LOCUS       sequence-209324-        5558 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 5558)
  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 5558)
  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: Pgr_laminin
FEATURES             Location/Qualifiers
     source          1..5558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1..17
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        22..40
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        98..116
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3136..3152)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3136..3152)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(3210..3228)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      3233..3280
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3287..3304)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3287..3303)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3296..3318)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      3367..3453
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3545..3572
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3731..3748)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3902..4490)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3982..4001)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4637..5446)
                     /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     5360..5379
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5438..5457
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 caggaaacag ctatgaccat gtaatacgac tcactatagg ggatcctcaa aaaacccctc
       61 aagacccgtt tagaggcccc aaggggttat gtggaagtaa tacgactcac tatagggcga
      121 attgggtacc gggcccggcc aattctaccc gatgaccaac actctctcca gatattataa
      181 ttttataggt gagtggatat ccatatcgat taatccatga tcctcgcata ttgaaacttc
      241 ctcctagaaa taaatcgtcg gaagttacaa atttctcata attggttgta ttgattgaat
      301 acgttttgga tacatcgtcg acaaaactga ccattctagc gggttcgaag tattcgtttg
      361 tttgataatt cattagggcg gcatctgaag acctgacttt actctcaact agatcaggat
      421 tagtaatata tccattggca ctagtgcaat cgtcagttac atttgagcaa taacatctca
      481 gacagaattc ctggtcccac gcgtcattgt taaagtatgg tcttttgcaa atagacccct
      541 ttgtaacggt taccgagaaa tctggagcca ttatacgtcg tttctcaata ccctcgcaag
      601 tgtaaatccc gctgtcctct ggcgagatgt tcgtaatggt tagcgtgctc gtaatgagag
      661 tacagttagt gacgacctcc gtgacgttgt gatccatagt gtcaggcaga tagccccaat
      721 tgtatcgcca agtgattgtg ggcataggtc gtccagaaat ttggcacaca agggtcaagg
      781 tgctgttttc gattaccgac aagtgactta acttggctgg ctctatcacc tgtggaggac
      841 tgcatccttc ttcgtcttcc atggacttgc aatcggaatc cccgtcacac cagtacgctc
      901 tagggatcag ggttccattg tcgcacgagt agtgtgagcc taggcacttg tctttgttct
      961 tgtctggaca atggagttca tcgaaattgt ttccacagtc atctatgcca tcgcattgcc
     1021 aatgtctcac tacacaccgt ttaataggcg aagaattgtt caggaaatca caatgtattg
     1081 agtttggtgt gcattctcta catcgcttcg gatcttcatc cgagttgtcc ccacagtcat
     1141 ttttaccgtc acagaacttt ccttcgtcta tgcaagtctc gttttggcat tttctttgat
     1201 tcaaagcaca ttcgtctttg acccataatg tgacaatttc agtcaatgaa ttgaaattgc
     1261 attcataaat ggttccatca tcggctttgg taacttcttt gatttccaat aaggaatttg
     1321 tactccctcc gagtctgttt tctttattca gtataaaact ctttttgtct tttcgtcgcc
     1381 atgttacttt tccatcgata gcagtcaatt ggcaaaggaa tctcgcagtt ccaccgggcg
     1441 aaacagtgat ttctttcgga tttactgtca gcatttgttt tgtcttcaca atcaatattc
     1501 cagtgacttg ccgacctaac actttacaaa cataaaaacc ctcgtgtttc tttaaagaat
     1561 ttttaatagt cacaaacgac gtcgccgacg tttgatgagg cttctcgaca atagcctctg
     1621 gtggtagatc gtattctatg gtggaattga atttcataaa cttccattca atgaaactcg
     1681 agttgtatcc aggggtgtct attctacaat ggaactctgc cacgtgattg gcattgtgca
     1741 acaccgttag acgaggagga gacaattcta tcatcacctc gctttgttcc attataagaa
     1801 tagacgcaaa tgtcaagggg ccttggaaac atttgtatac accttggttg tcttcgccca
     1861 tgtctttaag gaccaaaata ctcaccgatc tcgtgtctag gcgtccaaat cgattgatat
     1921 taccaaaagc agacaatgga ttttccacgt caatctttct tccattgaat gtccagataa
     1981 tatcctttga tctgaatttc cattctccgt cttctctgac aatctcacaa tgtaattctt
