LOCUS       sequence-208838-        3244 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 3244)
  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 3244)
  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_slit-1
FEATURES             Location/Qualifiers
     source          1..3244
                     /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(874..892)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(950..968)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(973..989)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(973..989)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1091..1110)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1102..1911
                     /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(1169..1188)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2058..2646
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2547..2566
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2800..2817
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2976..3003)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3095..3181)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3230..3244,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3244,1..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 cggtcaagtg cgtaggtaga ggaataaaga ctgttcccca
      241 ggatttgccc accagtatca ttgaattaga tttacacgat aatgccatta atgttctgaa
      301 gcagaatgca ttatcaaaat ataagaattt gaaaaaaata ctgctaaata gtaatgaaat
      361 gatcgacatt gagccgggag catttgaagg tcttagacag ttgacttctt tgtacctaaa
      421 cggtaacaag ctcaagtcca ttcgcaaaaa tgctctcttg ggattatctt cactgatgtt
      481 tttatatatt cacaataacg atatcgaatg tctacctcgc gatacttttg ttggtctttc
      541 aaaattatat ttattacatt tatcggaaaa taaattgacg acactgaaaa aagagactct
      601 tgatcctctc acaaaacttc gatatttata tcttgtgcat aatcctctga attgcaactg
      661 taaattgagt tggttagccg agtttctacg atttaaggaa gcaagtggta gtcctgtggc
      721 ctcatgtctg ttgcccaaga atctccacgg ccaaccggtc agaggcgtgc aatcgtatca
      781 gtttaattgc ggtcaagatg aagaggccga gtcagagtca catgacgaag actgcatacg
      841 ggtagaattg gccgggcccg gtacccaatt cgccctatag tgagtcgtat tacttccaca
      901 taaccccttg gggcctctaa acgggtcttg aggggttttt tgaggatccc ctatagtgag
      961 tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga
     1021 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
     1081 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta
     1141 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     1201 tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa
     1261 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     1321 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     1381 ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat
     1441 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     1501 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     1561 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     1621 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
     1681 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     1741 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     1801 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     1861 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     1921 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca
     1981 tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa
     2041 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2101 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2161 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     2221 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2281 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2341 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2401 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2461 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2521 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2581 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2641 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2701 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     2761 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     2821 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     2881 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg
     2941 cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct
     3001 tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc
     3061 gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca
     3121 ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt
     3181 gatgcctggc agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga
     3241 cgtt
//
