LOCUS       sequence-208228-        3663 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 3663)
  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 3663)
  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_innexin-10
FEATURES             Location/Qualifiers
     source          1..3663
                     /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(1293..1311)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1369..1387)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1392..1408)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1392..1408)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1510..1529)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1521..2330
                     /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(1588..1607)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2477..3065
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2966..2985
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3219..3236
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3395..3422)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3514..3600)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3649..3663,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3663,1..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 cgacgttacg acctacaaat gggtgaactt cgtcggcatg
      241 gaggactttg ccgacaagct cagcttcaat gtttcgacgt cactgttact attaataacc
      301 atcattataa cagtgaaaag ttacctactg acgtcgatag cctgttacat tcccatacag
      361 ccatcaggta aagatttcga tgactttctg aaaagttatt gttgggtgca cggaaccatt
      421 ccaatgaatg catcgttacc gtggccaaaa gacgaaaatg cttgggaaga actggataga
      481 aatcaaagaa taaattatta tcaatgggtg ccattcgttc tggctctaca ggggattttg
      541 ttctacttgc cacgaattat ctggcaaata atgacctaca atctgtctgg cgcagatctc
      601 gagggtctgg tcgagatggc caactccgcc tgcaatcaaa ccggagaagt caagaataac
      661 acagtcaaac aaattgccga aaatcttgaa aatatgttct tctgtcacag ggaaataaat
      721 ttccacaata gaagtttggc aataaaggct gccaaggcca agaattggct ttgtgccgga
      781 aaacgaagcg gtcgacagtt gataatacag tacatgttca tcaagatgct ctacatgatc
      841 aatgtgttcg gccaattgat tctgatgcag aagttcttgc agttcggcac acacgaccat
      901 tacttcggat atatagtgct aaagaatatc ctgagcggcc gcgaatggaa tgacactcag
      961 attttcccgc gggtcacgta ctgttacgtc aataaaatcc aacatgtcgg catgcagaac
     1021 gattatctgg cccagtgtgc tctgccaatt aatatgctca acgagaaggt gtacatcttt
     1081 ctttggttct ggactgtgat agtcgcggtg ttcactgtca ccagtgtcgt gcaatggatc
     1141 ttccggattg cgttgacaaa caatgaagtg ggcttcttga agaaattctt acgggttcgt
     1201 gaacagtacc aaaaagagga tttgccgagt ttaaatcgct tctgtgaggg atttctacgg
     1261 gtagaattgg ccgggcccgg tacccaattc gccctatagt gagtcgtatt acttccacat
     1321 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gaggatcccc tatagtgagt
     1381 cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat
     1441 gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa
     1501 acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa
     1561 attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat
     1621 caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac
     1681 atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga
     1741 cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt
     1801 tactcaccac tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt
     1861 caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg
     1921 tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa
     1981 tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg
     2041 aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc
     2101 atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg
     2161 atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc
     2221 tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc
     2281 ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg
     2341 ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat
     2401 gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa
     2461 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     2521 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     2581 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     2641 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     2701 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     2761 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     2821 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     2881 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     2941 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3001 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     3061 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     3121 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     3181 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     3241 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     3301 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatacgc
     3361 gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt
     3421 ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg
     3481 ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga gagcgttcac
     3541 cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc gttttatttg
     3601 atgcctggca gttccctact ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac
     3661 gtt
//
