LOCUS       sequence-193552-        3656 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dsRNA and riboprobe synthesis for Schmidtea mediterranea ChAT 
            (choline acetyltransferase).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3656)
  AUTHORS   Roberts-Galbraith RH, Newmark PA
  TITLE     Follistatin antagonizes activin signaling and acts with notum to 
            direct planarian head regeneration.
  JOURNAL   Proc Natl Acad Sci U S A. 2013 Jan 22;110(4):1363-8. doi: 
            10.1073/pnas.1214053110. Epub 2013 Jan 7.
  PUBMED    23297191
REFERENCE   2  (bases 1 to 3656)
  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"; doi: 
            "10.1073/pnas.1214053110"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2013-01-22- 22"; volume: "110"; issue: "4"; pages: "1363-8"
COMMENT     Sequence Label: pRRG36-ChAT
FEATURES             Location/Qualifiers
     source          1..3656
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      5..593
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     494..513
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(759..806)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        820..838
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        842..860
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     842..859
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     RBS             1120..1128
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(1310..1328)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(1332..1350)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1364..1411
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             1818..2612
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1872..1891)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2482..2501
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(2630..2698,2699..3490)
                     /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     complement(2848..2867)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
       61 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
      121 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
      181 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
      241 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
      301 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
      361 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
      421 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
      481 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
      541 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
      601 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
      661 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
      721 ctcgccgcag ccgaacgacc gagcgcagcg agacgcgtca aaaaacccct caagacccgt
      781 ttagaggccc caaggggtta tgctaggcgc gccggatcct aatacgactc actataggga
      841 gatttaggtg acactataga agtgaccttg atctcctgcc accaaagaaa ctagaatata
      901 atattgatag ctttgccagt caaatgatat ctgaagccaa acaagaaatt aaaaatttgg
      961 ccgataattt ggatatttat attctacgat accagaaatt tggtagagat tttcctaaaa
     1021 ctgtcaacat gagtcctgat tccttcatcc aactggcact tcaactcacc cattacaaga
     1081 ttcataaatg tcttgttcca acatatgaaa gtgcttccat aaggaggttt tctctgggac
     1141 gcgtagacaa cattcgtgcg gcttcaattc cagcattaaa atggtgtaca tcaatgactg
     1201 ctgatactcc tgcttcggat gacgaaaaga tcattctttt acgctcggca atggaatggc
     1261 agacaaaggt tatgttggac acgattcagg gtcatggagt taaggtctcc ctttagtgag
     1321 ggttaattct ccctatagtg agtcgtatta ggatccggcg cgcctagcat aaccccttgg
     1381 ggcctctaaa cgggtcttga ggggtttttt gacgcgtcga tttcggccta ttggttaaaa
     1441 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaattcaggg cgcaagggct
     1501 gctaaaggaa gcggaacacg tagaaagcca gtccgcagaa acggtgctga ccccggatga
     1561 atgtcagcta ctgggctatc tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag
     1621 cttgcagtgg gcttacatgg cgatagctag actgggcggt tttatggaca gcaagcgaac
     1681 cggaattgcc agctggggcg ccctctggta aggttgggaa gccctgcaaa gtaaactgga
     1741 tggctttctt gccgccaatg atctgatggc gcaggggatc aagatctgat caagagacag
     1801 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     1861 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     1921 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     1981 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     2041 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     2101 gcgaagtgcc ggggcaggat ctcctgtcat cccaccttgc tcctgccgag aaagtatcca
     2161 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     2221 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     2281 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     2341 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     2401 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     2461 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     2521 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     2581 ccttctatcg ccttcttgac gagttcttct gaattgaaaa aggaagagta tgagtattca
     2641 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     2701 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     2761 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     2821 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     2881 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     2941 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     3001 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     3061 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     3121 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     3181 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     3241 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     3301 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     3361 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     3421 ccaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     3481 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     3541 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     3601 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggat
//
