LOCUS       sequence-208225-        4123 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 4123)
  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 4123)
  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_PC2
FEATURES             Location/Qualifiers
     source          1..4123
                     /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(1753..1771)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1829..1847)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1852..1868)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1852..1868)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1970..1989)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1981..2790
                     /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(2048..2067)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2937..3525
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3426..3445
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3679..3696
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3855..3882)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3974..4060)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4109..4123,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4123,1..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 cgacaagcac gtattccact tccgacaata aataaaaaag
      241 gaattcctgg aagagaagct acagataaca ttaaaaatat gaatttacca actgatcctc
      301 tattttcaaa ggaatggtat attcggaaca ctggtcaagc agaaggagta aaagacttag
      361 atcttaacgt tctttcagca tggtctcaag gcattactgg taaaggtgtt actactgcaa
      421 ttatggatga cggtatagat tatcttcacc cggatctgtc taaaaattat aatgctgaaa
      481 gtagctatga tttcagcagt aacgatccat atccatttcc tcgttacacc gacgactggt
      541 ttaacagtca tggaacgcga tgtgcaggtg aaatttcagc tgagagagac aatggaatct
      601 gcggtgttgg ggttgcatat gattctaaga tcgccggatt acgaatgctt gaccaaccat
      661 atatgacaga tctaatcgag gcagctgcaa tgagctatga cagggataat attgatattt
      721 attctgcaag ttggggaccc acagatgacg gtagaacggt agatggaccc agaaatgaaa
      781 cgatgagagc aattgttgaa ggtgtaaata aaggtcggaa aggtaaagga agtatttatg
      841 tgtgggcttc cggagatgga ggagctgcag atgattgcaa ttgtgatggg tatgcagcga
      901 gcatgtggac aataagtatt aattcagcga ctaatgatgg agagacagca agttatgatg
      961 aatcatgttc atcaactctt gcatctacat tttccaatgg caaggcttgg tttagtcttg
     1021 atgcaggagt aggtacagta gatttatatg gacaatgtac acttcatcat tccggaacat
     1081 cagctgctgc tcctgaagct gctggagtgt ttgcattggc tttagaagct aacccagaaa
     1141 ttggatggcg agatatgcaa cacttaacag ttttaactag taaaagaaat cacttacatg
     1201 acagagaccg ccgacataat tggacaataa atggagcagg attggaattt aatcatctct
     1261 ttggttacgg agttcttgat gctggagata tggtccgagt agcaaaaaga tgggttaatg
     1321 tcccagaacg attccactgt atagcaggac aaatcgataa gcgctaccga tttaacgtca
     1381 gcagagcgct aatattagaa atcgatactg atgcctgtaa gtcagactcg gggcatcgag
     1441 tcaactattt ggaacatgtc caagcatttg tttcagtcaa agctactcga agaggagata
     1501 taacaatgtt tcttaccagt cctatgggta caacttcaat gattcttagt catcgaccaa
     1561 aagatgatga ttcttctcgt ggttttacta aatggccatt tatgacaacg catacgtggg
     1621 gagaaaatcc acgtggtaaa tggcgattaa cagtaatgtt tgaaacaaaa gagccactga
     1681 gtcaaggatt atttactgag tgggtccttg ttttacacgg gtagaattgg ccgggcccgg
     1741 tacccaattc gccctatagt gagtcgtatt acttccacat aaccccttgg ggcctctaaa
     1801 cgggtcttga ggggtttttt gaggatcccc tatagtgagt cgtattacat ggtcatagct
     1861 gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa
     1921 aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     1981 atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat
     2041 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc
     2101 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     2161 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     2221 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
     2281 ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     2341 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     2401 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     2461 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     2521 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     2581 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     2641 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     2701 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     2761 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg
     2821 cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg
     2881 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     2941 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3001 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3061 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     3121 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3181 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3241 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3301 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3361 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3421 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3481 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3541 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     3601 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     3661 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     3721 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     3781 gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag
     3841 tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg
     3901 cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat
     3961 ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
     4021 gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact
     4081 ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac gtt
//
