LOCUS       sequence-208385-        3609 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 3609)
  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 3609)
  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: Ddo_collagen
FEATURES             Location/Qualifiers
     source          1..3609
                     /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(1239..1257)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1315..1333)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1338..1354)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1338..1354)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1456..1475)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1467..2276
                     /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(1534..1553)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2423..3011
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2912..2931
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3165..3182
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3341..3368)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3460..3546)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3595..3609,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3609,1..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 cggacctcaa ggaatagctg gtaaacctgg taaaccaggt
      241 ccaactggag gtataggttt acagggagac catggtgaaa tgggaaaacc tggtcctcaa
      301 ggaccacttg gtgtacctgg tcttactgga ccagtaggac caagaggtga tccgggacca
      361 cctggaagta aaggtgcagc aggtaaacct gggaaagatg gaactagtgg tggtaaaggg
      421 tacgatggac aaaccggaac tagtggcaaa tcgggtaaaa ttggagctcc cggtgaaaag
      481 ggagcagatg gaaaaccagg gccaaatggt cttcttggcg atactggttt aagaggtgca
      541 aaaggaaaaa aaggaacaga tggaactcca ggtagggaag gaaaaccggg tcctcgagga
      601 gctaaaggaa ctaaaggtcc aaaaggagtt cctggatatc ccggaagacc aggaattgaa
      661 ggagatccag gacctccagg ttattccgga cccgatggac caacaggccc taaaggccct
      721 gacggtttac aaggaccagt tggaatattc ggacctcgtg gagaccctgg aataccaggt
      781 gcaattggac ctactggacc accaggtaaa ccaggtggaa ctggtcctgc tggccaaaga
      841 ggaccgttgg gaccaccagg tccacctgga cctccaggag gcgtaatggc aatgcagatg
      901 cgatcaccca ctaaaggtgt tgtatacgga gatgatccga aggcagcaga actccttgga
      961 aataatgcta ttaaaaatcc tcaaggaaca aaagaagtac cagctagaac ctgcaaacat
     1021 cttagttcag taaatcctca agcacctgat ggtgagtact ggatagatcc gaatggaggt
     1081 cgaatagaag atgccgttaa ggtttattgt aaaatttctg agcaaaaaac ctgtatcaaa
     1141 tccattgata ataatctcag cattaggtca tggaatagtc caatacttaa tggtccgaca
     1201 tggcgggtag aattggccgg gcccggtacc caattcgccc tatagtgagt cgtattactt
     1261 ccacataacc ccttggggcc tctaaacggg tcttgagggg ttttttgagg atcccctata
     1321 gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc
     1381 tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt
     1441 acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc
     1501 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
     1561 ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc
     1621 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact
     1681 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg
     1741 catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc
     1801 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga
     1861 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat
     1921 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc
     1981 ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg
     2041 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
     2101 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga
     2161 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg
     2221 ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag
     2281 aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa
     2341 gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg
     2401 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2461 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2521 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     2581 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2641 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     2701 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     2761 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     2821 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     2881 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     2941 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3001 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3061 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     3121 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     3181 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     3241 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     3301 atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat
     3361 ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc
     3421 gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc
     3481 gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt
     3541 tatttgatgc ctggcagttc cctactctcg cgttaacgct agcatggatg ttttcccagt
     3601 cacgacgtt
//
