LOCUS       sequence-208831-        5277 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RNAi studies in planarians.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5277)
  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 5277)
  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_beta-catenin-1
FEATURES             Location/Qualifiers
     source          1..5277
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(137..164)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(256..342)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     391..413
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     405..422
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     406..422
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(429..476)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        481..499
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     557..573
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     557..573
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(3312..3330)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3388..3406)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3411..3427)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3411..3427)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3529..3548)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3540..4349
                     /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(3607..3626)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4496..5084
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4985..5004
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5238..5255
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
       61 ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag
      121 agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct
      181 ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa
      241 atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa
      301 acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta
      361 ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa acgacggcca
      421 gtcttaagca aaaaacccct caagacccgt ttagaggccc caaggggtta tgctaggaag
      481 taatacgact cactataggg agaccggcag atctgatatc atcgatgaat tcgagctcca
      541 ccgcggtggc ggccgctcta gaactagtgg atccaccggt tccatggcta gcggcattac
      601 catcccgttg gaaagtcaca gtcacagtgt cagtagtaaa catggttacg atgaaatgga
      661 atcagaagat caaaataaaa attcaaaaga atggaattat aatacatcaa acaataacgt
      721 taatgatgga ttgtcgaatg acatttgcat gctgactcca gataccccag gaagttctgg
      781 atgtcttaga cacgggggtg aaggtatggc cgacgtcaac ttgtcgactt gttacgatgg
      841 taaagaagaa aacaatagaa atccagattt tgcaatagac atcgataaaa ttgaagctga
      901 aagtgccata cctgatctta tcaaactgat tagcgatgaa gacgatgctg tagttataaa
      961 cgaggctaca atgatggttt ttcagttgag taaaactgat gcaattgatg caataataaa
     1021 ttcaaaagaa atgatatcat gtatgattga agcaatgaat aaagccgaac acgcagaagc
     1081 agttagattt ttagctggtg ctatttataa tatgtctcaa aagaaaaacg gactaaaaat
     1141 tattttcgaa gccaatgtca taccgtgttt ggttaaattg ttgggatttt caatggaatc
     1201 tgtgttattt tatgcaatca ccaccttaca caatttacta ttatatcaag aaggcggcaa
     1261 agaagctgtg agaattagtg gctgcattcc taaaatggtt gctcttttgc agaaaaataa
     1321 catcaagttt ttgacgattt gtactgattg tttacaaatt ttggctttca atcatcagcc
     1381 gagtaaatta gagattttaa aacacggtgg tcctttgcat ttaattcata ttattaaaag
     1441 ttatgattat gaaaaacttt tatggacggc aacaagagtt ctgaaagttt taagtgtttg
     1501 ttcagctaat aagccagtta ttatacggga agggggaatg gatgcattga ccaatatttt
     1561 gtatcataca atacaaaaaa acacaaacat tgaatcctat ccacaagttg taccaaatga
     1621 aaacaatgaa tcatatccac actttagcca gagacttttg cacaactgtt tatggacact
     1681 tagaaattta tcggatgcag ctactcgtct aaatttcgat aatttactta aagttttggt
     1741 tcaaatttta gtgaatgctg ctaattcatt tcaaagacaa gctcctcata ttgatacaaa
     1801 tgttattaca tgtgcagctg gaattttatc taatttaact tgcaacaatc aatataataa
     1861 aataactttt tataaacttg gtggagtgga agctatttta agaactatcg aatggaattt
     1921 agcattgaca aatccaacta atgttccgaa caaattaacc caaaacaatt attttcctga
     1981 tgacattttt gagccatgta tatgtacttt aagacatatt tccagtcgcc atgaagatgc
     2041 tgaggctgcc caagctgcaa tattacattg tcaaggatta catattctca tcaagatcat
     2101 agaaatgcaa ctcggtccgg atctttttca tatgcaatct catcaaccaa ttatgcctaa
     2161 tctatttctt tcgtactata aaataaattg gagtctcata aaagcaatta tcggccttat
     2221 aagaaatctt gcaatgtttt catgcaattt cgtacctata cgagaacgag gtttttgttg
     2281 gccaatgatc gttttaatta atcgagctca aattgattta ttgaataaaa atgcatgttc
     2341 ttctatttcg ccgaaagata aattgctaga agaaatcatt gaagtgtctt gtggagcatt
     2401 acatatgctt gctaaagatc ctggtattcg agcacaaata acgtctatga agatatggca
     2461 aaggttaatc agttatccac aacaaattgg tattcaacca gctgtatatg taatttccag
     2521 ccttttgttt aaccatttag aatcaattca aagagtagca gctggtcttt tggttgaagt
     2581 ttcagctgag cgtttaggat tagaactaat tattaaccaa ttggtgatta ccaataaact
     2641 taatgaaatg gttcattcta ataatgaagc aatatcaaca tacgcttccg ctattctaat
     2701 tagagcaact gaagagaaaa gtaaaagata tgcttcatct cacagctata atgtcgttcc
     2761 acaaaatcca aacaatttga gaaataatct ttcgatttct ttaccgaata atcaaaatac
     2821 caaccctggt tatttgagct ctatattaaa ttctccgatt ttacacaatc agtcgccatg
     2881 tataagtcac agacataata ctgtagcaca atgtggagat caaactgttt caaatgaacc
     2941 aatatgccag ttaaatgatg tagaagtacc tgattatcat atgaattatt ctaatactct
     3001 tcaaccacaa aatcgatatc catataattc taactacatt gatccaaatt acagtctaat
     3061 caatcaaccg ggaaattcag atttaacaat gagacatttt aagcaactcg gtccaatcaa
     3121 tatttcgaat gaatgtgtta atcaagacgt tcaaaagaac ttttcgaaca caagttttaa
     3181 aagtccctca tctcattctt tatctcttag tggtaactta tgctcgtcta gtgacaattc
     3241 taagaaggaa ataatggacg tgtgcatgtt agacaccggg tagaattggc cgggcccggt
     3301 acccaattcg ccctatagtg agtcgtatta cttccacata accccttggg gcctctaaac
     3361 gggtcttgag gggttttttg aggatcccct atagtgagtc gtattacatg gtcatagctg
     3421 tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa
     3481 atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta
     3541 tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt
     3601 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct
     3661 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
     3721 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
     3781 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg
     3841 gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
     3901 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
     3961 gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg
     4021 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
     4081 ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
     4141 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
     4201 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
     4261 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
     4321 ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg taacactggc
     4381 agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga
     4441 gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4501 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4561 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4621 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     4681 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4741 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4801 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4861 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     4921 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4981 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     5041 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     5101 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     5161 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     5221 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatac
//
