LOCUS       sequence-209328-        4054 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 4054)
  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 4054)
  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: Pgr_ifb
FEATURES             Location/Qualifiers
     source          1..4054
                     /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(1632..1648)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1632..1648)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(1706..1724)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1729..1776
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1783..1800)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1783..1799)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1792..1814)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      1863..1949
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2041..2068
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2227..2244)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2398..2986)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2478..2497)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3133..3942)
                     /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     3856..3875
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3934..3953
                     /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 ggacgattct tccgttatca gtgtttctgt
      181 taactttgaa tccggaaata ttttctggct gccctcctgt attctcaagt acgacaaact
      241 tcccatctgg ggaacactcc tgaattgtca catttccctt tgcagttctt tgaaatgaag
      301 tcttcgcaga catttgacct ttcacagtgc tgctcacttg ggttttgctt gttgtgaccg
      361 cagccatcga ttgacccacg tttgtacgag ttatcgtcga gtctgcaaaa tccattcgtt
      421 gaccactact ggatgacatc gcgttggatg acatagcatt taatgacatc gcattctttt
      481 gattgactct gtgctcctcg ccttcgagca tcttccggta ggccacaatt tctaattcta
      541 aacttaattt agcgtccaac aaaatttgta gttcagcgaa aatgctctca agttgtgctt
      601 gtcgacttga taattccttt cttagatttg ccctttccat atcgtagtca cgacttcttt
      661 cgtcgatctc tcgtcgtaac atttccagca ttctttccag ttcgttgttc ctggcttcta
      721 gttccggtag tcggttacgc atttccgaca gttgttcctt caatttggtg ttgtcttcgc
      781 gcaactgaat tgtctcctgg gtgtcctttg actgggttat tcgatagtcc tggatcttaa
      841 gattgtattg cgcctccata tcgtgttgta ttgagtcaat gcggcgatcg tattctgatt
      901 gaatgtcccg caacgcctga gccatttcat tcctccagaa gtccttactc tcggaggggt
      961 ccttgtaggc caaagatgtc aattctctga cctcggattc atatatcttc ttcaggaact
     1021 ctatcttgtc tcgtaacgat tgaacctcat tttgagccat aatatgatca agggtttctt
     1081 tgtcgagatc ctttttcaac tgagcaactt cgtctttcag acgggaaatt tctttgtcgt
     1141 ctcttatttt ttcgtcttcc aagccggtta aacgtcttcg aagactcgta atctcgtttt
     1201 catagtccga cagttgcgaa ttcatcttgt caatggtttc tttatccgca ttatgaactt
     1261 gattcacttc gctcaactca atttccgact cctgaagaaa atcttcaaac gaattgattc
     1321 tcatctccaa ctggaccttt gcaccttctg cggagtctaa ggattttctt agattgttca
     1381 tatcattatc atattgatct ttgattctct ccgtctcttt gccccattga ttttttagtt
     1441 ctgttaattc gtcagtgagt cgtctatttt cgtcctcgag atgcttcact ttgtcaatgt
     1501 aattggcaaa ccgatcattc aactcctgga gttcatgtcg ttctttctct cgtccgttca
     1561 tcagactact cacgccactc tggacaatat tggacgtcgg gatggtaatg ccgctagcca
     1621 tggaaccggt ggatccacta gttctagagc ggccgccacc gcggtggagc tcgaattcat
     1681 cgatgatatc agatctgccg gtctccctat agtgagtcgt attacttcct agcataaccc
     1741 cttggggcct ctaaacgggt cttgaggggt tttttgctta agactggccg tcgttttaca
     1801 acgtcgtgac tgggaaaaca tccatgctag cgttaacgcg agagtaggga actgccaggc
     1861 atcaaataaa acgaaaggct cagtcggaag actgggcctt tcgttttatc tgttgtttgt
     1921 cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgtgaagc
     1981 aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaactaagc
     2041 agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttcct ggctagcggt
     2101 acgcgtatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     2161 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     2221 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2281 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2341 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2401 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2461 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2521 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2581 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2641 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2701 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2761 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2821 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2881 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2941 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3001 cttttctacg gggtctgacg ctcagtggaa cgacgcgtaa ctcacgttaa gggattttgg
     3061 tcatgagctt gcgccgtccc gtcaagtcag cgtaatgctc tgccagtgtt acaaccaatt
     3121 aaccaattct gattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc
     3181 aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc
     3241 gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac
     3301 atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc
     3361 atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg
     3421 ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt
     3481 cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca
     3541 aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc
     3601 tgaatcagga tattcttcta atacctggaa tgctgttttt ccggggatcg cagtggtgag
     3661 taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc
     3721 cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc
     3781 atgtttcaga aacaactctg gcgcatcggg cttcccatac aagcgataga ttgtcgcacc
     3841 tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga
     3901 atttaatcgc ggcctcgacg tttcccgttg aatatggctc ataacacccc ttgtattact
     3961 gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta
     4021 acatcagaga ttttgagaca cgggccagag ctgc
//
