LOCUS       sequence-209332-        3186 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 3186)
  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 3186)
  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_sfrp-1
FEATURES             Location/Qualifiers
     source          1..3186
                     /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(816..834)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(892..910)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(915..931)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(915..931)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1033..1052)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1044..1853
                     /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(1111..1130)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2000..2588
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2489..2508
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2742..2759
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2918..2945)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3037..3123)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3172..3186,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3186,1..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 cgcatatctc ttgcttcctt ggcgattggc agcaatttca
      241 aaatggatat tcaactgaac aatgttttaa gattccagat aatttgacac tatgtctaaa
      301 tgttggttac aagaggatga ttttgcctaa ttatattcaa catgaatacc tggatgaagc
      361 cgttcagcat tcaaaggtgt ggattggttt agtaaattcc cattgccatg aagacataag
      421 gaaatttcta tgttctttat acgccccagt ctgcatcaat aatatgagaa ttcagaaagt
      481 acccccatgt aaagaactgt gtgtaagtgt gaaagaagca tgtttaccta gtatgaagtt
      541 atttggattt gaatggccaa atataatgaa atgttcttta ttcccaagtg taagaaattc
      601 gctttgtatc ccaagtactg ttagtaaagg aaaaaagtgt cgtccgtgcg tcaaatcctc
      661 atcatatgaa gttatggcta acaaattttg tctatcaaat atagtaattc gagccaaaat
      721 taaacgtatt gtaaaagaag gaaataatgg atttaaaatt gaattaggcc agaaaactgg
      781 cgggtagaat tggccgggcc cggtacccaa ttcgccctat agtgagtcgt attacttcca
      841 cataacccct tggggcctct aaacgggtct tgaggggttt tttgaggatc ccctatagtg
      901 agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct
      961 gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca
     1021 taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat
     1081 taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc
     1141 aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga
     1201 aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc
     1261 tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat
     1321 ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg
     1381 attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc
     1441 ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac
     1501 gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg
     1561 ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca
     1621 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta
     1681 ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact
     1741 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata
     1801 atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat
     1861 tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct
     1921 catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag
     1981 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     2041 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     2101 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     2161 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     2221 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     2281 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     2341 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     2401 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     2461 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     2521 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     2581 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     2641 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     2701 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     2761 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     2821 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata
     2881 cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc
     2941 cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg
     3001 ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt
     3061 caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat
     3121 ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac
     3181 gacgtt
//
