LOCUS       sequence-208226-        2903 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 2903)
  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 2903)
  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_opsin
FEATURES             Location/Qualifiers
     source          1..2903
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(131..158)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(250..336)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     385..407
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     399..416
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     400..416
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(423..470)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        475..493
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     551..567
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     551..567
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(932..950)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1008..1026)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1031..1047)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1031..1047)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1149..1168)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1160..1969
                     /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(1227..1246)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2116..2704
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2605..2624
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2858..2875
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
       61 ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac cgctagccag gaagagtttg
      121 tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttagtttgat gcctggcagt
      181 ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc
      241 tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa
      301 ggcccagtct tccgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg
      361 cgttaacgct agcatggatg ttttcccagt cacgacgttg taaaacgacg gccagtctta
      421 agcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctag gaagtaatac
      481 gactcactat agggagaccg gcagatctga tatcatcgat gaattcgagc tccaccgcgg
      541 tggcggccgc tctagaacta gtggatccac cggttccatg gctagcggca ttaccatccc
      601 gtttcgttag atcgcttctt cgtcattgcg cagccgttcc agacgatgcg agctctgtcg
      661 atgaagaaag cgatcattat gctagtcggt gtctggatct attcgttcat ttgggcctcg
      721 cctccattct ggggatacgg caactatgtg ccagaaggat tccaaacatc gtgcactttt
      781 gattatttga cccagagcaa agggaatatt atattcaata ttggaatgta tatttgcaac
      841 ttcgtgttcc cagtcggaat aataatattt tgttattatc aaatcgtcaa atccgtcggg
      901 tagaattggc cgggcccggt acccaattcg ccctatagtg agtcgtatta cttccacata
      961 accccttggg gcctctaaac gggtcttgag gggttttttg aggatcccct atagtgagtc
     1021 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     1081 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     1141 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     1201 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     1261 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     1321 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     1381 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     1441 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     1501 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     1561 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     1621 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     1681 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     1741 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     1801 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     1861 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     1921 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     1981 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     2041 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     2101 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2161 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2221 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     2281 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2341 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2401 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2461 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2521 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2581 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2641 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2701 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2761 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2821 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     2881 ggaagagcgc ccaatacgca aac
//
