LOCUS       sequence-209320-        4743 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 4743)
  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 4743)
  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: Pgr_piwi-1
FEATURES             Location/Qualifiers
     source          1..4743
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(6..23)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(177..765)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(257..276)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(912..1721)
                     /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     1635..1654
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1713..1732
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1834..1850
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1834..1850
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1855..1873
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1931..1949
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4154..4170)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4154..4170)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(4228..4246)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      4251..4298
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4305..4322)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4305..4321)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4314..4336)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      4385..4471
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4563..4590
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
       61 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
      121 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      181 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      241 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      301 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      361 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      421 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      481 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      541 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
      601 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
      661 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
      721 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
      781 ttttctacgg ggtctgacgc tcagtggaac gacgcgtaac tcacgttaag ggattttggt
      841 catgagcttg cgccgtcccg tcaagtcagc gtaatgctct gccagtgtta caaccaatta
      901 accaattctg attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
      961 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
     1021 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca
     1081 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
     1141 tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt
     1201 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
     1261 attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa
     1321 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct
     1381 gaatcaggat attcttctaa tacctggaat gctgtttttc cggggatcgc agtggtgagt
     1441 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc
     1501 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca
     1561 tgtttcagaa acaactctgg cgcatcgggc ttcccataca agcgatagat tgtcgcacct
     1621 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa
     1681 tttaatcgcg gcctcgacgt ttcccgttga atatggctca taacacccct tgtattactg
     1741 tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa
     1801 catcagagat tttgagacac gggccagagc tgccaggaaa cagctatgac catgtaatac
     1861 gactcactat aggggatcct caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt
     1921 tatgtggaag taatacgact cactataggg cgaattgggt accgggcccg gccaattcta
     1981 cccggtacct ttagtggttc gctgtgaaac aagatagaat tcatagaaat ttggtttaac
     2041 aactttttca tcaacaattg tgcctggatt tggattgtta ccgtctttaa aaaatttaac
     2101 actaattctt tttttaacga ttatgtatat aatttgtgga aggttttgat tttcataaac
     2161 agattcaatg attttcaaaa ccgcatccgt ttcgtacttc tttgtaaaat acaattgaga
     2221 atcaccaact ccatcacggt aaattataaa ccgctgaggc aaggtattaa atttatctct
     2281 aaataaattt attgcggcct tgaaattttg actcaaatta tcgtgatatt cacttttccc
     2341 tttcgatgaa ttaacaaaac ttaaatattg tgtaaattta gagcttatac taaatacaga
     2401 ggcttgtata gaccgcttgc cagttttact atgaaacgta tccaatcctg caaccatagt
     2461 tggtaccatt tttaattcaa ctccccatgg atcatacccc aacttagcaa acatttgcat
     2521 tacactttta tccgcaacta cttttctcct tcgatcatcg cgatttgtgc cattccgaac
     2581 agttacacat tgtgtcaaca aaccagtact tatggtaaaa tttttaactc ttgcatatat
     2641 tttatcatct ccaataaata caagagccaa atgaacctta tcacgtatga aatctgtaag
     2701 gaaatcttcg atttcattcg ttctgcatga caaaacagat ccaattgtgt gattgactct
     2761 taatctactc atttctcttt taacatcgtc aatgaattga ttgaactgcc ctggatttct
     2821 ttcaacgact aatattccaa atctatgttg tgtaccagcg gggatatcaa atttaacttc
     2881 cccaaatttc caatcgtccg atgatttcgt attaatacgt tgcttgtttg aaaacacctc
     2941 aacaccattg agttctctac ccatgattgt aacaggttga ttttctaatt ccaatcccca
     3001 tttttccata aaaggagatg cctttccttc ctttgtccag ttacaaaaat cactaatatc
     3061 tcgcaaacgt tcacgcggtt ctcgttttaa aacggaacct aagcttttct gtaaattaat
     3121 attacttctt tcactgtccg aaaagccaca caagaaacat aactcccctg gaatagacaa
     3181 gctttggtcg gcttctttgg gggcttcagg atcattttct agattttcct tttggataat
     3241 tccaattctt ctcattctag aaataacgaa tggctgctta ccggtggtta aggtaatatt
     3301 ataacgttgt ttaaagtagt caaaataact aattttaaca ccatttaatt caatttcttc
     3361 agatacgtca agttctttaa tatcgctaat tcgatatgtc ttgttgttat atttcgttat
     3421 gacatcacgg ccaattagca agtcaagttg attcttatta taaatgttat aaaccgaatt
     3481 ttcattcaag actcgattta ttcgttccaa ataaagtagt gtacaagcag taccgtcttg
     3541 atttgttcct cttgtaagag tcgtacaaaa tccacctaat attttaaaat tatctctctt
     3601 gaaaaatgtg ggtccacatt tagacaagct ctcattgcaa tttgatccaa gaaaaaaagc
     3661 tctaccaata cgctcttgtc ccataaaaac ttgaatatta ttaaagatta tattaatcat
     3721 ttgataatat tcttctgtat tttctataaa agtagccaaa aatctaattg atacctcttc
     3781 gtctttaatg ttaaacgttt caatgtcttc tttatgccat tgtgagggtg taaaaaggcg
     3841 acggccatca aaacaaaaag gttgattatc agcctttgtg gacgccacac tgtataataa
     3901 ttccaactta ccccgtagac ttttcattgg catttttacg tcatacataa aaaaaggacc
     3961 ttttggtccc actttaaaat ttgcataatt agattttaat gcaactgcac gaccatcttt
     4021 tccaacctta tcaacaatta attctggttg aagactcggt tcaatgacat aaaatctctt
     4081 ttgtaatcct cctcgtttgg ctacctttct ctcctccatc gggatggtaa tgccgctagc
     4141 catggaaccg gtggatccac tagttctaga gcggccgcca ccgcggtgga gctcgaattc
     4201 atcgatgata tcagatctgc cggtctccct atagtgagtc gtattacttc ctagcataac
     4261 cccttggggc ctctaaacgg gtcttgaggg gttttttgct taagactggc cgtcgtttta
     4321 caacgtcgtg actgggaaaa catccatgct agcgttaacg cgagagtagg gaactgccag
     4381 gcatcaaata aaacgaaagg ctcagtcgga agactgggcc tttcgtttta tctgttgttt
     4441 gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgtgaa
     4501 gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaactaa
     4561 gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttc ctggctagcg
     4621 gtacgcgtat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4681 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4741 tcc
//
