LOCUS       sequence-208374-        3734 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 3734)
  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 3734)
  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: Gsp_porcupine-1
FEATURES             Location/Qualifiers
     source          1..3734
                     /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(1364..1382)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1440..1458)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1463..1479)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1463..1479)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1581..1600)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1592..2401
                     /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(1659..1678)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2548..3136
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3037..3056
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3290..3307
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3466..3493)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3585..3671)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3720..3734,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3734,1..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 cgatgttatg ggtatcccag tggatcaaat aaattttatt
      241 gtttgcagtt tattatcaat tggattaggt caattaatga gatacaaatt tcctccaaac
      301 aaagtctctg gaaatattag aggcctaatt gaaacattct ttggaatttt aatagtactt
      361 ttctgttacg gatatcaagt gaaatatttg ttgctacaat ctactgtttc gtatatttta
      421 ttactgctgc taccttcaaa aatatcagca catctagtaa cttactggtg catgggattt
      481 atggcagtgg ttcatttata cagaatgcat tacgattatg gtggttatac actcgatata
      541 tcaggtccag tgatgataca aacacaaaag ttaactagtc ttgcatttaa tcttcttgat
      601 ggacatcgac taaatcaagg aataaaatta attcagaaaa atcatgaatc tcatgctgtg
      661 aaaaagttac cagatgccat aaagtttttt ggttatacat tctattttca cggtatttta
      721 gcaggtccat ttgtattcta taacgaatat aaagaatata ttattggtta taatgacaaa
      781 catattcctt gtgcaaagaa gcaattatct gcagctttgt gtctatcagc aattcatggt
      841 ttattaacgg cgtacgtagc accacaattt ccatacatat ttgtgtgcag tactgattat
      901 caacgaatgc atatgactcg tcgaataatg tttacaatga tatcattttt tctcgttcgt
      961 caaaaatatt attttgcatg gtcaatggtt gaagctggag ctcttagtac tggtttagga
     1021 tattgtggga aggataaaaa tggggaaaat aattggtcta gagtaaaaaa ctatgatttt
     1081 atggcaatag aaagtggtag cagtttgaag atattagttg acggttggaa tatttctaca
     1141 acgagatggt tacgagaact gatttatgaa cgagctcctc tattttgcag aacaataatg
     1201 gtttttacag ttagtgcatt ttggcatggt ttttatcctg gttattacat gatgttttta
     1261 acattttctc tatttacttt ttgcgcaaga actgtaagaa aacatattcg acccctattt
     1321 ctagtcaacg ggtagaattg gccgggcccg gtacccaatt cgccctatag tgagtcgtat
     1381 tacttccaca taaccccttg gggcctctaa acgggtcttg aggggttttt tgaggatccc
     1441 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca
     1501 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
     1561 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag
     1621 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     1681 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt
     1741 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     1801 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     1861 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga
     1921 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     1981 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     2041 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     2101 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     2161 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     2221 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     2281 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     2341 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     2401 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg
     2461 cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     2521 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2581 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2641 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     2701 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     2761 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     2821 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     2881 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     2941 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3001 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3061 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3121 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3181 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     3241 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     3301 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     3361 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     3421 aattaatacg cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc
     3481 aggatggcct tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac
     3541 cctccgggcc gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg
     3601 agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt
     3661 cgttttattt gatgcctggc agttccctac tctcgcgtta acgctagcat ggatgttttc
     3721 ccagtcacga cgtt
//
