LOCUS       sequence-209334-        4186 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 4186)
  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 4186)
  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_frz-4
FEATURES             Location/Qualifiers
     source          1..4186
                     /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(1764..1780)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1764..1780)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(1838..1856)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1861..1908
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1915..1932)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1915..1931)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1924..1946)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      1995..2081
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2173..2200
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2359..2376)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2530..3118)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2610..2629)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3265..4074)
                     /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     3988..4007
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4066..4085
                     /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 gagatccata ttccagacgt gatcccaatg
      181 acaagattca tgaatattct tagcatcctc atctccacag atggcagcgc gggctggatt
      241 atatcgacac acttggccaa ggaccaggac agacctttga aagtcacaca atctgaggag
      301 tattcataga gggccttcag cattacgtca aagtatgggg tcttgaagac gtctataaca
      361 tagctgacaa tagtgcaagt cactggcact atgtaaagaa tggagaaaac cccaattttt
      421 gccatcaact tctcaagccg ctttaaattt gacggatcta tacaactagg ctgtctcaga
      481 aatgattttc tgaccttgag tattgagtag tatcctagtc ctagaaatgt gaccccaatg
      541 agcaaatata cgatttgcgg gaccaatata aagccaatca aggcgtcttt gtcgttgttg
      601 ccaacaaaac actggccagt caattcatcg gcgtcgattt tatggagaat aagcacaaaa
      661 atggatttca gtgctggaat accccaacta atgagatgta gtacactgct gatggactca
      721 atggcctcgt agccccaatg tctagacgct gaaagaaacc acgtgaccgt taacatcacc
      781 caccacaggg cgctggcaga actgaagaag tacaacatta ggaagttaat cttacaccag
      841 gtcgtctcat tgccataacc gaccaagtag tccaactgct cgtcgaacgg acgacaaatg
      901 acgttccgtc gccccaatat agcacctaaa agactgccgc aagaatggaa gaagtagcac
      961 attgacagga agacaattgg cttctctgga tacatgaatc ggttagtgtt gcaggaaaac
     1021 gtgatgatgg taaagagtga cgatatcaaa cacaagaggc tccatccaat cagccaaata
     1081 ctggcaaatg tcttatcgtc taggttgaac cgtatatcgg cgtcacaccg atgcatacat
     1141 gttgtgttcg gcaactgacc catattgatc tgagatggta gacacattgt acttgcaaat
     1201 gaaccggcct tcttcaacaa ttccatttgg tgcatgtttt tgtcattttt ctttcgtgcc
     1261 ttatccagtt gctggaagag ttctttcatt gtctctgaat tatttggctc tttggtgact
     1321 tctttgtctt gactgaatga atctggggtc aagcacattt tcccggttcc aggaagtttc
     1381 tcacagtcca gttcattagg ccacggtaca ttgtaagtgc tcgtcataaa gggctcacag
     1441 cgggacttca catgctgaca gaaccgttga catggttgca ccgatggggt cgatgtcgca
     1501 gcttccgcat cacaaatggg aaagtacatc gaacagagaa ataatttcaa ataatgactg
     1561 cattggagat taatcaacgg atagtagtcc cggactcgtt tgctagcctc attctgggaa
     1621 taaattccca tttgattggg cattctcgta tgagtatacc tgacgtcctg gcaaaccggt
     1681 tccgtgatca ctgtacactg cttgtacacg tcattcgatc ctttctcagc gggatggtaa
     1741 tgccgctagc catggaaccg gtggatccac tagttctaga gcggccgcca ccgcggtgga
     1801 gctcgaattc atcgatgata tcagatctgc cggtctccct atagtgagtc gtattacttc
     1861 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct taagactggc
     1921 cgtcgtttta caacgtcgtg actgggaaaa catccatgct agcgttaacg cgagagtagg
     1981 gaactgccag gcatcaaata aaacgaaagg ctcagtcgga agactgggcc tttcgtttta
     2041 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
     2101 acgttgtgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc
     2161 atcaaactaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttc
     2221 ctggctagcg gtacgcgtat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     2281 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2341 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     2401 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2461 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2521 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2581 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2641 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2701 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2761 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2821 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2881 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2941 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3001 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3061 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3121 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgacgcgt aactcacgtt
     3181 aagggatttt ggtcatgagc ttgcgccgtc ccgtcaagtc agcgtaatgc tctgccagtg
     3241 ttacaaccaa ttaaccaatt ctgattagaa aaactcatcg agcatcaaat gaaactgcaa
     3301 tttattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
     3361 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc
     3421 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag
     3481 tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc
     3541 tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac
     3601 caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa
     3661 aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac
     3721 aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt ttccggggat
     3781 cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag
     3841 aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac
     3901 gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaagcgata
     3961 gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc
     4021 atccatgttg gaatttaatc gcggcctcga cgtttcccgt tgaatatggc tcataacacc
     4081 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc
     4141 ttgtgcaatg taacatcaga gattttgaga cacgggccag agctgc
//
