LOCUS       sequence-208836-        3908 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 3908)
  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 3908)
  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_collagen
FEATURES             Location/Qualifiers
     source          1..3908
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(168..756)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(248..267)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(903..1712)
                     /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     1626..1645
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1704..1723
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1825..1841
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1825..1841
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1846..1864
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1922..1940
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3310..3326)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3310..3326)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(3384..3402)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      3407..3454
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3461..3478)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3461..3477)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3470..3492)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      3541..3627
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3719..3746
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(join(3905..3908,1..14))
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
       61 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
      121 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
      181 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
      241 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
      301 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
      361 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
      421 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
      481 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
      541 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
      601 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
      661 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
      721 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
      781 gggtctgacg ctcagtggaa cgacgcgtaa ctcacgttaa gggattttgg tcatgagctt
      841 gcgccgtccc gtcaagtcag cgtaatgctc tgccagtgtt acaaccaatt aaccaattct
      901 gattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
      961 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     1021 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     1081 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     1141 actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc
     1201 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     1261 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     1321 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     1381 tattcttcta atacctggaa tgctgttttt ccggggatcg cagtggtgag taaccatgca
     1441 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     1501 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     1561 aacaactctg gcgcatcggg cttcccatac aagcgataga ttgtcgcacc tgattgcccg
     1621 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     1681 ggcctcgacg tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa
     1741 gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta acatcagaga
     1801 ttttgagaca cgggccagag ctgccaggaa acagctatga ccatgtaata cgactcacta
     1861 taggggatcc tcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgtggaa
     1921 gtaatacgac tcactatagg gcgaattggg taccgggccc ggccaattct acccggttgg
     1981 tgatctcatt tgcatggcca ttactccacc tggtggtcca gggggccctg gtggaccaat
     2041 aggacccctc tggccaatag ggccactttc tccttgtctt cccggtaatc cggcaacacc
     2101 tataggtcca agaacaccag gatctccgcg tggaccaatc actcccgtgg gaccagttgg
     2161 tccctctgta cctgatggac caggaggtcc gtcgggacct gagaaaccag ttggtcctct
     2221 gtcaccatct gatcctgttg gtccatcaaa gccgggactt cctttaagtc ctttttttcc
     2281 cggaagccct cgtgaacctg gacgtccagc ttgtcctggt tttccatttg ctcctgaaac
     2341 tcctttatcg ccgaccaagc cagcttctcc taatgctcca ttaggtccta gttccccagc
     2401 atctcctgga tcgccagagg gaccgggttt acccagagca cctctagaac caggcactcc
     2461 tggaaatcct ccagatccag cttttccagg tttccctttt gcaccaggtt tgccttttcc
     2521 acccggtccc ccaggtgggc ctggtgctcc aacagttcca ggatttccgg gtggaccatc
     2581 cattccaact ggcccgggaa caccaagttc gccataatcg ccagttaaac cctcactacc
     2641 agggagacct ggttttcctg gtttacctag acttccttgt gggccagaag gaccaggcga
     2701 tcccattact ccggggtctc ctgtggttcc atcagggcct ggcttgccat cagttccagg
     2761 agctcctgcc tttccctttt cgccctgtgg acctctaaga cccaatggac ctttgcgtcc
     2821 atttgctcca ttcattccag gtactcctgg aacgcctagt cctccaggtt ttccaactgg
     2881 tccttcttta ccaggatcgc cttgctttcc ttgttttcca ggtaatccta gatttccatc
     2941 tcgtcctggt aaaccaactg gtccaggagc tccggaaata ccaggatctc ctgcttctcc
     3001 ggcaggacca ggggcaccta cacttccttc taatcccatt ggtcctaaaa gtcctggctt
     3061 tccaggagct cccagggctc ctgatttccc ttcttcaccg tcaggcccag gagctccggc
     3121 atcgccagga tcaccagcat ctccatcagg accagtagcg ccaggtttac cacttggacc
     3181 aacttctcct ggaacgccat caagccccgg tttaccagtt tctcccttgg gacctgcagg
     3241 gcctgctccc ctgggctgtg gtttgcttat tcgagcggga tggtaatgcc gctagccatg
     3301 gaaccggtgg atccactagt tctagagcgg ccgccaccgc ggtggagctc gaattcatcg
     3361 atgatatcag atctgccggt ctccctatag tgagtcgtat tacttcctag cataacccct
     3421 tggggcctct aaacgggtct tgaggggttt tttgcttaag actggccgtc gttttacaac
     3481 gtcgtgactg ggaaaacatc catgctagcg ttaacgcgag agtagggaac tgccaggcat
     3541 caaataaaac gaaaggctca gtcggaagac tgggcctttc gttttatctg ttgtttgtcg
     3601 gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgtgaagcaa
     3661 cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca aactaagcag
     3721 aaggccatcc tgacggatgg cctttttgcg tttctacaaa ctcttcctgg ctagcggtac
     3781 gcgtattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3841 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3901 cttcctcg
//
