LOCUS       sequence-208383-        4417 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 4417)
  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 4417)
  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_laminin
FEATURES             Location/Qualifiers
     source          1..4417
                     /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(2047..2065)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2123..2141)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2146..2162)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2146..2162)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2264..2283)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2275..3084
                     /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(2342..2361)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3231..3819
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3720..3739
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3973..3990
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4149..4176)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4268..4354)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4403..4417,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4417,1..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 cgaatacaac ggtacttgca gcaaaacttt tgatggagag
      241 aagcgatgta ttaattgagg ttgctccatc tgaacaaact gtattcgata atgttaacca
      301 agaatccgaa ttgcattgtc gaatagatac tgctaacatt aattcaagct tcatacaatg
      361 gtcatttatt ccagaaaatt ccaaaacgga agtagaactt ccacctgaag ctaaggtttc
      421 tcgaccagca ttgacctctt caacttcatt tgttgtaatt gaaaacacct tgaaaagaca
      481 tgaaggtaca tatgtctgca gagttcttgg taaaaaggtt tttggccgag tgattgttaa
      541 ggcaaaacca gtattgaaag tttcccccaa aacaataacc attaaacctg gaggtgatgc
      601 gagatttctt tgtcaactaa tgggagcaga tggaaaagtt atttggagga gatttgacaa
      661 aaagagttta gaagtcaaca aagaaaacag aattggtgga agtactcaga gtatattaga
      721 aataaaacaa gcaacgaaaa atgatcacgg aactatttat gaatgtgttt ttaataaact
      781 aattgagcaa gtgacgctct ttgttgaaga agaatgtgct ttgaatcaaa gaaaatgtaa
      841 aaatgatctc tgtatagatg aaaataagtt ttgtgacggt aaaaatgact gtggagataa
      901 ttctgatgaa gaccgaacga gatgcaatga atgtagccca aacgctattc actgcgactt
      961 tctgaaaggt gcagatccga ccaaacgttg tgttataaag cattggcagt gtgacggcat
     1021 tgatgactgt ggaaataact ttgatgagct taattgtcca gttaagaaca cggacaaatg
     1081 cattggcact cattttctct gcgatgacga tgttttgatt cccagatcat attggtgtga
     1141 tggagattct gattgcaaat caatggaaga tgagaagagt tgcagtcatc ctcaagtagt
     1201 agagcctgaa aaacagatgc aaatatcagt ttttagcggt tcaactttaa ctctaacttg
     1261 tgaaatgtca ggccgtccag taccctcaat cacatggaga tacaattggg gacatttatc
     1321 tgagaatctt ttgtataata taacgacgaa gatatcaaat tgtagtgtaa taacaagcac
     1381 tttaactgtt cacaacatca ccatggatca aagtggagtt tattcttgcg aaggaatagc
     1441 aagacgtcga gtgatggcac ccgacttcat tgttgtagtt tcaaagggct caatttgtca
     1501 aaaaccattt tataacaatg atgcgtggga tacgtcaatg tgcatcaaat gttattgctc
     1561 aaatctgacc gatgattgca ccagcgctac tggctacgta aataatccgc aaccagttga
     1621 aagtggtata aagccttcag atgctgccat tgtcaattac aaaacaactg aatattatga
     1681 accggcgcga gaagtcacat ttgcaaatgc aaatttaaag aaatactata ttgataaaaa
     1741 ttcctatatt aaattcatac cgactgccga ttattatttt gggggagttt tcaatgtgga
     1801 aggatcttgg ttgaacagat atggctttcc tctatcgtac caactaattt tatcaggtga
     1861 aagctacaaa tatatacctg gtccacttat cgtaataaag ggaggctctg actatgtata
     1921 ttattgtggc atccaaaatg atttccgtct cccagtttat gcatcggaag atagttatga
     1981 aaataaaatc agagtggtct tatgggaaca acgggtagaa ttggccgggc ccggtaccca
     2041 attcgcccta tagtgagtcg tattacttcc acataacccc ttggggcctc taaacgggtc
     2101 ttgaggggtt ttttgaggat cccctatagt gagtcgtatt acatggtcat agctgtttcc
     2161 tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata
     2221 tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
     2281 catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat
     2341 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat
     2401 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     2461 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     2521 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa
     2581 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     2641 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     2701 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     2761 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
     2821 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     2881 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     2941 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     3001 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     3061 ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc
     3121 attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac
     3181 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3241 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3301 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3361 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     3421 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3481 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3541 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3601 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3661 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3721 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3781 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3841 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3901 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3961 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     4021 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     4081 aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta
     4141 gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt
     4201 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc
     4261 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
     4321 cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg
     4381 ttaacgctag catggatgtt ttcccagtca cgacgtt
//
