LOCUS       sequence-208377-        4057 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 4057)
  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 4057)
  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_ifb
FEATURES             Location/Qualifiers
     source          1..4057
                     /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(1687..1705)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1763..1781)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1786..1802)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1786..1802)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1904..1923)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1915..2724
                     /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(1982..2001)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2871..3459
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3360..3379
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3613..3630
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3789..3816)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3908..3994)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4043..4057,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4057,1..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 cgtcagtggg gtaaagagac tgccagaatc aaagctttat
      241 atgattctga tatgagtcaa ttaaggcgtt cattggatca agctgaagca tcaaaagctc
      301 aacttgaaat gaaaataaac actttagagg atcttattca agaagctcaa atggaattaa
      361 atgatgctac ccagttgcat tctgctgata aagaaacaat agataaaatg agcagtactt
      421 tggctgacta tgaacatgaa attggaaatc ttcgaaaaag attgagctca ttagaggaag
      481 aaaaaattag agataataaa gaaataattc gattaaaaga agagcttaat caattgaaaa
      541 aggatttgga taaagaaact ctagaacata ttactgctca taatgaagtt caatcattac
      601 tcgatcaaat cgaatttttg aagaaaatat tcgaatctga aataagagaa ttggctgcat
      661 tggtttataa ggattcctct gaaagtaaag atttctggag aaatgaaatg gcccaagctt
      721 taagagatat tcagtcggaa tacgataaaa aaatggaagc tttgcactat gatatggaag
      781 cacaatatac tcttaaaata caagaaattc gaacaaatca aagcaaagaa aacgttgaga
      841 cagtacaatt aagagaggaa aatactagat tgaaagatca gttagcagat atgagaaata
      901 gacttccaga attagaagct agaaatagtg aattggaaag attattagaa ttgttgagac
      961 gagaaattga agaaaagaat cgtgaatttg atatggagaa agctaattta aggaaagaac
     1021 tgatgaatcg acaatcggaa cttgaatcaa tacttataga attgcaaatt ttactagatg
     1081 ccaaattaag cttagaactt gaaattatag cttatagaaa aatgttggaa ggtgaagaat
     1141 atagagtcaa tcgtaaaaat aatatcagta gtagtaattc atttaaccaa gcaggaagaa
     1201 atgttgattt gatggatcat gtaacaagat ccagtatcag tcaacaacaa gctgctcctt
     1261 caactaaaac tcaagttacg agtattgtca aaggagaaat gtcagcaaag acgtcttttc
     1321 aaagaactgc aaaaggtaat gtaacaattc aagaatgttc ggctgatgga aaatttatta
     1381 ttcttgaaaa tactggatca caagatgagg atatttccgg ttatattctc aacagaaata
     1441 ctgacaatgg acgaattatt gtcaagcatg tgatttcacc aaaaactatt ttagaaccca
     1501 caggaaaaaa tcgatattat aaaatatggg caagaggaaa taaacctcaa ggttcaaatg
     1561 atgcagaatg ctcagaatcg aattggggat acggagctca gattgttacg acattgcaaa
     1621 attcaaaagg cgaagaaaag gcaactcaca ccgggtagaa ttggccgggc ccggtaccca
     1681 attcgcccta tagtgagtcg tattacttcc acataacccc ttggggcctc taaacgggtc
     1741 ttgaggggtt ttttgaggat cccctatagt gagtcgtatt acatggtcat agctgtttcc
     1801 tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata
     1861 tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
     1921 catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat
     1981 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat
     2041 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     2101 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     2161 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa
     2221 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     2281 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     2341 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     2401 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
     2461 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     2521 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     2581 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     2641 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     2701 ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc
     2761 attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac
     2821 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2881 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2941 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3001 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     3061 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3121 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3181 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3241 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3301 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3361 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3421 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3481 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3541 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3601 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     3661 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     3721 aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta
     3781 gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt
     3841 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc
     3901 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
     3961 cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg
     4021 ttaacgctag catggatgtt ttcccagtca cgacgtt
//
