LOCUS       sequence-209323-        3786 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 3786)
  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 3786)
  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_porcupine-1
FEATURES             Location/Qualifiers
     source          1..3786
                     /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(1416..1434)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1492..1510)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1515..1531)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1515..1531)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1633..1652)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1644..2453
                     /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(1711..1730)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2600..3188
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3089..3108
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3342..3359
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3518..3545)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3637..3723)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3772..3786,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3786,1..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 cgtattcttc cagcaggctt agccgctcat cttgtcacat
      241 tttggagctt aggttttatg ggagtggtac atatgtatcg catacatttt gattatggtg
      301 gctatacaat ggacatttca ggcccagtaa tgattcaaac tcagaagttg acaagtcttg
      361 ctttcaattt acttgatgga tggagactta atcacggcta tactttacct caaaagaagc
      421 acgaaactca tgcagtaaag gaacttccaa atgctgttaa ttattttagt tatcttatgt
      481 atttccatgg tatacaggca ggacctttta cattttacaa cgagtacaga gaatacctaa
      541 caggatatga cattaaagca ataccatgtg ccaaaaagca aataagagca gctttatgct
      601 tatcgattat tcatggttta cttacagcat atgttgcacc tcaattccct tatattttta
      661 tatacagtac agactatcaa cgaatgcata tgagtcgccg aatcgtattt acaatgattt
      721 cttttttcct tgtgcgtcaa aaatattatt ttgcttggtc tatttctgaa gcagcatcac
      781 ttagtgcggg tcttggctac agtggcaaga ctaaaaacat ggaaaacgac tggtcacgta
      841 ttaagaatta tgattttatg gctattgaaa gtggtagttg tttgaaagta attggagatg
      901 gatggaatat gtcaacaact aggtggcttc gagaactcgt atatgagaga tcacctctat
      961 tctgtcgaac attattagta tttgcaacaa gtgcattctg gcatggattc tatcccggtt
     1021 attacttaat gtttggaacc tttgcaattt ttaccttctg cgcaagaact gtacgtaagc
     1081 atgttagacc tttatttatg actaatccag taacatttca actatatcac gtaataacaa
     1141 aaattataac taatttagca cttaattatg gtcaggcgcc attttattta ttagaattcc
     1201 aaactgcact cttattttgg aaaccgtatt atttttttgg acacgtcatt gcaatatcta
     1261 taatattaat gttccaagtt cgaccaatta tcaatgataa agtaggacaa attaatttca
     1321 atctcacaaa gaaagctaac ttaaattttt tacttctaaa ggaacgtctg ctgttaaaac
     1381 cgggtagaat tggccgggcc cggtacccaa ttcgccctat agtgagtcgt attacttcca
     1441 cataacccct tggggcctct aaacgggtct tgaggggttt tttgaggatc ccctatagtg
     1501 agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct
     1561 gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca
     1621 taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat
     1681 taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc
     1741 aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga
     1801 aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc
     1861 tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat
     1921 ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg
     1981 attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc
     2041 ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac
     2101 gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg
     2161 ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca
     2221 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta
     2281 ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact
     2341 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata
     2401 atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat
     2461 tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct
     2521 catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag
     2581 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     2641 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     2701 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     2761 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     2821 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     2881 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     2941 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     3001 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3061 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     3121 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3181 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     3241 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     3301 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     3361 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     3421 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata
     3481 cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc
     3541 cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg
     3601 ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt
     3661 caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat
     3721 ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac
     3781 gacgtt
//