     2041 ttgttttctc gggatgaaaa ttgtaataaa gtccgggatc cggagtcatc tctattttag
     2101 attctgtgcc cgccttcggt gtcagattga taatacgaga ctgaatcatc acaccgttgc
     2161 gatggatttc acaactatac ttgccgtaga cttctttttg aaagtttttg atgaaatatg
     2221 tgttaccgcg atgaattatt ttgtcatcta atctccagat aacctcggtt ttattcccca
     2281 agtccggtga tggaatacag ttaactttca tctcagctcc gtatgccgct cggccctcca
     2341 ctgacatctc cacctctcta tttttccact cagaaatgaa cactttaccg actttgttca
     2401 cggtaaaatc tccgttctta ttgattctat aataaagagg taagtctgat ttactggcgg
     2461 cgtacgaata gaaataagca tctcgtccgt gttctagaag aatggttcct ggtggttgtt
     2521 cccaggttac ggtctcagaa attccggcaa catttcctgg tcggtagcgt atctctttgg
     2581 cctttggaat tatattttcg agatacatga catcttcgtc gatagttttt tctcgctttg
     2641 ttggtctttg aggaagtaat attacttttg tcacttgtaa cactgatttg gaggtcatac
     2701 caatagcaaa gtaataggtc ccgaaatcaa attcattcaa attatcaatt ttaagctttc
     2761 tgccagtgaa tatatggaat tggggtggcc ggtaccaagt caccatgtca ttggttgaag
     2821 cctctgaatc caattcaacg gatccagatt tttctatgac gtatttagtc tttgaataat
     2881 cgtagacaaa cttaacagga ctttccattt ctttgctgtc gataatattt agagtagcgt
     2941 ttctcgttga tgaatcttta gaaataattg aacactgata ttctccaagg tcggactttg
     3001 ttttaatttt taaaataata ccatgaggct tccgtttctg tattagccct ttgattcctt
     3061 tctccacttt atagtacatg accttatcct catttgacct gcgggatggt aatgccgcta
     3121 gccatggaac cggtggatcc actagttcta gagcggccgc caccgcggtg gagctcgaat
     3181 tcatcgatga tatcagatct gccggtctcc ctatagtgag tcgtattact tcctagcata
     3241 accccttggg gcctctaaac gggtcttgag gggttttttg cttaagactg gccgtcgttt
     3301 tacaacgtcg tgactgggaa aacatccatg ctagcgttaa cgcgagagta gggaactgcc
     3361 aggcatcaaa taaaacgaaa ggctcagtcg gaagactggg cctttcgttt tatctgttgt
     3421 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgtg
     3481 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaact
     3541 aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct tcctggctag
     3601 cggtacgcgt attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     3661 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     3721 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3781 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3841 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3901 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3961 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4021 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4081 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4141 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4201 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4261 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4321 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     4381 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4441 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4501 tgatcttttc tacggggtct gacgctcagt ggaacgacgc gtaactcacg ttaagggatt
     4561 ttggtcatga gcttgcgccg tcccgtcaag tcagcgtaat gctctgccag tgttacaacc
     4621 aattaaccaa ttctgattag aaaaactcat cgagcatcaa atgaaactgc aatttattca
     4681 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
     4741 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
     4801 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
     4861 caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga
     4921 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
     4981 tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat
     5041 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
     5101 cacctgaatc aggatattct tctaatacct ggaatgctgt ttttccgggg atcgcagtgg
     5161 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
     5221 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
     5281 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaagcga tagattgtcg
     5341 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
     5401 tggaatttaa tcgcggcctc gacgtttccc gttgaatatg gctcataaca ccccttgtat
     5461 tactgtttat gtaagcagac agttttattg ttcatgatga tatattttta tcttgtgcaa
     5521 tgtaacatca gagattttga gacacgggcc agagctgc
//
