LOCUS       sequence-208378-        6300 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 6300)
  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 6300)
  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_slit-1
FEATURES             Location/Qualifiers
     source          1..6300
                     /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(3930..3948)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4006..4024)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4029..4045)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4029..4045)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4147..4166)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4158..4967
                     /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(4225..4244)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5114..5702
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5603..5622
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5856..5873
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(6032..6059)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6151..6237)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(6286..6300,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(6300,1..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 cgtaccagat ggactgtttt ccaagctcag caaattacaa
      241 aaattagatc ttcgagataa tcaattacaa tgtatcaatg aaaacacatt caataaactc
      301 actatgctca aatatttact tctagaaggc aacaaaattt tttggatatc agaatcaata
      361 tcgaaattaa aaaatttaaa agaattgaga ataaagaaaa ataaattgac ttgcaactgt
      421 cacctcgcct ggctcaattg gtttatgaaa catcgaccag atatatttaa accagatgtt
      481 ccttcagttg tgtgttcagc accatttaga ttatccgaca tacctgtata cactcttagt
      541 ccatatgaat tccagtgtgc aagtgaagat aaaacgaatt tagaaaacat ttgtcagcaa
      601 aataaaaagg tttgttcgaa acaatgcaac tgttctaatg atggaattgt caaatgtgtt
      661 ggtaaacaac tttcaaatat tcctcaagat ctaccagtaa atgttgttga attagatctc
      721 agtgataata atatcacaat tctccagcaa ggctatttgt ctaaatataa aaccttaaag
      781 aaaataaatt tgaccgggaa tggaatggtc gacattgaac ccggcgcatt ccttggatta
      841 cagcaattga catcactcta tcttaatgca aacaaattga aaaccatcag aaaaaatgct
      901 ttcagtggac tgtcatcact tatgtttcta tttctacacg gcaatgagat tgaatgttta
      961 ccagctgatg ctttcgaggg tgtgaaaaag cttcatatac tacatctatc agaaaacaaa
     1021 ttgacgacct tgaaaagaga aacattcaat ccattgatta aacttagatt tttatatatt
     1081 gtaaataatc ctttaaattg tgactgcaga cttgcatgga ttgtagatta tataaaaggt
     1141 aaagaaaata atgaaccaat tgcttcatgt cactacccga gaaatatgca gggcctagca
     1201 ttgagatcat tgcaaacata cagttttcaa tgtaacaacg aattcgattc gaaatacgaa
     1261 acgcacaatg aagagtgcgt tccaaaagtt agttgtccat taaaatgcac ttgtacatca
     1321 gatattgtga attgttcaaa tactggtttg aacgctcttc cttcaagcat tttcccatca
     1381 actaaaacat tgatcatgtc acataataat ctgaacaaat taacaccgat atattcagtt
     1441 cacgaagcac cgaatctaat gaaattggat ctgtcattca atcaaattga gaaacttgat
     1501 gccgatgtgt tttccagaag cacaaaatta acagaaatac atttgaattc aaacaaactg
     1561 aaatgtatca acaatgaaac tttcaaaaat ctgaatgatt tacgaacttt gcaactgagt
     1621 gacaatgaaa taaactgtat agtggtggat tcattcaaaa acaatctcaa tttaaaatat
     1681 ttaatgctca atagaaatcc attccactgt aactgtaata tgaaatggat gtctgaatgg
     1741 tctagaaatc accacagcac tcttggaaat catcgactgc caacttgtca gtctccaagt
     1801 ttattaaaag gaacaccaat tacgcattta gatgaaaaat attttctttg cgatggaaat
     1861 gcttcaattg atcaatcgtc tgaatcttgc agtgaaatta gttctgctga agcgtgttgt
     1921 tcaatcggaa gtgatgaatg taaacaaatc aaaaaatgtc cattgaaatg taaatgcgaa
     1981 attacaaaag tggattgtag tgatatgcaa ttgaaagaaa ttcctcatga aataccatta
     2041 gatacaactg aactatatct ggacagaaat gatctcaaaa ctcttaatga aacatcattt
     2101 aaaaatttag tttttctaaa aactctagtg ctaagttaca atggcataac ggaactcaat
     2161 aagaatgtgt tgacaccact gaaaaatttg gaaacactag tccttagttt taataaattg
     2221 caatgtattc atcaggatgc tttcaaggac ttaaagaatc taaaagtatt aatactgcaa
     2281 agcaacgatg tctcaacaat tccctttcaa gctttcaatg atttgaacaa tttaaataat
     2341 attgctcttg gacaaaatcc atttcattgt gattgtaata tcaaatggct taatcaattc
     2401 ttccatgaca aatttcttga taatggaata tcattgtgtg caagtccgga aaaaatgaaa
     2461 ttgaaatcaa tttttcattc aaaaccaact gatttcattt gttccgaaga aacagaagat
     2521 gcctacatta cagcaaaatg tgacggatgc atgaaacggc cttgtaaaca taatggaaaa
