LOCUS       sequence-360128-        3053 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dsRNA and riboprobe synthesis for Schmidtea mediterranea Slit-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3053)
  AUTHORS   Wolff A, Wagner C, Wolf J, Lobo D
  TITLE     In situ probe and inhibitory RNA synthesis using streamlined gene 
            cloning with Gibson assembly.
  JOURNAL   STAR Protoc. 2022 Jun 14;3(3):101458. doi: 
            10.1016/j.xpro.2022.101458. eCollection 2022 Sep 16.
  PUBMED    35733605
REFERENCE   2  (bases 1 to 3053)
  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: "STAR 
            Protoc."; date: "2022-06-14"; volume: "3(3):101458. doi"; pages: " 
            10.1016/j.xpro.2022.101458. eCollection 2022 Sep 16"
COMMENT     Sequence Label: pAW2-Smed-Slit-1
FEATURES             Location/Qualifiers
     source          1..3053
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(55..102)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     104..120
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        121..139
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        141..159
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(1024..1042)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1025..1042)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(1044..1062)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1063..1079)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1063..1079)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      1081..1128
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1135..1152)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1306..1894)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1386..1405)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2065..2856,2857..2925))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2688..2707
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2926..3030)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcaaaaa
       61 acccctcaag acccgtttag aggccccaag gggttatgct agggtaaaac gacggccagt
      121 taatacgact cactataggg aattaaccct cactaaaggg aggacttgcc agtcaccaat
      181 catattgaaa gatactccca ttacacattt ggatgacaag tattttctat gtgacggtaa
      241 tgcgtctgtt taccattcgg ttgattcgtg tcatgaaaca aattccgtga agacctgctg
      301 ctcaattgga aatgaagagt gcactaaatc caaaaaatgt ccttcgaaat gcaaatgcga
      361 atcaaccaaa gtggattgta gtgatttaca gttaaaagaa attccaaatg aaattccaaa
      421 agatactaca gaattatatc tcgaccgaaa ccatctgcaa agtctaaatg aaacatcttt
      481 caagaattta ttaaatttaa aaactttagt tctcagctat aatggtatta ctgaattgaa
      541 caagaatgtt ttgactccat tgaaaagtct ggaaacctta gtgttgagct tcaacaaatt
      601 acaatgtatt catcaggatg ccttcaaaaa tctacacaat ttaaaagttc taatactcca
      661 aagtaatgat gtttctacga ttccttacca agcttttaac gacctgaaaa atctcaataa
      721 catagctctg ggacaaaatc catttcattg tgactgcaat ataaagtggt taaaccagtt
      781 ctttttagat agattcctgg ataatggaat ctcattgtgt gctagtcctg aaaaaatgaa
      841 attaaagtct atttaccatt ccaaaccaac tgatttcatc tgttctgaag aaaccgaaga
      901 tgcttatatt actgctaagt gtgacagttg catgaagaga ccatgcaaac ataacggaaa
      961 atgttccttg ataacccatc ataattatca atgtgattgt ccttatccat ttcatgggaa
     1021 aattctatag tgtcacctaa atccctatag tgagtcgtat tagtcatagc tgtttcctgt
     1081 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgcg cttcctcgct
     1141 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     1201 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     1261 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     1321 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1381 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1441 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     1501 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     1561 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     1621 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     1681 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     1741 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     1801 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     1861 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     1921 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     1981 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     2041 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     2101 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     2161 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     2221 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     2281 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     2341 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     2401 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     2461 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     2521 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     2581 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     2641 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     2701 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     2761 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     2821 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     2881 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     2941 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     3001 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cac
//
