LOCUS       sequence-208837-        3334 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 3334)
  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 3334)
  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_ifb
FEATURES             Location/Qualifiers
     source          1..3334
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(13..30)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(184..772)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(264..283)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(919..1728)
                     /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     1642..1661
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1720..1739
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1841..1857
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1841..1857
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1862..1880
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1938..1956
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2752..2768)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2752..2768)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(2826..2844)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2849..2896
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2903..2920)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2903..2919)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2912..2934)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      2983..3069
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3161..3188
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
       61 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
      121 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
      181 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      241 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      301 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      361 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      421 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      481 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      541 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      601 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
      661 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      721 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
      781 tttgatcttt tctacggggt ctgacgctca gtggaacgac gcgtaactca cgttaaggga
      841 ttttggtcat gagcttgcgc cgtcccgtca agtcagcgta atgctctgcc agtgttacaa
      901 ccaattaacc aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
      961 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     1021 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     1081 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     1141 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
     1201 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     1261 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
     1321 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     1381 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gtttttccgg ggatcgcagt
     1441 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     1501 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     1561 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaagc gatagattgt
     1621 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     1681 gttggaattt aatcgcggcc tcgacgtttc ccgttgaata tggctcataa caccccttgt
     1741 attactgttt atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc
     1801 aatgtaacat cagagatttt gagacacggg ccagagctgc caggaaacag ctatgaccat
     1861 gtaatacgac tcactatagg ggatcctcaa aaaacccctc aagacccgtt tagaggcccc
     1921 aaggggttat gtggaagtaa tacgactcac tatagggcga attgggtacc gggcccggcc
     1981 aattctaccc gctccgatgg atctttgtaa gccaatgaag tcaattcgcg aacctccgat
     2041 tcgtagatct tttttagaaa ttcgattttg tctcgtaagg attgcacctc attttgagcc
     2101 attatgtgat caagcgtttc tttgtcgagg tccttcttca attgggtaac ttcgtctttc
     2161 aatcgtccaa tctctttatc gtctcgtgtt ttttcgtctt ccaaacccat caatcgtcga
     2221 cgaagactag tcacttcatt ttcgtagtca ttaagctggg aattcattct gtctatagtt
     2281 tctttgtcag cattatgtat ttgattcact tcactaagct caatctctga ttcctgcagc
     2341 aaatcctcaa tagaattgat tctcatctcc agttgcacct ttgcaccttc agcagaatcc
     2401 aaagatttcc gcaagttatt catatcatta tcatactgat ccttgattcg ttcagtttct
     2461 ttgccccatt gattttttag gtccaccaat tcgtcggtca gggctctatt ttcatcctcg
     2521 agatgcttta ccttgtctat gtagtttgcg aatcgatcat tcaactcctg aagtgcttgt
     2581 cgttctttct cacgaccgtt cattaaggtg ctgacgcctg attgtacaat attagacgta
     2641 gcacctgaac tggcattgac gccgtagctt ttcttcattt cgattcgtcg aacattagta
     2701 acactttgac ctccgaccgg gatggtaatg ccgctagcca tggaaccggt ggatccacta
     2761 gttctagagc ggccgccacc gcggtggagc tcgaattcat cgatgatatc agatctgccg
     2821 gtctccctat agtgagtcgt attacttcct agcataaccc cttggggcct ctaaacgggt
     2881 cttgaggggt tttttgctta agactggccg tcgttttaca acgtcgtgac tgggaaaaca
     2941 tccatgctag cgttaacgcg agagtaggga actgccaggc atcaaataaa acgaaaggct
     3001 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
     3061 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
     3121 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
     3181 ggcctttttg cgtttctaca aactcttcct ggctagcggt acgcgtatta attgcgttgc
     3241 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     3301 aacgcgcggg gagaggcggt ttgcgtattg ggcg
//