     2581 tgttcattga tcactcatca taattaccaa tgcgattgtc agtatccttt tcatggaaaa
     2641 aattgtgaaa ataaaataaa tgcctgtttt gggcaaccgt gtcgaaacgg tggaatttgc
     2701 aaaaatattg ataattttgg aaattatgaa tgtgtttgtc cacttgggtt tcgtggtcct
     2761 aaatgtacat taaatattga tgattgcgtg gctcatcaat gtattaatgg agaatgcgtt
     2821 gatctaagaa atagctacag ttgtaaatgt caaaatggat tcagaggaaa atattgcgat
     2881 ataaaatttt catattgtga tgacattaat ccttgtaaaa atgacggaat ttgccaatca
     2941 cttccgaata atgactacaa atgtgtttgc tcaaaaggat gggaaggcaa agaatgtgac
     3001 cagaatattg atgattgtaa atataatact tgcaaacatg aagcaaaatg tgtcgattta
     3061 cataatgatt ataaatgttt atgtagaccg ggttatgtgg gaaagtattg tgaacagcca
     3121 gcatggaagc cgacatcaaa accagtatca cgaagatata ttaatgaaag cgccgtaatg
     3181 atacacagtg gttctgaaaa cgaaattatt tcagaatctg tttgtgctta ccatgaatgc
     3241 ttgaataatg gaatatgtgt aaatggagat caaagtaaaa atcaaaaatc gtattgtaga
     3301 tgtaaaccag gtttctctgg tcatttatgc cagtatttga atagtttata ttttcataaa
     3361 aattcccata ttgaaattac tccaccatca agaggattct taatgccaaa agggaatata
     3421 tctttaaatt ttctgacaaa ttctacttca ggtctattac tttatcaagg attcgatcaa
     3481 acatatttgg tagtaaatct ttatcaaggt catgtgagga taagttataa ccttggtgat
     3541 aaagttgttg gcccggaagg ctattcaatg ggcatggtaa atgatagcaa attacagaag
     3601 cttcacatag aaatagtcgg tgaaaatctg acaatgtatt taaatggccg agttcattgt
     3661 acattgaaat cgaaatatat caattctaat agatatatgg cacttgatac aggaattttt
     3721 attggtggag ctccaaataa aatactgcaa tctgcaatga caaaatatca tattgacact
     3781 gattcagaat ttatcggttg tattgaatcg ttactaatta atcaaaagag aatagatttc
     3841 tcgaaatatt tatctagatc aattaatgta aaaccgggat gtggagatat attgcgggta
     3901 gaattggccg ggcccggtac ccaattcgcc ctatagtgag tcgtattact tccacataac
     3961 cccttggggc ctctaaacgg gtcttgaggg gttttttgag gatcccctat agtgagtcgt
     4021 attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt
     4081 acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca
     4141 gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt
     4201 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag
     4261 gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg
     4321 gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg
     4381 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac
     4441 tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag
     4501 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt
     4561 gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga
     4621 ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac
     4681 aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg
     4741 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg
     4801 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg
     4861 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg
     4921 atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta
     4981 attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac
     5041 caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga
     5101 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5161 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5221 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5281 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5341 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5401 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5461 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5521 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5581 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5641 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5701 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5761 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     5821 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     5881 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     5941 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta
     6001 ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg
     6061 cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg
     6121 cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag cgttcaccga
     6181 caaacaacag ataaaacgaa aggcccagtc ttccgactga gcctttcgtt ttatttgatg
     6241 cctggcagtt ccctactctc gcgttaacgc tagcatggat gttttcccag tcacgacgtt
//
