LOCUS       sequence-209285-        3460 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 3460)
  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 3460)
  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_sfrp-1
FEATURES             Location/Qualifiers
     source          1..3460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1..17
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        22..40
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        98..116
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1038..1054)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1038..1054)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(1112..1130)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1135..1182
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1189..1206)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1189..1205)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1198..1220)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      1269..1355
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1447..1474
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1633..1650)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1804..2392)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1884..1903)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2539..3348)
                     /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     3262..3281
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3340..3359
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 caggaaacag ctatgaccat gtaatacgac tcactatagg ggatcctcaa aaaacccctc
       61 aagacccgtt tagaggcccc aaggggttat gtggaagtaa tacgactcac tatagggcga
      121 attgggtacc gggcccggcc aattctaccc gtaagcatac ttgttcgtcc gtctgaattg
      181 tttggaagat tcaatcggca taattcgttt ccaaatttcc gtaaaacatt tatactgttc
      241 ctaaaatcta tattccttct tttccatttg gtgataaaat ttacaattgg ctgacctttg
      301 acaatattta ccattatcaa ccattttcct tttaaagctt tattttcaaa ttttaatttc
      361 gtgcaattac actgtagcca aatttgtaat gaactttgac tttgattaga ttgaaacttt
      421 aaaaatcgag atttttgact caaatcaatt atatatccat tgtctttctt tgctgtgatg
      481 cgtctgattt ttcctcttat aacgatattc gataaacaga atttgttagc catgacttca
      541 taagaagctg atttgacaca tgatcgacat tttttatctt tgtggattgt actgggaata
      601 cagagtgaat ttcttccact tggaaataaa gaacattcca taatattagg ccattcaaat
      661 ccaaacaatt tcatacttgg taagcatgat tctttcacac ttaaacataa ttctttacat
      721 ggaggcacct tttgaatttt catattatat atacaaacag gagcgtacaa agaacataaa
      781 aattttttta tatcttcatg gcagtttgaa ttcactaaac caatccatac ttttgaatgc
      841 tgaacagttt cttctaaata ttcgtgttga atataatttg gcaaaatcat tcttttatat
      901 ccaacattga gacaaagcgt taagttatct ggaattttgt aacattgttc tgcagaataa
      961 ccattttgaa attgctgcca gtcgcccagg aagcaagcga tttcgggatg gtaatgccgc
     1021 tagccatgga accggtggat ccactagttc tagagcggcc gccaccgcgg tggagctcga
     1081 attcatcgat gatatcagat ctgccggtct ccctatagtg agtcgtatta cttcctagca
     1141 taaccccttg gggcctctaa acgggtcttg aggggttttt tgcttaagac tggccgtcgt
     1201 tttacaacgt cgtgactggg aaaacatcca tgctagcgtt aacgcgagag tagggaactg
     1261 ccaggcatca aataaaacga aaggctcagt cggaagactg ggcctttcgt tttatctgtt
     1321 gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg
     1381 tgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa
     1441 ctaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact cttcctggct
     1501 agcggtacgc gtattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     1561 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     1621 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1681 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1741 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1801 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1861 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1921 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1981 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2041 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2101 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2161 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2221 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2281 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2341 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2401 tttgatcttt tctacggggt ctgacgctca gtggaacgac gcgtaactca cgttaaggga
     2461 ttttggtcat gagcttgcgc cgtcccgtca agtcagcgta atgctctgcc agtgttacaa
     2521 ccaattaacc aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
     2581 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     2641 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     2701 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     2761 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
     2821 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     2881 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
     2941 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     3001 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gtttttccgg ggatcgcagt
     3061 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     3121 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     3181 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaagc gatagattgt
     3241 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     3301 gttggaattt aatcgcggcc tcgacgtttc ccgttgaata tggctcataa caccccttgt
     3361 attactgttt atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc
     3421 aatgtaacat cagagatttt gagacacggg ccagagctgc
//
