LOCUS       sequence-208802-        3796 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 3796)
  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 3796)
  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: Ddo_porcupine-1
FEATURES             Location/Qualifiers
     source          1..3796
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(59..145)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     194..216
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     208..225
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     209..225
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(232..279)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        284..302
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     360..376
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     360..376
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(1634..1652)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1710..1728)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1733..1749)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1733..1749)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1851..1870)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1862..2671
                     /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(1929..1948)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2818..3406
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3307..3326
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3560..3577
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3736..3763)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat
       61 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
      121 ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta
      181 gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gcaaaaaacc
      241 cctcaagacc cgtttagagg ccccaagggg ttatgctagg aagtaatacg actcactata
      301 gggagaccgg cagatctgat atcatcgatg aattcgagct ccaccgcggt ggcggccgct
      361 ctagaactag tggatccacc ggttccatgg ctagcggcat taccatcccg cgatgtcaga
      421 attctatggc ggatcaacga ttcttgcccc attgacgaat attgtcggca ttccggtcga
      481 tcaaataaat tttatcgttt gtagtttatt gtcaattggt ctcggacaat tgatgcgtca
      541 taaatttccg cctcataaat tatcagggaa tactcgaggg ttaattgaaa cgttttttgg
      601 aattttaatc gttctatttt gtttcggtta tcaggtaaaa tatttgttac tccaatcgac
      661 cgtctcgtat gtattattaa tggttttccc gtcaagaatt gcagcccatc ttgtcactta
      721 ctggtgttta gggttcatgt cagtcgttca tttataccga atgcactatg attacggtgg
      781 ttacacactc gacatttcgg gaccggtgat gattcaaaca cagaaactga cgagtcttgc
      841 atttaattta ctcgacggac atcgattaaa cgaaggacaa aaattacaac agaaaaacca
      901 cgaatcacac gccgttcgaa aactccccga cgccgttaag tttttcggtt atacgttcta
      961 tttccacgga atattagccg gtccgtttgt gttttataat gagtacaaag aatacattgt
     1021 tggttacgac aagaaatgga ttccatgcgc caagaaacaa ttgtcggcgg cgttatgttt
     1081 atcggtgatt cacggagttc tgacggcata tgtcgcccca caatttccat atattttcgt
     1141 ttgtagtacc gattatcaac gaatgcatat gacccgacgt attatgttta cgatgatttc
     1201 cttttttcta gtgagacaga aatactattt tgcctggtcg attgttgaag ccggtgcgtt
     1261 aagttccggt ttgggattcg gaggaaaaaa cgagaacgga gaaaatgatt ggtctaaggt
     1321 gaaaaattac gattttatgg caattgaacg cggtaatagc ttgaaagtat tagtcgacgg
     1381 ttggaacatt tcaacgacga gatggttgag agagttgatt tacgaacggg ctccgttgtt
     1441 ttgtcggacg attttagtgt ttgcgaccag tgcattttgg catggattct atcctggtta
     1501 ttatatgatg tttttaacgt tctttctctt tacgttttgc gctcggaccg ttcggaaaca
     1561 tattcgaccg ttgtttttag ttagcccgat caatcttccg ggtagaattg gccgggcccg
     1621 gtacccaatt cgccctatag tgagtcgtat tacttccaca taaccccttg gggcctctaa
     1681 acgggtcttg aggggttttt tgaggatccc ctatagtgag tcgtattaca tggtcatagc
     1741 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     1801 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     1861 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     1921 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     1981 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     2041 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     2101 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     2161 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     2221 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     2281 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     2341 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     2401 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     2461 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     2521 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     2581 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     2641 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     2701 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     2761 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2821 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2881 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2941 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     3001 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3061 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3121 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3181 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3241 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3301 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3361 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3421 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     3481 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     3541 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     3601 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     3661 cgactggaaa gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta gccaggaaga
     3721 gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttagt ttgatgcctg
     3781 gcagtttatg gcgggc
//
