LOCUS       sequence-208830-        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     complement(25..42)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(196..784)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(276..295)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(931..1740)
                     /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     1654..1673
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1732..1751
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1853..1869
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1853..1869
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1874..1892
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1950..1968
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3450..3466)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3450..3466)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(3524..3542)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      3547..3594
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3601..3618)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3601..3617)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3610..3632)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      3681..3767
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3859..3886
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
       61 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
      121 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      181 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      241 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      301 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      361 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      421 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      481 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      541 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      601 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
      661 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      721 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      781 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg acgcgtaact
      841 cacgttaagg gattttggtc atgagcttgc gccgtcccgt caagtcagcg taatgctctg
      901 ccagtgttac aaccaattaa ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa
      961 ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa
     1021 tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc
     1081 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt
     1141 atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg
     1201 catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc
     1261 atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct
     1321 gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc
     1381 atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg ctgtttttcc
     1441 ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt
     1501 cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt
     1561 ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa
     1621 gcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa
     1681 atcagcatcc atgttggaat ttaatcgcgg cctcgacgtt tcccgttgaa tatggctcat
     1741 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt
     1801 tttatcttgt gcaatgtaac atcagagatt ttgagacacg ggccagagct gccaggaaac
     1861 agctatgacc atgtaatacg actcactata ggggatcctc aaaaaacccc tcaagacccg
     1921 tttagaggcc ccaaggggtt atgtggaagt aatacgactc actatagggc gaattgggta
     1981 ccgggcccgg ccaattctac ccgagcttca cttgatcctt gaggtttttg acccttcgcc
     2041 caaattttat agtatctgtt ttttcctgtt ggttctaata tcgttttcgc tggaatcaca
     2101 tattttataa cgattcttcc attatcagta tttctattta atacgtatcc agatacatct
     2161 tcttgctgtg aacctgtatt ctcaaggatt ataaattttc catctggaga acattcttgt
     2221 attgtcacat ttccttttgc agtcctctga aacgaagttt ttgcagacat ttctcctttc
     2281 acaacatttg taatctgtgt tttcgtcatt ggggatgtat gttgctgact aatcgtagat
     2341 ctggtcatat ggtcagtcaa atccatacta tgacctcctt gactgaattg agcatcttta
     2401 cggttcactc tgtattcttc tccttcaagc atcttcctat aggctgctat ttcaagttcc
     2461 aaacttaatt tggcatcgag taaaagttgt aattccaaaa caatattttc taattctgct
     2521 tgacgattca tcaattcctt tcttaaatga atcttctctg attcaaactc tctatgcttt
     2581 tcgtctattt ctcgtttaag catatctagc attcgttcaa gatcgttgtt tcttaactct
     2641 aattcaggta acctatttct catttctacc aattgttcct ttaatctcgc attttcttcc
     2701 cttaactgaa ctgtttcagc gctttctttt gtttgggtcg tacggaattc ttgaatttta
     2761 agagtgtatt gagcttccat ttccatttga agagtttcca tttttctatc atattcattt
     2821 tgaatatctc gtaaagcttg agccatttcg tttctccaga agtctttgct atccgaagca
     2881 tctttatata tcaatgatga taattctcgt atttctgatt caaagatttt cttcaaaaac
     2941 tcaatctgat ctagaagaga ttgaacttca ttttgagcag taatatgttc aagtgtttct
     3001 ttatcaagat ccttcttcag ttggtttatt tcttctttca gtctaaaaat ttctttattg
     3061 tctcgaactt tttcttcttc taatccagcc aatcttctgc gaagactcga aatttcgtgt
     3121 tcgtaatcgg aaagttgatt tgtcatttta tctattgttt ccttatcgga aatgtggagt
     3181 tgattcacat catttaattc tatttgagct tcttgaatca aatcctctat tgaattaatt
     3241 cgcatttcta attgcgcttt tgaagcttca gcttgatcca aagactttct gagtaacacc
     3301 atgtcattat catacagtgc tttgattctt gctgtctctt ttccccattg gtcttttaaa
     3361 tccattaact cctctgatag tcgtttattt tcatcctcca gatgtcgcac tttatcggga
     3421 tggtaatgcc gctagccatg gaaccggtgg atccactagt tctagagcgg ccgccaccgc
     3481 ggtggagctc gaattcatcg atgatatcag atctgccggt ctccctatag tgagtcgtat
     3541 tacttcctag cataacccct tggggcctct aaacgggtct tgaggggttt tttgcttaag
     3601 actggccgtc gttttacaac gtcgtgactg ggaaaacatc catgctagcg ttaacgcgag
     3661 agtagggaac tgccaggcat caaataaaac gaaaggctca gtcggaagac tgggcctttc
     3721 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg
     3781 atttgaacgt tgtgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg
     3841 ccaggcatca aactaagcag aaggccatcc tgacggatgg cctttttgcg tttctacaaa
     3901 ctcttcctgg ctagcggtac gcgtattaat tgcgttgcgc tcactgcccg ctttccagtc
     3961 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
//
