LOCUS       sequence-208801-        3686 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 3686)
  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 3686)
  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: Gsp_innexin-10
FEATURES             Location/Qualifiers
     source          1..3686
                     /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(1316..1334)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1392..1410)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1415..1431)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1415..1431)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1533..1552)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1544..2353
                     /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(1611..1630)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2500..3088
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2989..3008
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3242..3259
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3418..3445)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3537..3623)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3672..3686,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3686,1..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 cggaagggaa gactttgcag acaaattgag ttttagtatt
      241 tctacatcaa ttctattgtt gttaacatta gtaataactg taaaaagtta tttattaaat
      301 tctattgctt gttatattcc aattcatccc agtggaaaag attttgatga ttatctaaaa
      361 agttattgtt gggttcatgg aacaattcca attcaaggtc gtttaccaca agatattaca
      421 gattgggaaa atatggataa atacaataga ataaattatt atcaatgggt tccattcgta
      481 ttggcaatac agggtatcct attctattta ccgagaatta tttggcaaat gattacgtat
      541 aatatgtcag gaactgatct tgaaggtctt attgaaatgg cgaattctgt cagcaataaa
      601 acaggagatg caaaaagaga aactgttaat caagttgctt tgaacttaga aaatctattg
      661 tttacgcatc gtaaaattaa tctacatagc gtaccgtttg ctgcaaaatt ttccgaagtg
      721 cgatcatggt tctgttctgg taaaagaagt ggtagacagt taactgctca atatatgttt
      781 ataaaggttc tttatgtcat taacgcgttc ggtcaattat cccttatgca gtactttcta
      841 aattttggac ataaagatta tagttttggt tatttcgttc taaataatat aataaatggt
      901 aaagaatgga atgaaacatt gatttttcca agagtaacct actgctatgt aaagaatatt
      961 cagcatattg gtatgaaaaa cagttatttg gcacaatgtg ctttaccaat taatatgtta
     1021 aatgagaaaa tctatatttt tctttggttt tggacggttc tagttgtgtt ttttacttgc
     1081 attagttttt ttcaatggat ctatcgatta accaaatcca attcacaggt tcgatttttg
     1141 aagaaatttc tgagaatccg tgaacaatat acagatgaag atgatgtagt tttaaaatca
     1201 ttctgtaaag gatttctaag aaaagatggt gtttttattt tacgaatgtt ttcaataaat
     1261 gcgggtgatt tgattacaac cgggtagaat tggccgggcc cggtacccaa ttcgccctat
     1321 agtgagtcgt attacttcca cataacccct tggggcctct aaacgggtct tgaggggttt
     1381 tttgaggatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     1441 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
     1501 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     1561 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     1621 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
     1681 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     1741 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     1801 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     1861 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     1921 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     1981 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     2041 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     2101 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     2161 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     2221 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     2281 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     2341 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     2401 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     2461 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     2521 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     2581 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     2641 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     2701 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     2761 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     2821 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     2881 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     2941 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3001 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3061 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3121 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     3181 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     3241 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     3301 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     3361 tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa
     3421 aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt
     3481 cctgcccgcc accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt
     3541 gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc
     3601 gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc
     3661 atggatgttt tcccagtcac gacgtt
//
