LOCUS       sequence-209321-        4191 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 4191)
  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 4191)
  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_PC2
FEATURES             Location/Qualifiers
     source          1..4191
                     /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(1821..1839)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1897..1915)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1920..1936)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1920..1936)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2038..2057)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2049..2858
                     /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(2116..2135)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3005..3593
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3494..3513
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3747..3764
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3923..3950)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4042..4128)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4177..4191,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4191,1..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 cggtgagtgc cgtacctagt tggagccacc acatgaatct
      241 tgttgcgcat ccgatggtga aacaggccaa tcaagcggag ggttatcttc gtaagaagag
      301 gggctacaag ccagtcaaga gggatcgaat cccactacca agtataaaca agaaagggat
      361 cccagaccga gagaaggtca agaatgagaa tatcaagctc ccaaccgacc cattatttgc
      421 caaggaatgg tacattagga ataccgggca ggcagagggg gtcaaggacc ttgacctgaa
      481 cgttctctcg gcatgggcac aaggcatcac aggtaagggg gtgaccacag ctatcatgga
      541 cgacggtatc gactatctgc atccggattt gtctaagaac tacaatgccg agagtagcta
      601 tgacttcagt agtaatgatc cgtacccatt cccgcggtac actgatgact ggttcaatag
      661 tcacgggact agatgtgctg gggagatctc tgccgagagg gacaatggcg tctgcggagt
      721 ggggatcgct tatgactcca aggtggtcgg attacggatg ctcgatcaac catatatgac
      781 cgatctgatc gaatcagcgg cgatgagtca cgatcgcgac tcaatccata tatactcggc
      841 tagttggggc cccactgatg acgggagaac cgtggacggg cctcgaaacg agaccatgaa
      901 ggccatagtg gatggagtga acaagggtcg tcatgacaag ggtaatatat atgtgtgggc
      961 ctcaggggac ggaggagtgg ccgatgactg caactgtgac gggtacgccg ccagtatgtg
     1021 gaccattagc attaactccg ccaccaatga tggcgagact gcaagttatg acgagtcgtg
     1081 ctcctcgacc ctggcgtcca ccttctccaa tggcaaggca tggtacagtc tcgatgcagg
     1141 cgtaggtacc gtggatcttt atggacaatg caccctccac cattccggga catccgccgc
     1201 cgcacctgaa gcggccgcgg tgttcgcctt ggcgctcgaa gcgaatacga acattggatg
     1261 gcgagacatg cagcatctgt cagtgctgac aagcaagaga aaccatctgc atgaccggga
     1321 ccagcgccat aattggacga tcaatggcgc gggcctggaa ttcaatcact tattcgggta
     1381 cggagtgctg gacgcaggcg atatggtgcg ggtggctaag cagtgggtca atgtgccaga
     1441 gagattccat tgcattgcgg gtacaatcga caagaggatg atattcaatg tcagcaaggc
     1501 actggtgctc gagatagaca cggacgcctg taagttggac gaggcacaca tggtcaacta
     1561 cctggagcat gtgcaggcct tcgtcaccgt gaaggctagt cgccgtggcg acatcacaat
     1621 gttcctgacg agcccgatgg gcacgacctc gatgattctt agtcgccgcg agaaggacga
     1681 cgaccctagt cgcgggttca cgaagtggcc attcatgacc acgcacacct ggggtgagaa
     1741 ccctcgtggc aagtggcgac tgacagtaat gttcgatacc gaggacgggt agaattggcc
     1801 gggcccggta cccaattcgc cctatagtga gtcgtattac ttccacataa ccccttgggg
     1861 cctctaaacg ggtcttgagg ggttttttga ggatccccta tagtgagtcg tattacatgg
     1921 tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac
     1981 aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa
     2041 ggggtgttat gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg
     2101 atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa
     2161 tctatcgctt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta
     2221 gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc
     2281 ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg
     2341 cgatccccgg aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata
     2401 ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc
     2461 cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt
     2521 tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga
     2581 aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct
     2641 cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag
     2701 tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt
     2761 ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata
     2821 aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt
     2881 aacactggca gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc
     2941 ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3001 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3061 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3121 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     3181 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3241 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3301 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3361 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3421 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     3481 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     3541 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     3601 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     3661 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     3721 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     3781 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     3841 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt accgctagcc
     3901 aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg
     3961 atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac
     4021 gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
     4081 gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat gcctggcagt
     4141 tccctactct cgcgttaacg ctagcatgga tgttttccca gtcacgacgt t
//
