LOCUS       sequence-208829-        3422 bp    DNA     circular SYN 14-APR-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 3422)
  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 3422)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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_sfrp-1
FEATURES             Location/Qualifiers
     source          1..3422
                     /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(1052..1070)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1128..1146)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1151..1167)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1151..1167)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1269..1288)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1280..2089
                     /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(1347..1366)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2236..2824
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2725..2744
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2978..2995
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3154..3181)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3273..3359)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3408..3422,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3422,1..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 cgggttatac gacggatcaa tgttacaaag ttcccgaaaa
      241 cttttcaatg tgtcataatg ttggttataa caaaatggtt ctacctaatt ttatacaaca
      301 cgagtatatg gaagaagcga tagctcattc tcgagtatgg ttaagtctag ttaagtctca
      361 atgtcatctt gatgtgagaa agtttttgtg ttctttatat gctcctgtat gtattacaaa
      421 taaaagaata caaaaagttc ccccatgtcg agaactatgc ataattgtca aggaagcttg
      481 tttaccaagt atgaagatgt acggctttaa ctggccggag attatgaaat gttccaaatt
      541 tccaccagta caaaatgcat tatgtatacc gaaaacaaca gtgaaaaaaa aaaattgtga
      601 attatgtgtt aaaaatgcgc cttacgaagt gatagcaaat agattttgtt tatctcccat
      661 agtaataagg gctaaaatca aacgaattat tcccaaagaa tcggggtttc aaatagtttt
      721 gcatcaaaaa tccaaatttt taaaatttca atcaaattct agtcgggatt ccgtcaattt
      781 atggctgaaa tgtaattgtt tgaatctgaa atttcagcaa agagcccttc gaggaaggtg
      841 gatcatcatg gctcgaatcg tcgaagatag atttgttgtg aatttcattt ccaaatggaa
      901 aaaaaagaaa gaggaattta aaaattctat tagaattatt cagaaattcg gaaatgttgt
      961 ttgtaaatcg aagcttaagg atggcataag tctccaggaa cgatatgtgg cctatacggg
     1021 tagaattggc cgggcccggt acccaattcg ccctatagtg agtcgtatta cttccacata
     1081 accccttggg gcctctaaac gggtcttgag gggttttttg aggatcccct atagtgagtc
     1141 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     1201 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     1261 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     1321 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     1381 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     1441 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     1501 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     1561 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     1621 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     1681 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     1741 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     1801 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     1861 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     1921 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     1981 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     2041 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     2101 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     2161 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     2221 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2281 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2341 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     2401 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2461 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2521 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2581 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2641 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2701 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2761 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2821 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2881 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2941 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     3001 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     3061 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg
     3121 taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc
     3181 tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt
     3241 tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc
     3301 gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga
     3361 tgcctggcag ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg
     3421 tt
//
