LOCUS       sequence-208833-        3696 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 3696)
  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 3696)
  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_PC2
FEATURES             Location/Qualifiers
     source          1..3696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(35..52)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(206..794)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(286..305)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(941..1750)
                     /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     1664..1683
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1742..1761
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1863..1879
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1863..1879
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1884..1902
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1960..1978
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3136..3152)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3136..3152)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(3210..3228)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      3233..3280
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3287..3304)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3287..3303)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3296..3318)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      3367..3453
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3545..3572
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
       61 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
      121 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
      181 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
      241 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
      301 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
      361 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
      421 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
      481 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
      541 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
      601 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
      661 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
      721 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
      781 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
      841 acgcgtaact cacgttaagg gattttggtc atgagcttgc gccgtcccgt caagtcagcg
      901 taatgctctg ccagtgttac aaccaattaa ccaattctga ttagaaaaac tcatcgagca
      961 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
     1021 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
     1081 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
     1141 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
     1201 aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
     1261 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata
     1321 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
     1381 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
     1441 ctgtttttcc ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
     1501 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
     1561 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
     1621 tcccatacaa gcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
     1681 acccatataa atcagcatcc atgttggaat ttaatcgcgg cctcgacgtt tcccgttgaa
     1741 tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg
     1801 atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacacg ggccagagct
     1861 gccaggaaac agctatgacc atgtaatacg actcactata ggggatcctc aaaaaacccc
     1921 tcaagacccg tttagaggcc ccaaggggtt atgtggaagt aatacgactc actatagggc
     1981 gaattgggta ccgggcccgg ccaattctac ccgatcgtat ggtcgatctt gagtgccatg
     2041 aagaacgagg acccactcag agagcagtcc ttggatcagg gcgtcctctg tatcgaacat
     2101 gacggttagt cgccacttgc cgcgagggtt ctctccccag gtgtgcgtgg tcatgaaggg
     2161 ccatttggta aacccgcgga tcgagtcact gtctttctct cgacgactaa ggatcatcga
     2221 cgtcgttccc attggactcg tcagaaacat cgttatgtca cctcgacgac ttgctttcac
     2281 agtcacaaat gcttgtacgt gttctaagta gttcactctg tggttatcat cggatttgca
     2341 agcgtccgtg tcaatctcca gcacaagtgc tttgctgata ttaaacctcg atttttggtt
     2401 gatctctcca gcaatgcaat gaaacctttc tggaacgttg gcccacatct tcgccactcg
     2461 gaccatgtca cctgcgtcta gcaccccata gccgaataag tgattgaact cgagacctgc
     2521 accattaatc gtccagttat gtcgcccgtc cctgtcgtgc aaatggttcc tcttgctagt
     2581 cagtactgac aaatgctgca tgtctctcca gcccatttcc ccgttcgctt caagggcaag
     2641 agcaaagacc gctgctgcct ctggagcggc ggcggatgtg ccagaatggt ggagtgtaca
     2701 ttggccgtac agatccactg tgccgactcc ggcatccaga ctgaaccaag ccttgccatt
     2761 ggagaatgtg gaagcgaggg tcgaagaaca cgactcatca taactcgctg tttcgccgtc
     2821 attcgtagcg gaattgatgc tgatcgtcca catgctcgcc gcgtatccat cgcaattgca
     2881 atcgtctgag gcaccaccgt ctcctgaagc ccagacataa atgttacctt tgccgttgcg
     2941 gcccttattg accccgtcaa caatggccct catggtttcg ttgcggggtc catcgacagt
     3001 tctaccgtca tcggtggggc cccaactggc ggagtaaata tggatattgt cgcgatcatg
     3061 gctcatcgct gcagcttcaa tgagatccgt catgtatggt tcgggatggt aatgccgcta
     3121 gccatggaac cggtggatcc actagttcta gagcggccgc caccgcggtg gagctcgaat
     3181 tcatcgatga tatcagatct gccggtctcc ctatagtgag tcgtattact tcctagcata
     3241 accccttggg gcctctaaac gggtcttgag gggttttttg cttaagactg gccgtcgttt
     3301 tacaacgtcg tgactgggaa aacatccatg ctagcgttaa cgcgagagta gggaactgcc
     3361 aggcatcaaa taaaacgaaa ggctcagtcg gaagactggg cctttcgttt tatctgttgt
     3421 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgtg
     3481 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaact
     3541 aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct tcctggctag
     3601 cggtacgcgt attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     3661 gccagctgca ttaatgaatc ggccaacgcg cgggga
//
