LOCUS       sequence-208379-        3436 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 3436)
  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 3436)
  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_sfrp-1
FEATURES             Location/Qualifiers
     source          1..3436
                     /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(1066..1084)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1142..1160)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1165..1181)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1165..1181)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1283..1302)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1294..2103
                     /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(1361..1380)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2250..2838
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2739..2758
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2992..3009
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3168..3195)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3287..3373)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3422..3436,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3436,1..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 cgtcgttcca aagtggttac acaattgatc aatgctatca
      241 tattcctgat aatttcagtc tttgtcgtaa tgttggttat aatagaatgg ttttaccaaa
      301 ttacattcaa catgaatcta ttgaagaagc tgtacaacat tctcgagttt ggatgggttt
      361 agtaaataca cagtgtcata ttgatatcag aaaattcatt tgttctcttt atgctccggt
      421 atgtattagc agtcaaagag ttcaaaaggt tccaccttgt agggaattat gtgtaactgt
      481 aaaggaatct tgtttaccaa gtatgaaaat gtatggattt gattggccaa caataatgaa
      541 atgttcaaag tttccacaat ttcaaaattc tctatgtata ccaagaactc tcgttccaga
      601 aaagaactgt gatttatgca taaaaaatcc atcatatgaa gttatagcaa atagattttg
      661 tttatcaccg atagttattc gagcaaaaat caaacgaatt attccgattg gtagtaaaac
      721 aatgcagata gtattacaga aaaaatcgaa atttctcaaa ttttttgcta atttcagtca
      781 aaccgatttg aatttacgtt tgaaatgtaa ctgcacaaat ttaaaattca aacaacgagc
      841 attgaaaggt cgatggataa taatggctaa agtaaatcaa aatgaagctg ttgtaaattt
      901 cattactaaa tggaaaagga aaaaagctga attcaaacat tcaatgaaaa tcatacagaa
      961 ttttggacat attgtgtgca aatcaaaatt accagatggg attagtttac gagaacgata
     1021 tgctgcctat cgggtagaat tggccgggcc cggtacccaa ttcgccctat agtgagtcgt
     1081 attacttcca cataacccct tggggcctct aaacgggtct tgaggggttt tttgaggatc
     1141 ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct
     1201 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg
     1261 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg
     1321 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     1381 aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag
     1441 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     1501 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     1561 gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa
     1621 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     1681 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
     1741 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     1801 ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat
     1861 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     1921 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     1981 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     2041 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     2101 taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac
     2161 ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca
     2221 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2281 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2341 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     2401 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2461 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2521 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2581 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2641 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2701 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     2761 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     2821 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     2881 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     2941 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3001 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     3061 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     3121 gcaattaata cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa aggccatccg
     3181 tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc
     3241 accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca
     3301 ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct
     3361 ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt
     3421 tcccagtcac gacgtt
//
