LOCUS       sequence-209326-        3908 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 3908)
  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 3908)
  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_collagen
FEATURES             Location/Qualifiers
     source          1..3908
                     /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(1538..1556)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1614..1632)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1637..1653)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1637..1653)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1755..1774)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1766..2575
                     /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(1833..1852)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2722..3310
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3211..3230
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3464..3481
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3640..3667)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3759..3845)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3894..3908,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3908,1..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 cgaggagatc caggagagac tggagcaccg gggcaagatg
      241 gggaggaagg taaagaagga gctcctggag cccctgggaa acctggtctt ttaggtccac
      301 aagggttgga aggaaatatg ggatctcctg gtccctctgg agaagcggga gacccagggg
      361 caacaggtgc ccctggtccc gttggagcac caggtcgaga tggtagccta ggtttacctg
      421 gaagacaagg aaaaccaggt gagccaggta aagaaggcgc cgtaggaaaa gtaggtagtc
      481 ctggagtcat tggtgctcca ggaaataatg gagctgatgg gcgaaaagga cctttaggtc
      541 caagggggcc acctggtgac aaaggcaagc caggaggacc aggtactgag ggaaaaccgg
      601 gatctgatgg ttcacccgga gaccctggat ttatgggttc gccaggtcca tctggaccac
      661 aaggaagcct tggtaaacct gggaaacctg gtccagctgg tagcgatggt ttaccaggag
      721 actatggaga aatgggagtt ccgggtccag caggaccaga tggagcacct ggaaattctg
      781 gaactgcggg agcacgaggg ccatctggat taccgggagg caaaggaaaa ccaggacctg
      841 ctgggaaggc tggaaagcca ggaaatggag gttttgctgg cattcctggt gctaggggtg
      901 cgactggaac tccgggacca gctggagatg ctggtccttt aggtgctcca gggtctaatg
      961 gtgctattgg agaaactggt ctggctggtg ataaaggtgt atcaggagca aatggaaaac
     1021 caggacaagc tggacgacca ggtccaaggg gccctgcagg cacaaaagga cccaaaggaa
     1081 gtcctggttt tgatggacca actggagctg atggtgacag gggtgctatt ggctatcctg
     1141 gcccaaatgg tcccccaggt ccaagtggtc cagcaggact cactggtcct actggagtga
     1201 ttggaccacg aggtgatcct ggtgttctag gacctgttgg agtcgctggt ttacctggaa
     1261 gacagggtga aagtggaccc ataggacaaa gaggtccaat tggcccacca ggccctcctg
     1321 gacccccagg tggagttatg gcaatgcaga tgagatcacc aacaaaagga attatgtatg
     1381 gtgatgattc gaagaccgct aaattacttg gtggtaatgc tataaaaaat ccccaaggaa
     1441 cgaaggatat tccggcgaaa acatgcaagc atttgagcgt cagttatccc aataaacccg
     1501 accgggtaga attggccggg cccggtaccc aattcgccct atagtgagtc gtattacttc
     1561 cacataaccc cttggggcct ctaaacgggt cttgaggggt tttttgagga tcccctatag
     1621 tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct
     1681 ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta
     1741 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg
     1801 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     1861 gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct
     1921 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     1981 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     2041 atggttactc accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc
     2101 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     2161 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     2221 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     2281 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
     2341 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     2401 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     2461 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     2521 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga
     2581 attggttaat tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag
     2641 ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt
     2701 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2761 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2821 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     2881 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2941 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3001 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3061 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3121 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3181 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3241 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3301 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3361 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     3421 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     3481 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     3541 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     3601 tacgcgtacc gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg
     3661 gccttctgct tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg
     3721 ggccgttgct tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg
     3781 ttcaccgaca aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt
     3841 atttgatgcc tggcagttcc ctactctcgc gttaacgcta gcatggatgt tttcccagtc
     3901 acgacgtt
//
