LOCUS       sequence-208803-        4020 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 4020)
  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 4020)
  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_ifb
FEATURES             Location/Qualifiers
     source          1..4020
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1..17
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        22..40
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        98..116
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1598..1614)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1598..1614)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(1672..1690)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1695..1742
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1749..1766)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1749..1765)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1758..1780)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      1829..1915
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2007..2034
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2193..2210)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2364..2952)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2444..2463)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3099..3908)
                     /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     3822..3841
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3900..3919
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 caggaaacag ctatgaccat gtaatacgac tcactatagg ggatcctcaa aaaacccctc
       61 aagacccgtt tagaggcccc aaggggttat gtggaagtaa tacgactcac tatagggcga
      121 attgggtacc gggcccggcc aattctaccc gagcttcact tgatccttga ggtttttgac
      181 ccttcgccca aattttatag tatctgtttt ttcctgttgg ttctaatatc gttttcgctg
      241 gaatcacata ttttataacg attcttccat tatcagtatt tctatttaat acgtatccag
      301 atatatcttc ttgctgtgaa cctgtattct caaggattat aaattttcca tctggagaac
      361 attcttgtat tgtcacattt ccttttgcag tcctctgaaa cgaagttttt gcagacattt
      421 ctcctttcac aacatttgta atctgtgttt tcgtcattgg ggatgtatgt tgctgactaa
      481 tcgtagatct ggtcatatgg tcagtcaaat ccatactatg acctccttga ctgaattgag
      541 catctttacg gttcactctg tattcttctc cttcaagcat cttcctatag gctgctattt
      601 caagttccaa acttaatttg gcatcgagta aaagttgtaa ttccaaaaca atattttcta
      661 attctgcttg acgattcatc aattcctttc ttaaatgaat cttctctgat tcaaattctc
      721 tatgcttttc gtctatttct cgtttaagca tatctagcat tcgttcaaga tcgttgtttc
      781 ttaactctaa ttcaggtaac ctatttctca tttctaccaa ttgttccttt aatctcgcat
      841 tttcttccct taactgaact gtttcagcgc tttcttttgt ttgggtcgta cggaattctt
      901 gaattttaag agtgtattga gcttccattt ccatttgaag agtttccatt tttctatcat
      961 attcattttg aatatctcgt aaagcttgag ccatttcgtt tctccagaag tctttgctat
     1021 ccgaagcatc tttatatatc aatgatgata attctcgtat ttctgattca aagattttct
     1081 tcaaaaactc aatctgatct agaagagatt gaacttcatt ttgagcagta atatgttcaa
     1141 gtgtttcttt atcaagatcc ttcttcagtt gatttatttc ttctttcagt ctaaaaattt
     1201 ctttattgtc tcgaaccttt tcttcttcta atccagctaa tcttctgcga agactcgaaa
     1261 tttcgtgttc gtaatcggaa agttgatttg tcattttatc tattgtttcc ttatcggaaa
     1321 tgtggagttg attcacatca tttaattcta tttgagcttc ttgaatcaaa tcctctattg
     1381 aattaattcg catttctaat tgcgcttttg aagcttcagc ttgatccaaa gactttctga
     1441 gtaacaccat gtcattatca tacagtgctt tgattcttgc tgtctctttt ccccattggt
     1501 cttttaaatc cattaactcc tctgatagtc gtttattttc atcctccaga tgtcgcactt
     1561 tatcgggatg gtaatgccgc tagccatgga accggtggat ccactagttc tagagcggcc
     1621 gccaccgcgg tggagctcga attcatcgat gatatcagat ctgccggtct ccctatagtg
     1681 agtcgtatta cttcctagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     1741 tgcttaagac tggccgtcgt tttacaacgt cgtgactggg aaaacatcca tgctagcgtt
     1801 aacgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt cggaagactg
     1861 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc
     1921 gggagcggat ttgaacgttg tgaagcaacg gcccggaggg tggcgggcag gacgcccgcc
     1981 ataaactgcc aggcatcaaa ctaagcagaa ggccatcctg acggatggcc tttttgcgtt
     2041 tctacaaact cttcctggct agcggtacgc gtattaattg cgttgcgctc actgcccgct
     2101 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     2161 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     2221 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     2281 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     2341 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     2401 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2461 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2521 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2581 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2641 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2701 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2761 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     2821 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2881 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     2941 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgac
     3001 gcgtaactca cgttaaggga ttttggtcat gagcttgcgc cgtcccgtca agtcagcgta
     3061 atgctctgcc agtgttacaa ccaattaacc aattctgatt agaaaaactc atcgagcatc
     3121 aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt
     3181 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
     3241 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
     3301 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa
     3361 agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
     3421 tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg
     3481 cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact
     3541 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct
     3601 gtttttccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
     3661 ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta
     3721 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
     3781 ccatacaagc gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac
     3841 ccatataaat cagcatccat gttggaattt aatcgcggcc tcgacgtttc ccgttgaata
     3901 tggctcataa caccccttgt attactgttt atgtaagcag acagttttat tgttcatgat
     3961 gatatatttt tatcttgtgc aatgtaacat cagagatttt gagacacggg ccagagctgc
//
