LOCUS       sequence-193554-        3759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dsRNA and riboprobe synthesis for Schmidtea mediterranea sFRP1 
            (secreted frizzled related protein 1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3759)
  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 3759)
  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: pRRG43-sFRP1
FEATURES             Location/Qualifiers
     source          1..3759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      13..601
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     502..521
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     755..772
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(784..831)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        845..863
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        867..885
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     867..884
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(1426..1444)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(1448..1466)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1480..1527
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             1929..2723
                     /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(1983..2002)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2593..2612
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(2741..2809,2810..3601)
                     /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(2959..2978)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
       61 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
      121 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
      181 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
      241 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
      301 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
      361 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
      421 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
      481 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
      541 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
      601 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
      661 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
      721 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcacg
      781 cgtcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta ggcgcgccgg
      841 atcctaatac gactcactat agggagattt aggtgacact atagaagtga ccttaaacac
      901 tcaatgccat gtcgacataa aaaagttcat ttgttcactt tatgctccgg tatgcataag
      961 caatcagaga aatcaaaagg ttccgccttg cagagaactc tgcgtgactg ttaaggaatc
     1021 gtgtttgccg agtatgaaaa tgtacggatt tgactggcca acaattatga aatgttcgaa
     1081 atttccacaa tttaaaaatt cattatgtat accaaaaacc gtcgtgccag gtaaaaagtg
     1141 tgacctatgc atgaaaatag cttcctatga agtcattgct aacagattct gtttatctcc
     1201 catagtaatc agagcgaaaa tcaagagaat tattccaacc agtggcaatg ccattcagat
     1261 tattctacaa aagaaatcga aattcctcaa atttcaagcc aatttcagtc agaacgctct
     1321 ggagttgaat ctgaaatgca actgtacgaa tctcaaattc aaacaacgcg ccttgaaagg
     1381 ccggtggatt attatggcac aaatcgacag cgacaacaag gtctcccttt agtgagggtt
     1441 aattctccct atagtgagtc gtattaggat ccggcgcgcc tagcataacc ccttggggcc
     1501 tctaaacggg tcttgagggg ttttttgtcg atttcggcct attggttaaa aaatgagctg
     1561 atttaacaaa aatttaacgc gaattttaac aaaattcagg gcgcaagggc tgctaaagga
     1621 agcggaacac gtagaaagcc agtccgcaga aacggtgctg accccggatg aatgtcagct
     1681 actgggctat ctggacaagg gaaaacgcaa gcgcaaagag aaagcaggta gcttgcagtg
     1741 ggcttacatg gcgatagcta gactgggcgg ttttatggac agcaagcgaa ccggaattgc
     1801 cagctggggc gccctctggt aaggttggga agccctgcaa agtaaactgg atggctttct
     1861 tgccgccaat gatctgatgg cgcaggggat caagatctga tcaagagaca ggatgaggat
     1921 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     1981 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     2041 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     2101 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     2161 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     2221 cggggcagga tctcctgtca tcccaccttg ctcctgccga gaaagtatcc atcatggctg
     2281 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     2341 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     2401 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     2461 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     2521 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     2581 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     2641 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     2701 gccttcttga cgagttcttc tgaattgaaa aaggaagagt atgagtattc aacatttccg
     2761 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     2821 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     2881 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     2941 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     3001 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     3061 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     3121 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     3181 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     3241 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     3301 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     3361 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     3421 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     3481 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gccaggcaac
     3541 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     3601 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     3661 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     3721 gttttcgttc cactgagcgt cagaccccgt agaaaagat
//
