LOCUS       sequence-208121-        4243 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 4243)
  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 4243)
  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: Gsp_PC2
FEATURES             Location/Qualifiers
     source          1..4243
                     /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(1873..1891)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1949..1967)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1972..1988)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1972..1988)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2090..2109)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2101..2910
                     /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(2168..2187)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3057..3645
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3546..3565
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3799..3816
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3975..4002)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4094..4180)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4229..4243,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4243,1..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 cgatgtacgt cttcatgatc ccggaactca tgatctagcc
      241 aataaagtag ctcaagaaac tggtttctca ctattgagaa accatataca atctccaaat
      301 gaatttgttt accatcataa agctgtgccc catgtgagtt caattccaag ttggaatcat
      361 cataaagttt tattatctca tccacttgtt aaagaagcta aacaagttga aggttttatt
      421 agaaaaaaac gaggttttaa accgatcaaa aagcaagatc gtattccact tccaacaata
      481 aacaaaaaag gtattcctgg cagaggaaaa acagagaaca ttaaagaaaa tatgaaatta
      541 ccaactgatc ctttgttttc aaaagaatgg tacattcgaa acactggaca agctgatgga
      601 gtaaaagatc ttgatctaaa tgttttatcc gcatgggctc aaggcattac tggtaaaggt
      661 gttacaacag caattatgga tgacggtatc gattatcttc atccagattt atcgaaaaat
      721 tacaatgcag aaagtagtta tgatttcagt agtaacgatc catatccatt tccaaggtat
      781 actgatgatt ggtttaatag tcatggaaca agatgcgccg gtgaaatttc agctgagaga
      841 gacaatggaa tttgtggtgt tggtgtggcc tacaattcta agatcgccgg attgcgtatg
      901 ttggatcaac cgtatatgac agatcttatt gaagcagcag caatgagtta tgatagagat
      961 aatattgata tttattctgc cagttggggt cctacagatg atggtagaac agtcgatgga
     1021 ccaagaaatg agaccatgcg tgcaattgta gaaggcgtta ataaaggtcg taaaggtaaa
     1081 ggaagcattt acgtttgggc atctggtgat ggaggagcag ccgatgattg caattgcgac
     1141 ggatatgcag caagcatgtg gactattagt attaattctg caaccaatga tggagaaact
     1201 gcaagctacg atgaatcttg ttcttcaacc cttgcctcaa cattttcaaa tggaaaagca
     1261 tggtttagtc ttgacgcagg agttggtacg gttgatttat atggacaatg caccttacat
     1321 cattctggaa cttctgctgc ggccccggaa gcagctggag tatttgcttt agctttagaa
     1381 gctaattcta atattggttg gagagacatg caacatttaa ctgtgctcac tagtaaacga
     1441 aaccatttgc acgatcgaga tcgacgtcat aattggacaa ttaatggggc cggtttggaa
     1501 tttaatcatc tttttggtta tggagttctt gatgctggtg atatggttcg tgttgctaaa
     1561 aaatgggtaa atgttcctga aagatttcac tgtatagcgg gacaaattaa taaacgttat
     1621 cggtttaatg ttactaaagc actgatactc gagatagata ccgatgcttg caaatcagat
     1681 actggccatc gagttaatta tctagaacat gttcaagctt ttatatcagt aaaagctagt
     1741 agacgaggag atataacaat gtttcttacc agcccaatgg gcacaacatc tatgattctc
     1801 agtcatcgac caaaagacga tgattctaca cgaggatcgg gtagaattgg ccgggcccgg
     1861 tacccaattc gccctatagt gagtcgtatt acttccacat aaccccttgg ggcctctaaa
     1921 cgggtcttga ggggtttttt gaggatcccc tatagtgagt cgtattacat ggtcatagct
     1981 gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa
     2041 aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     2101 atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat
     2161 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc
     2221 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     2281 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     2341 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
     2401 ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     2461 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     2521 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     2581 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     2641 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     2701 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     2761 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     2821 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     2881 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg
     2941 cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg
     3001 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3061 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3121 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3181 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     3241 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3301 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3361 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3421 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3481 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3541 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3601 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3661 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     3721 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     3781 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     3841 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     3901 gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag
     3961 tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg
     4021 cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat
     4081 ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
     4141 gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact
     4201 ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac gtt
//
