LOCUS       sequence-208382-        3747 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 3747)
  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 3747)
  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_opsin
FEATURES             Location/Qualifiers
     source          1..3747
                     /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(1377..1395)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1453..1471)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1476..1492)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1476..1492)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1594..1613)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1605..2414
                     /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(1672..1691)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2561..3149
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3050..3069
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3303..3320
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3479..3506)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3598..3684)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3733..3747,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3747,1..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 cgcgttcatc cttactggag agaattcgaa ttggttccag
      241 aagtgtatca ttacttagtc ggagtttata tttcctttgt cggaatagca ggagttttag
      301 gaaatcttct agtactttat atatttgcta gtgcgaaaag tttgaggacc cctcctaata
      361 tgtttattat gagtttggcg atcggagatc tgacattttc agcagtgaat ggctttccac
      421 tattaactat ttcgagtttt aacactcgtt gggcttgggg aaaactaact tgtgaaatat
      481 acggattcat tgggggtctt ttcggattta tatccattaa tacaatggca cttatttcgt
      541 tggacagata ctttgttatc gctcaacctt ttcaaactat gaaatcgttg accattaaaa
      601 gagcgataat aatgctcgtg ttcgtttggc tttattcttt gatatggtcg acaccgccat
      661 tttttggatt tggaaactat gtccctgaag gatttcaaac ttcctgtaca tttgattatc
      721 taactcaatc caaaggaaac ataattttta atataggaat gtacatcggg aatttcgtta
      781 ttcctgttgc aataataata ttttgttatt atcagatagt aaaagctgtt agacttcatg
      841 aactagaaat gttgaagatg gctcaaaaaa tgaatgcttc tcatccagct tccatgaaga
      901 cgggtgctaa aaaagcagat gttcaagctg caaaaatttc tgtgataatt gtctttcttt
      961 atttgatatc ctggactccg tatgcaatta ttgctctgat ggcgcttact ggacgcagag
     1021 atcacctaaa cccttataca gctgaattac ctgtactttt tgctaaaaca tcggctatgt
     1081 ataatccttt tatttacgcg atcaatcatc cgaaatttag aattcaactt gaaaagaaat
     1141 tcccttgttt gatttgttgt tgtccaccca aaccaaagga acgtgacaca acaagtgttg
     1201 cttcggccac gaaaccgagc aaatatcaaa gcgaaaaaga aagttcgatg tctcaacttg
     1261 attctgaaga cgataaacaa aacgtcgaaa ccaacaaggg tcaagctgct agtggtcata
     1321 cgaatgaggc ttacgaaagt gcgggtagaa ttggccgggc ccggtaccca attcgcccta
     1381 tagtgagtcg tattacttcc acataacccc ttggggcctc taaacgggtc ttgaggggtt
     1441 ttttgaggat cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct
     1501 ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa
     1561 caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac
     1621 gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
     1681 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg
     1741 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
     1801 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
     1861 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc
     1921 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
     1981 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
     2041 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
     2101 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat
     2161 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     2221 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     2281 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     2341 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     2401 atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga
     2461 cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc
     2521 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     2581 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     2641 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     2701 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2761 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2821 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2881 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2941 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3001 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3061 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3121 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3181 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     3241 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     3301 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     3361 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     3421 gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta gaaacgcaaa
     3481 aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt atggcgggcg
     3541 tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc ccggcggatt
     3601 tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttc
     3661 cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg ttaacgctag
     3721 catggatgtt ttcccagtca cgacgtt
//
