LOCUS       sequence-208227-        3087 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 3087)
  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 3087)
  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_laminin
FEATURES             Location/Qualifiers
     source          1..3087
                     /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(717..735)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(793..811)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(816..832)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(816..832)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(934..953)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             945..1754
                     /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(1012..1031)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1901..2489
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2390..2409
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2643..2660
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2819..2846)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2938..3024)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3073..3087,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3087,1..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 cgatacacgt gtgagggtat cgctaagcgg agagtgatgg
      241 cgccggatta ttcggtcact gtaacaaagg gatcgatatg caagagaccg ttttacaaca
      301 acgacgcctg gtatcaggaa ttgtgtatac gttgttactg ttccaatgtc acagatgatt
      361 gtaccagtgc gaatggatac gttataaatg cggatccaat agaaagcaaa gtcaaagcgt
      421 ctgatgctgc aattatgaat tacaaaaaca atgtctacta cgaacctgcc cgcgcggtca
      481 cttttgccaa tgaaagcatc ttaatgtacg taataaataa agatgattac aagaaattga
      541 ttccggacga cgaccatttc ttgggcggca gtttcaatat gagagggtca tggattaaca
      601 aatatggatt tccgttaact tataaattga ttatttctgg agaaagcgtt gggcatattt
      661 cgggaccaat agttgtcatg acgggtagaa ttggccgggc ccggtaccca attcgcccta
      721 tagtgagtcg tattacttcc acataacccc ttggggcctc taaacgggtc ttgaggggtt
      781 ttttgaggat cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct
      841 ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa
      901 caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac
      961 gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
     1021 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg
     1081 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
     1141 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
     1201 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc
     1261 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
     1321 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
     1381 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
     1441 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat
     1501 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     1561 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     1621 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     1681 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     1741 atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga
     1801 cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc
     1861 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     1921 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     1981 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     2041 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2101 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2161 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2221 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2281 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2341 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2401 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     2461 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     2521 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     2581 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     2641 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     2701 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     2761 gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta gaaacgcaaa
     2821 aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt atggcgggcg
     2881 tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc ccggcggatt
     2941 tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttc
     3001 cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg ttaacgctag
     3061 catggatgtt ttcccagtca cgacgtt
//
