LOCUS       sequence-209286-        3596 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 3596)
  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 3596)
  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_frz-4
FEATURES             Location/Qualifiers
     source          1..3596
                     /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(1226..1244)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1302..1320)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1325..1341)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1325..1341)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1443..1462)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1454..2263
                     /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(1521..1540)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2410..2998
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2899..2918
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3152..3169
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3328..3355)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3447..3533)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3582..3596,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3596,1..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 cggcagttgg actttttagt gggcaacggc aacgagacga
      241 cctggtgtaa gattaatttc ttgatgctct actttttcag ttcagccagc gcgttgtggt
      301 gggtcgtttt aacagtcacg tggtttcttt cggcatctcg gcattggggg tatgaggcta
      361 ttgaatcagt cagcagtatt ttgcatctag tcagttgggg aattccggcg ctcaagtcaa
      421 ttttcgttct catccttcac aaaatcgacg ccgatgaatt aactggacaa tgttacgtcg
      481 gcaactcaga taaggaagcg ctgattggtt tcattctagt cccacaagtt ctctacttag
      541 tgaccggggt cacgttccta gcgttgggct actactccat cttgaaggta agaaagtctt
      601 tcctacaaca ggtgagttgt attgacacca cgaatttgaa acgacttgaa aaattaatgg
      661 cgaaaatcgg cgtcttttcc attttatata ttgtaccagt cacgtgtaca attataagct
      721 acgttatcga catcttcagg acttcctatt tcgatttaat gctgaaggcg ttgtatgaat
      781 attcagcaga ttgtgtcaca gtaaagggtc tgtcgtggtc gttggcgaag tgcgtagaca
      841 tcattcaacc ggcattgccg tccatcgaag tgaggatgct gcgtattttt atgaatcttg
      901 tcatcggcat aacatcgggc atttggattt ggggtaacaa aaagtcgttc cgatattgtg
      961 ggcgaaataa aaaagaaatt caacaatgtg aaatgagaca aatcgaagta ttaacccccc
     1021 tcaatcagac tccgtttatt aaatccggtg aggacacttt tccccatcac gggtggagtt
     1081 atgtgccaac cttggcctcg gaacacgcgg tcggggggct ctctcagacc cctgtggaag
     1141 gtataatggc ccccaataac tcatcaataa gcgaagtatc cagtaagcat cgggtagaat
     1201 tggccgggcc cggtacccaa ttcgccctat agtgagtcgt attacttcca cataacccct
     1261 tggggcctct aaacgggtct tgaggggttt tttgaggatc ccctatagtg agtcgtatta
     1321 catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat
     1381 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa
     1441 tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa
     1501 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     1561 gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     1621 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     1681 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     1741 cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     1801 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     1861 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     1921 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     1981 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     2041 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     2101 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     2161 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     2221 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg
     2281 gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa
     2341 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     2401 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     2461 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     2521 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     2581 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     2641 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     2701 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     2761 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2821 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2881 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2941 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     3001 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     3061 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     3121 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     3181 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     3241 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
     3301 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
     3361 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
     3421 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
     3481 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
     3541 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgtt
//
