LOCUS       sequence-208381-        4818 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 4818)
  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 4818)
  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: Gsp_beta-catenin-1
FEATURES             Location/Qualifiers
     source          1..4818
                     /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(2448..2466)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2524..2542)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2547..2563)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2547..2563)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2665..2684)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2676..3485
                     /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(2743..2762)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3632..4220
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4121..4140
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4374..4391
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4550..4577)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4669..4755)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4804..4818,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4818,1..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 cggctgtgag atttttagct ggagccatat ataatatgtc
      241 tcagaagaaa aatggcttaa agattatttt tgaagcaaat gtcataccgt gcttagtaaa
      301 attgcttgga ttttctatgg aatcagtttt attttacgca ataacaactt tacataattt
      361 attgttgtat caagatggtg ggaaagaagc agttcgatca agtggttgta ttcctaaaat
      421 ggtagcttta ttacaaaaaa ataacatcaa attcttaacg atatgtactg attgcttgca
      481 aattttagct tttaatcatc aaccaagtaa attagaaata ctgaagcatg gaggtccttt
      541 acatttaatc cacattatta aaagttatga ctatgaaaaa cttttgtgga ctgcaacaag
      601 agttttaaaa gttttaagtg tttgttctgc gaataaacct gttataataa gagaaggtgg
      661 aatggatgct ttaacaaaca tcttgtatca tacgatacaa aagaatacaa ccattgaatc
      721 ttttccgcca gtattaccaa cagaaaataa tgaatcatat ccacatttta gccagaggct
      781 tttacacaat tgcttatgga ctctaagaaa tttgtctgat gctgctacaa gattaaattt
      841 tgatcatttg cttaaagttt tggttcaaat tcttatgaat gcttttaata catttcaaag
      901 acaaagtact catatagata caaatgtgat aacttgtgct gcgggaatat tatcgaattt
      961 gacttgcaac aatcaatata ataaaattac atttttcaaa ttaggaggag ttgaagctat
     1021 tttgagaact attgaatgga atttagcatt gacaaatcct aatgtaccta ataaaataaa
     1081 tcagaataat tattttccag atgatatttt tgagccatgc atatgtactt taaggcatat
     1141 ttcaagtcgt catgaagaag ctgaagcagc ccaagcgtca atattgcatt ttcagggctt
     1201 acaaatttta attagaatca tcgaaatgca acttggacct gatctttttc atatgcaatc
     1261 acatcagcca ataatgccta atctttttct atcatattac aaaattaatt ggagtctcat
     1321 taaagcaatc attggtctca taagaaattt agctatgtct ccttgtaatt tcgtaccaat
     1381 tcgtgaaaag ggtttcggat ggccaatgat tgttttgatt aatcgagccc aaatagattt
     1441 gttaaataaa actgcttgtg ctagtatttc gcccaaagat aaattattag aagaaattat
     1501 agaagtgtct tgtggagctt tacatatgtt agctaaagat ccgggaattc gttcacaaat
     1561 tatttcaatg aagatttggc agcgattaat taactttcct caacagattg gcactcagtc
     1621 ttctgtgttt gtaattgcta gtttgttgtt taatcactta gaatcaattc aaagagtgac
     1681 tgctggactt ctagtagaag tttcagccga acgaatggga ttggaattaa ttattagtca
     1741 aatcgtgatt acgaataagc taagtgaaat ggttcattca aataatgagg caatatcaac
     1801 atatgcatcg gctattctaa tcagagcaac cgaagagaaa aataaaagat atacggcatc
     1861 tcataattac aatgttcctc agaatccaaa taatttgaga aataatttat ctatatcttt
     1921 gcctgctaat caagttagta atccaggata cttgaattct atattaaact cgcctattat
     1981 gcacaatcag ccatcttgtg taggtcacag acataattcg gttacacaat gtggtgatca
     2041 aaatatttct accaatgaaa catgtcaaat taatgacgta gatgtttcag attatcatat
     2101 gaccttttct aatactctac agccacaaaa tagatattca tacgcctcaa attacttgga
     2161 tccaaattat agtcttctaa atcaaactgt aaatccggaa atctcgatga gacaacattt
     2221 taagcaactt ggtcccatta atatttcaaa tgaatgtatt aaccaagaca tacaaaaaac
     2281 atttacaaat ccaagtttca agaatcctac atctcatcaa atgccactta ctggtaattt
     2341 gtgtccatca aatgacaact ctaaaaagga aatgatggat gtatgtatgt tagaagcaga
     2401 ctggattgaa accgggtaga attggccggg cccggtaccc aattcgccct atagtgagtc
     2461 gtattacttc cacataaccc cttggggcct ctaaacgggt cttgaggggt tttttgagga
     2521 tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt
     2581 ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac
     2641 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt
     2701 cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
     2761 ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag
     2821 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
     2881 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
     2941 ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc caggtattag
     3001 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
     3061 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc
     3121 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
     3181 atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg
     3241 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
     3301 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
     3361 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     3421 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
     3481 tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg acttgacggg
     3541 acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt
     3601 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3661 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3721 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     3781 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3841 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3901 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3961 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4021 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4081 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4141 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4201 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4261 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     4321 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     4381 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     4441 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     4501 acgcaattaa tacgcgtacc gctagccagg aagagtttgt agaaacgcaa aaaggccatc
     4561 cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc gtcctgcccg
     4621 ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat ttgtcctact
     4681 caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt ccgactgagc
     4741 ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta gcatggatgt
     4801 tttcccagtc acgacgtt
//
