaga tggaacagct tcaactttat tatacttaga aagaataaat
      721 cgtgttctta atgaaaattc ggtaatcgct gtttatagaa gagatcaaat cgatcaatta
      781 atagggcgag acatcattac caaatacaat aataaaacat acagaatatc cgatattaaa
      841 gaaatgggta ttaatgacga aattgaattg aatggcacta aaatttcgta ttacaattac
      901 tttaaacagc gttataatat tactttgaaa aacgacaagc agccatttgt tatttctaga
      961 atcaaaagat caatgactca tcggcctaag gaaaaaagtg aaaatgaacc agaaggtcca
     1021 aaggaaacgg atcaaagttt atccattcca ggtgaattgt gttatttatg cggattttct
     1081 gatagtgaaa gaagcaatat aaatctacaa aaaagtttag gtggtgtgct aaaaagagag
     1141 cctcgtgaaa gattaaacga tataaaagat ttttgcaaat ggacaggtgg gggcaaatcc
     1201 aaagattata tgaaaacctg gaatatgact attgaggaac agcctattac gatacgaggt
     1261 cgtgaattac ctccggtaga tattatgtcc aacgatgtta aaattaatgc taaaactcca
     1321 gatgattgga aatttggtga agtaaaattt gatgttccta aaggtataac gcatagattt
     1381 ggcgttttag ttgtagatcg taatcctagt cattttaata attttatcga tgatgtaaaa
     1441 agagaaatta gtaggctcag aattaattac actattgata atatagcttc ttgtaggggt
     1501 aatgaagttg aagacgctct tactgaattt gttcgtgtta acaaagttca tttggcgtta
     1561 gtatttatac ctgatgataa ggtatatgca aaagtcaaaa actttactat gtctactgga
     1621 ttgttgacac aatgtgtgac tcaaagaaat ggaagtaacc gagatgatag acgccgtaag
     1681 accgtggctg ataaatctgt tatgcagata tttgctaaat taggatacga tccgtggggt
     1741 atcagtttaa agatggctcc aacaatgatt gtaggaatag atactttcca tagtaaaact
     1801 ggtaagagat cggtacaagc atccgtgttt tctataagtt ctaagttttc tcagtatatg
     1861 agttttgtca attcccctaa gggtaaaaat gaatttcatg agaacttggg caaaaatttc
     1921 ttaacggcaa ttactgcatt ccaacaaaaa tttggcacta tgcctcaacg atttattatc
     1981 tatcgtgatg gggtaggaga ttcccaatta ttcctcacga aaaaattcga gacagatgct
     2041 gtaatgaaaa tggtcgaaaa gatttatgag aatcaaaatt tgcctcaaat aatttatgta
     2101 gttgttaaaa aacggattag tgtaaaattc tttaaagatg gagctaatcc gaatccggga
     2161 acagtggttg atgaaaagat tgttaaacct aacttttatg aattctattt ggtttcacag
     2221 cgaactacta aaggaacagc atctccaacg aattataatg ttcttatgga tacgaagtat
     2281 tgtaataaga aaaccaacga agtgtcccca atggctccaa gcgttttgca acaaataact
     2341 tactctctta ctcacttgta ttttaattgg atgggtacaa taagagttcc tgtacccgtt
     2401 cattacgggt agaattggcc gggcccggta cccaattcgc cctatagtga gtcgtattac
     2461 ttccacataa ccccttgggg cctctaaacg ggtcttgagg ggttttttga ggatccccta
     2521 tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa
     2581 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     2641 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc
     2701 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     2761 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt
     2821 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     2881 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     2941 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata
     3001 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     3061 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     3121 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     3181 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt
     3241 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     3301 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     3361 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     3421 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     3481 agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc
     3541 aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     3601 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3661 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3721 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     3781 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3841 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3901 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3961 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4021 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4081 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4141 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4201 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4261 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     4321 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     4381 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     4441 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     4501 taatacgcgt accgctagcc aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg
     4561 atggccttct gcttagtttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct
     4621 ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga
     4681 gcgttcaccg acaaacaaca gataaaacga aaggcccagt cttccgactg agcctttcgt
     4741 tttatttgat gcctggcagt tccctactct cgcgttaacg ctagcatgga tgttttccca
     4801 gtcacgacgt t
//
