LOCUS       sequence-208373-        3791 bp    DNA     circular SYN 15-APR-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 3791)
  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 3791)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_opsin
FEATURES             Location/Qualifiers
     source          1..3791
                     /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(1421..1439)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1497..1515)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1520..1536)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1520..1536)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1638..1657)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1649..2458
                     /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(1716..1735)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2605..3193
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3094..3113
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3347..3364
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3523..3550)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3642..3728)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3777..3791,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3791,1..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 cgctttagtt ggtttgggac agaaaatcct tgaaaattca
      241 acagctgata atgcctcact cagtaaatca ccatggcatt gggatcctga atttgaatcg
      301 attgttcatc catattggag aacttttgat atggttcctg aagtgtatca ttatttagtt
      361 ggagtataca tctctatagt tggaatagcg ggtgttctag gaaatctact cgttctttat
      421 atatttgcta gtgcaaaaag tttgagaaca ccaccgaata tgtttattat gagtttagct
      481 attggagatt tgacattctc agctgtcaat ggattcccat tattgacaat atcgagtttt
      541 aatcatcgat gggcatgggg aaaattaaca tgtgaaattt atggatttat tggaggactt
      601 tttggtttta tatcgattaa tactatggct ctcatttctt tggacagata ttttgtaata
      661 gctcaaccgt ttcaaaccat gaaatctttg actattaaaa gagcaattat tatgttggtt
      721 ttcgtttggc tttattcttt gatctggtca actccaccat tttttggtta tggaaattat
      781 gtaccagaag gatttcaaac ttcttgtact tttgactatt tgacacaatc caaaggaaac
      841 atcattttta atattggaat gtatatcgga aattttgtga ttccagttgg aacaattatt
      901 ttttgctatt atcaaatcgt aaaagccgtt agagttcacg aattagaaat gttgaaaatg
      961 gcccagaaaa tgaatgcttc acatccgaca tcaatgaaaa ctggagccaa aaaagcagat
     1021 gttcaagctg ccaaaatttc cataataata gtgtttttat atttaatttc atggactcct
     1081 tatgcgataa ttgcattgat ggctctcact ggtagacggg atcatttgaa tccatacact
     1141 gctgaattac cggtgctctt tgcaaaaaca tcagctatgt acaatccatt tatttatgct
     1201 atcaatcatc cgaaatttag aatccaattg gagaaaaaat tcccatgctt aatatgctgt
     1261 tgtccaccta aaccaaaaga gagagataca acgagtgctg catcagcaac aaaaccaagt
     1321 aaatatcaaa gtgagaagga aagttctatg tcacaattag attctgagga cgataaacaa
     1381 gcaaccgggt agaattggcc gggcccggta cccaattcgc cctatagtga gtcgtattac
     1441 ttccacataa ccccttgggg cctctaaacg ggtcttgagg ggttttttga ggatccccta
     1501 tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa
     1561 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     1621 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc
     1681 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     1741 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt
     1801 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     1861 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     1921 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata
     1981 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     2041 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     2101 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     2161 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt
     2221 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     2281 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     2341 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     2401 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     2461 agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc
     2521 aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     2581 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2641 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2701 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     2761 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2821 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2881 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     2941 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3001 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3061 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3121 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3181 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3241 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3301 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3361 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     3421 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     3481 taatacgcgt accgctagcc aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg
     3541 atggccttct gcttagtttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct
     3601 ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga
     3661 gcgttcaccg acaaacaaca gataaaacga aaggcccagt cttccgactg agcctttcgt
     3721 tttatttgat gcctggcagt tccctactct cgcgttaacg ctagcatgga tgttttccca
     3781 gtcacgacgt t
//
