LOCUS       sequence-208834-        3411 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 3411)
  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 3411)
  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_1188_TNXB/p plate_10
FEATURES             Location/Qualifiers
     source          1..3411
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1..17
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(890..908)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(966..984)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(989..1005)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(989..1005)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1107..1126)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1118..1927
                     /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(1185..1204)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2074..2662
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2563..2582
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2816..2833
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2992..3019)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3111..3197)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3246..3268
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3260..3277
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3261..3277
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(3284..3331)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3336..3354
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 tctagaacta gtggatccac cggttccatg gctagcggca ttaccatccc gtgtctgtgc
       61 aaaagattcc tcaaaaagaa ataaaattgg cggtggagga ccgtgcagaa gaccggatga
      121 gtgctttttc gggacattgt gtaaagaaac gtcgcgctaa aattgggttt tgttccaaag
      181 atgcaccgac atttaaccag attggcggtg ggggtccttg catattccat acccaatgtt
      241 actccttcta ctgtcacaaa ctcaatgata agattaaagc atgtgccaag gattcggtat
      301 atcccgcgaa agtcggagga ggaggaccat gccgaaatga ttttgactgt gcttcgaatt
      361 tctgctacca agcaggggga gtcaaattct gtgttcagga gtcaaaatca agacttggat
      421 ttatctctga gggagggctg tgcaagtctc atagatcgtg taattctgga ttttgtgtga
      481 agaataattc aacgggtttc tgtgccaaag agtctcggat ggatgggaaa atcggaggag
      541 gtggtccttg caagatggac gatcaatgct tctctggcta ttgcttcaac aatggcagca
      601 tgaagtactg tgctaaggat ggtcataaat tcaacagagt gggtatcagt ggtccatgta
      661 gacgaaattc cgagtgcagc tctaaacggt gcgtagaaat gaacctaaag actgctatca
      721 acaacaacac caaagccaaa gtgtgtttgg aaaagaaaaa acatagcacg aaaaaaggga
      781 acacaattaa gtgaactaat ttcgtaattt gaaaaataat gatttaattt aataatgtgt
      841 aataattgtg gcggcgggta gaattggccg ggcccggtac ccaattcgcc ctatagtgag
      901 tcgtattact tccacataac cccttggggc ctctaaacgg gtcttgaggg gttttttgag
      961 gatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     1021 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     1081 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     1141 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     1201 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     1261 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     1321 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     1381 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     1441 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     1501 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     1561 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     1621 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     1681 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     1741 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     1801 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     1861 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     1921 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     1981 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     2041 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2101 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2161 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2221 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2281 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2341 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2401 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2461 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2521 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     2581 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     2641 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     2701 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     2761 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     2821 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     2881 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     2941 caacgcaatt aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca
     3001 tccgtcagga tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc
     3061 cgccaccctc cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta
     3121 ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga
     3181 gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat
     3241 gttttcccag tcacgacgtt gtaaaacgac ggccagtctt aagcaaaaaa cccctcaaga
     3301 cccgtttaga ggccccaagg ggttatgcta ggaagtaata cgactcacta tagggagacc
     3361 ggcagatctg atatcatcga tgaattcgag ctccaccgcg gtggcggccg c
//
