LOCUS       sequence-208375-        4605 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 4605)
  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 4605)
  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_laminin
FEATURES             Location/Qualifiers
     source          1..4605
                     /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(2235..2253)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2311..2329)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2334..2350)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2334..2350)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2452..2471)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2463..3272
                     /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(2530..2549)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3419..4007
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3908..3927
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4161..4178
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4337..4364)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4456..4542)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4591..4605,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4605,1..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 cgcctgaacc tggaaacttt tatttatatc attcaggagc
      241 taagaaggaa ttacattgtg aaattgagcg agtgaataac cagtggataa tgaaggcaaa
      301 agatgtaata tggacattta ataacaaaaa aattgaaact attgcaaact cggttttaaa
      361 aagagtagat agattagata caaattcagt cagtatactt gtattgacag atattgacga
      421 aaaacatgaa ggaaattata actgttttca aggatcaact gtacttagtg caaaattatt
      481 aatggaacgt agtaacattc tccttgaact agctccacca ctgcaagaag tggcagacaa
      541 tgtaaatcaa ggtgctgaat tacactgtcg cattgataca cagaatgtta attcaagttc
      601 aattatttgg aaatttattc cggaaaaatc attaattgaa acagagttac caatcgaagc
      661 taaaattgaa agaccttcat taacatcttc aacatccttt attataatta acaatgtttt
      721 gaaaaggcac gaaggcacat atgtttgcca agtcctgaac accaaagtat tcggaaaaat
      781 tattgtgaga acaaaacctg tgttgactgt taccccaaaa tcgataacta taaaacctgg
      841 tagtactgct agatttcttt gtcaattaaa tggacttgaa ggcaaagtaa tttggcgaag
      901 aacggataga aaaagtttga ctgtgaatag agaaaataga ttaggaggtg atgcatcaag
      961 tttattggaa atcaaagatg ttacacgaga tgatcacgga accatttatg aatgtgtttt
     1021 taataaatta acagaatttg taactctaaa tgtcgaagaa gaatgtgaat taaatcaacg
     1081 taaatgtaaa aatgaaattt gtattgatga gaacaaattt tgtgatggta aagatgactg
     1141 tggcgataat tcagatgaag accgtcaaag atgcaatgaa tgtaatccga attcaatcca
     1201 ctgtgacttt cttaataatt caaatccaaa taaacgatgt gttataaagc attggcaatg
     1261 cgacggaatt gatgactgtg gaaataattt cgatgaacta aattgtcccg ttaaaaatca
     1321 aaacaggtgt attggaacac actttctttg taacaacaat actttgattc ctcgttcgta
     1381 ttggtgtgat ggtgaaccag actgtttatc tatggaagac gaaatagatt gcagcgaccc
     1441 acaagttata gagcctgtga agcaaatgca aatttcaata tttacaaact caactttaat
     1501 cttgacttgt aaaatgtcag gacgtccagt tccaactatt tcatggagat ttaactggga
     1561 tcatttgcct gtaaatttaa atcataatat tacaacaaca acttccaatt gtagcatgat
     1621 tacaagcaca ttactaataa aagatataag tacagaccac agtggtgtat attcttgtga
     1681 aggtatttca agaagaagag taatggcacc agacttttct gtatcagttt caagattaat
     1741 atgccagaaa ccatattata ataatgatgc ttggtatcca gaaatgtgtc tcaggtgtta
     1801 ttgttcaaat gttacagatg actgtacaag tgcgacagga tatatcacaa aacctgaacc
     1861 aattgaaagt ggaattaaac catctgatgc agcaatagtt aatttcaaaa ctcttgaata
     1921 ttataagcca gctagagaaa taacatttgc taatatggaa aaaatgaatt actttattaa
     1981 tgaaacatat tatcaacgct tagttcctaa tcaggattac tattttggag gtgcatttaa
     2041 tatgggtggc tcatggttaa aacgatatgg ctaccctatg acttataagt taaagttatc
     2101 tggagaaagc aagacctata ttcctaatcc aatggttatg ataaaaggag aagttgcttc
     2161 aatatattat tgtggcgtta aatatcggtt gccagtttac gggtagaatt ggccgggccc
     2221 ggtacccaat tcgccctata gtgagtcgta ttacttccac ataacccctt ggggcctcta
     2281 aacgggtctt gaggggtttt ttgaggatcc cctatagtga gtcgtattac atggtcatag
     2341 ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata
     2401 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     2461 ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg
     2521 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     2581 gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     2641 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     2701 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     2761 ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     2821 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     2881 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     2941 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     3001 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     3061 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     3121 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     3181 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     3241 agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact
     3301 ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg
     3361 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3421 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3481 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3541 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     3601 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3661 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3721 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3781 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3841 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3901 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3961 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4021 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     4081 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     4141 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     4201 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     4261 ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag
     4321 agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct
     4381 ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa
     4441 atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa
     4501 acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta
     4561 ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgtt
//
