LOCUS       sequence-208376-        3610 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 3610)
  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 3610)
  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_collagen
FEATURES             Location/Qualifiers
     source          1..3610
                     /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(1240..1258)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1316..1334)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1339..1355)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1339..1355)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1457..1476)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1468..2277
                     /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(1535..1554)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2424..3012
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2913..2932
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3166..3183
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3342..3369)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3461..3547)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3596..3610,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3610,1..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 cgaggaaaac aaggacctga aggattaagt ggcccagaag
      241 gtgcttctgg tccacccgga gatgttggtg atcctggaaa atctggccca ggtgggccgc
      301 gcggtggtcc tggcagacaa ggtagtccag gaggtccagg gaaacaagga aaagctggta
      361 ttcaaggtaa acctgggcct gctggaaaac ctggacctct tggtcctcaa ggagttacag
      421 gtttagatgg atcaggtggt cgaaaaggtc ctttaggttt aagaggtcca gtaggtccaa
      481 aaggtaaacc cggagctcct ggtactgatg gaaaagttgg tacatctggt cctgacggct
      541 atcctggata tttgggtcca ccaggaccct cgggtccaca ggggataaca ggaaaaccag
      601 gaaaaaaagg tcctccagga aatcgtggtc caataggaga taatggagaa atgggcaaac
      661 ctggtccggt cggtgtagtt ggagatcctg gaattactgg tccagtaggt ccacgaggtg
      721 accctggacc tcctggtagt aaaggagctg caggcagtcc aggtaaagat ggaacaagtg
      781 gtgggaaagg atatgatggc caaggaggaa ctagcggaaa atcaggaaag ttaggaactc
      841 ctggtgataa aggtgcagat ggtaagcctg gaccaggcgg tgtacttgga gatgttggtt
      901 tgagaggagc aaaaggaaaa aaaggaaaag atggagtacc aggacgtgaa ggaagaccgg
      961 gtcctagagg tgcaaaaggt gtaaaaggac caaaaggagt ccctggatat ccaggattaa
     1021 tagggataga aggagatcca ggaccaccag gttttcctgg tccagatggg cctgttggac
     1081 ctaagggtcc tgatggaata caaggtccag ttggaataac tggtccccgt ggtgatcctg
     1141 gaataccagg agctattggt ccaactggtc tttcgggtaa gaaaggttca aatggtcctt
     1201 taggcgggta gaattggccg ggcccggtac ccaattcgcc ctatagtgag tcgtattact
     1261 tccacataac cccttggggc ctctaaacgg gtcttgaggg gttttttgag gatcccctat
     1321 agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat
     1381 ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct
     1441 tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg
     1501 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
     1561 gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt
     1621 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
     1681 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     1741 gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat
     1801 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
     1861 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
     1921 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
     1981 cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc
     2041 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     2101 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     2161 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     2221 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca
     2281 gaattggtta attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca
     2341 agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
     2401 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     2461 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     2521 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     2581 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     2641 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     2701 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     2761 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     2821 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     2881 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     2941 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3001 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     3061 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     3121 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     3181 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     3241 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     3301 aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga
     3361 tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc
     3421 cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag
     3481 cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga gcctttcgtt
     3541 ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat gttttcccag
     3601 tcacgacgtt
//
