LOCUS       sequence-209317-        5653 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 5653)
  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 5653)
  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: Pmo_beta-catenin-1
FEATURES             Location/Qualifiers
     source          1..5653
                     /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"
     CDS             complement(3229..3240)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(3283..3301)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3359..3377)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3382..3398)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3382..3398)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3500..3519)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3511..4320
                     /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(3578..3597)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4467..5055
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4956..4975
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5209..5226
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5385..5412)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5504..5590)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(5639..5653,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(5653,1..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 cggcctatcg aacataagca tcaatgataa ttttccaaat
      241 gatatttgta tgctaacacc agattctccc ggaagtaaca tctgtggcag acaaggtgga
      301 gaggcaatgg cggatgtcaa tttaacttct ggatatgatg gtggtaaaat agactgtgaa
      361 aaggagttcg gcttagatat agataagata gaagcagaag gtgctatacc agatttggtt
      421 aaactaatta aggatgaaga tgatgcagtg gttatcaatg aagctacaat gatggtattt
      481 caattaagta aatcagatgc tatcgatgct ttaattaatt caaatgaaat gatttcgtgt
      541 attattgaag ctatgaataa agctgaacat gcagatgcca ttagattctt agcgggcgct
      601 atttataaca tgtctcaaaa gaaatatgga ctgaaggtaa tttttgaggc aaatgtaata
      661 ccatgcttgg tgaaacttct tggattttca ttggaatccg ttttgtttta tgcaataaca
      721 acgttgcaca atttattact gtatcaagaa ggaggaaaag aagcagttag atcaagtagt
      781 tgtattccaa agcttgttac gcttctgtta aaaaataaca caaaattttt aacaatttgt
      841 accgactgct tacaaatttt ggcatttaat catcagccta gcaaaattga gatattaaag
      901 aatggtggac caatgcatct gattcatatt ataaaaaact ataactatga aaagttatta
      961 tggacagcta ctcgagtttt aaaggtcctc agtgtttgct cggcaaataa accagtcata
     1021 atcagagagg gaggaattga agcactcaca aacattttat atcacacaat tcaaagtaat
     1081 tcatcaaccg caccgtatca agctgaaccg cccaatgaaa ataatgaaac gcatcctcaa
     1141 tttagtcaga gactattgca taattgtctt tggactttac gaaatttatc ggatgcagca
     1201 actcggatta attttgatca tttattaaaa gttttagttc aaattctgat gagttcgtat
     1261 aattcgtttc aaagacaaaa tattcatatc gatactaacg ctgttacatg tgcagctgga
     1321 atattgtcta atttgacttg caataatcag ttcaataaaa ttgctttatt taagcttggt
     1381 ggagttgaag cgttaattaa agttatcgaa tggaattttg cttctctcaa taatccaaat
     1441 gcacaaaata aaaacaactc caacaatgcc ttctcggagg atatattaga accgtgcatt
     1501 tgtgcattgc gtcatttgac gagcagacat gaggaagcag aggctgttca gacggcactt
     1561 gttcaactgc aaggcttgtc cttgctgatc cgattattgg aaatgcaact cggacaggat
     1621 ctgtactata tgcaaactca gcaaccagtc atgcctaatc tgttcatgtc gtacaaaatt
     1681 aattggagtc tcataaaagc gatcataggt ctcatacgga atatggcaat gtcatcgtgc
     1741 aattttgcgt cgatacgaga acgtggtttt gtgtggccaa tgattgttct aattaatcgg
     1801 gcacaaatcg atgttgctaa caagaatccg ggtacatccg cggtgtcatc gaaagataaa
     1861 ttgctcgaag aaattatgga agtcacatgt ggtgcattgc acatgctcgc taaggatccc
     1921 ggcatccggg ctcaaattat atcaattaaa ttatggcagc gtttaattta ctgtccgcaa
     1981 caaattggaa ttcaatctgc catttacgtg atcgcaagtc ttctgttcaa tcatcttgaa
     2041 tcaattcaac gggtcgcagt tggacttctt gttgaactgt ctgcggaacg gatggccttg
     2101 gaaatgataa tgggacatat ggtaatcacc aacaaactca acgaaatggt gcactcgaaa
     2161 agcgaagcgg tggcaactta cgcatcggcc atattgatcc gagccacaga agaaaaaaat
     2221 aaacgttaca caccgtgtca caattacaat gtgccgcttc ctaataccgg atgctcgaga
     2281 ccgaatctct ccatctctct ctcatctaat cagtcccata attcgcctgg attccttaac
     2341 tcgatactgc attcgccggt catgcacaat cagccgcctt gtgttagtca tcggcacata
     2401 ccgtcccaat gcggggatca gaattacata aacaacgaac tattgggtca gatcggcgac
     2461 atggacgtat cagacatgca gatgaactac ggccagtcca cagggccccc gcccccgcag
     2521 acccagaaca ggtacccgta caaccccggc tacatggata ataacaatgg ctgcaccgtg
     2581 ctcctcgccg gcaatcagcc tccaccgccc caacagcagc agcagcagtc aatgtcatcg
     2641 gacatgagat acttcaagca gccaccgggc atcgtcaacg aatgtcacgt gaccaccgat
     2701 caccagaagg ccgcatacat aggcaagcct cgtgccccgc atcttcagca tccgtcctcg
     2761 atggacctcg gcggcactgg cgccaagatc aaagagccaa tggaactgtg catgccagac
     2821 gccgttgatt gggcggagtc ctcctgtggc caggacttcc cccaccacca ggggcactcg
     2881 tcccacaaat cctgactgtt caccaccgca taaaaagtat gttgactttt acttttgttt
     2941 atttattaga cagtttcgtt atcgttatta attaattaat taattactaa ttactatttc
     3001 gtgatgtaat tgtgatcatt tcgaatgaat gacggatgga caatgacttt gtagtggttg
     3061 tgattgcggc tgtattgatt cattgcgatg atggcgcctc tcacttgcat gcatgcacgc
     3121 acgcacgttt ctctctctct ctctctctcc tgaacgtcca gcgatacata cgtgcacaca
     3181 cccacacaca tgacacactg actgactgac tcacggtact tgctgctctc gaccttcgat
     3241 tgactaccgg gtagaattgg ccgggcccgg tacccaattc gccctatagt gagtcgtatt
     3301 acttccacat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gaggatcccc
     3361 tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa
     3421 aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct
     3481 gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcgagg
     3541 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
     3601 gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg
     3661 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
     3721 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
     3781 gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt attagaagaa
     3841 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
     3901 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg
     3961 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
     4021 tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca
     4081 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     4141 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     4201 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     4261 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     4321 tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc
     4381 gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac
     4441 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4501 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4561 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     4621 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4681 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4741 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4801 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4861 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4921 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     4981 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5041 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5101 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     5161 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     5221 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     5281 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     5341 attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca
     5401 ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc
     5461 ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga
     5521 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc
     5581 gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg gatgttttcc
     5641 cagtcacgac gtt
//
