LOCUS       sequence-208384-        3492 bp    DNA     circular SYN 15-APR-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 3492)
  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 3492)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_innexin-10
FEATURES             Location/Qualifiers
     source          1..3492
                     /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(1122..1140)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1198..1216)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1221..1237)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1221..1237)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1339..1358)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1350..2159
                     /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(1417..1436)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2306..2894
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2795..2814
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3048..3065
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3224..3251)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3343..3429)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3478..3492,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3492,1..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 cgggaacaat ctcgatcata gctatggttc tgtcagagtt
      241 cattgattac gctaccaaat acaaatgggt ggattttgtg ggcatggaag actttggcga
      301 ccggttaagt ttcaaccttt ccacttcgat attattgctt ctcactgttg ttatatcagt
      361 taaaagttat ttgttagctt caattgcttg ttatattcct attcaaccaa ccggtcatga
      421 tttcgatgat tttttgaaga gttattgttg ggtacatgga accataccga tgcataataa
      481 tcgtcccatg ccacgaactg aaaatgaatg ggaatatgca gacaaaatgg aaagaataaa
      541 ttattatcaa tgggttcctt ttgtacttgc tttacaaggg gtcctttttt atttaccgag
      601 aatcatatgg caaatgatca cttacaactt atctgggaca gatctggaag gactcgtgga
      661 aatggcgaat gctgctagta atcaaactgg tcaagcgaaa gcagaaactg taaatcaagt
      721 cgcaatgaat ctggaaaatc tgttgttttc ccatagaaaa attcgaaatc atagaaaagt
      781 aataatgttg aaactttctg aagctaaatc ttggttttgt gctggaaaaa gaagtggtcg
      841 gcaattaatc gttcaatata tgttcattaa attcttatat ttaattaatg ctttcggaca
      901 acttatactc atgcaatact tcttgaattt cggtcacaat gactcgagct ttggttacac
      961 ggttcttaag aacatactaa acgggaaaga atggaatgat actttaatat ttccaagggt
     1021 cacatattgt tatgtaaaaa atattaaaca tgttggaata acaaatgatt acttagctca
     1081 atgcgccggg tagaattggc cgggcccggt acccaattcg ccctatagtg agtcgtatta
     1141 cttccacata accccttggg gcctctaaac gggtcttgag gggttttttg aggatcccct
     1201 atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa
     1261 atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg
     1321 cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc
     1381 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg
     1441 tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt
     1501 ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa
     1561 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg
     1621 atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat
     1681 atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt
     1741 cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc
     1801 aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct
     1861 ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag
     1921 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag
     1981 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat
     2041 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta
     2101 ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat
     2161 cagaattggt taattggttg taacactggc agagcattac gctgacttga cgggacggcg
     2221 caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc
     2281 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2341 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2401 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     2461 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2521 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     2581 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     2641 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     2701 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     2761 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     2821 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     2881 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     2941 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     3001 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     3061 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     3121 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     3181 ttaatacgcg taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag
     3241 gatggccttc tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc
     3301 tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag
     3361 agcgttcacc gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg
     3421 ttttatttga tgcctggcag ttccctactc tcgcgttaac gctagcatgg atgttttccc
     3481 agtcacgacg tt
//
