LOCUS       sequence-192089-        4243 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dsRNA and riboprobe synthesis for Schmidtea mediterranea Hesl-3 
            (hes-like).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4243)
  AUTHORS   Roberts-Galbraith RH, Brubacher JL, Newmark PA
  TITLE     A functional genomics screen in planarians reveals regulators of 
            whole-brain regeneration.
  JOURNAL   Elife. 2016 Sep 9;5. pii: e17002. doi: 10.7554/eLife.17002.
  PUBMED    27612384
REFERENCE   2  (bases 1 to 4243)
  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: "Elife."; 
            date: "2016-09-9"; volume: "5. pii: e17002. doi"; pages: " 
            10.7554/eLife.17002"
COMMENT     Sequence Label: pRRG867-hesl-3
FEATURES             Location/Qualifiers
     source          1..4243
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      14..602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     503..522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     756..773
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(785..832)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        846..864
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        868..886
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     868..885
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(1906..1924)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(1928..1946)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1960..2007
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             2414..3208
                     /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(2468..2487)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3078..3097
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3226..3294,3295..4086)
                     /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(3444..3463)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
       61 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
      121 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
      181 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
      241 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
      301 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
      361 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
      421 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
      481 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
      541 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
      601 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      661 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
      721 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcac
      781 gcgtcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct aggcgcgccg
      841 gatcctaata cgactcacta tagggagatt taggtgacac tatagaagtg accttaaaag
      901 caacaatcca gaactcgatg tagaaaagaa aaaacgccga ggaaaaatcg aaaagcgcag
      961 acgagaccga attaacaaca gcctggcaga gctcaaagaa ttggtaccga atgcaatagc
     1021 aaaacagaat acgacaaaat tggagaaagc tgaaatccta aagctggccg ttgatcactt
     1081 gaaagaatta aagagcaaag attcgagtaa aggttatgga ggatatacaa gcatttatga
     1141 gagcaaaatt gcaggattta gagattgttt aaatgaagtc tcaaagtacc ttcataggga
     1201 aggaatcacg actcaaaatc cactctatca aggcttgatg gagcacccgc agtatttcta
     1261 tcaatgtatc ctttcacaaa ccccttatca gattgaagca ccatcagcga ttgattcggc
     1321 ttttcgatat tcacaaatgc aaactccttt aattacattc gacccgaaca ctgtgccaaa
     1381 ttatcatgaa aactatcaaa agctgacttc gcggagcagc aatgcaattg cgtcttcata
     1441 tcaaaaccag ccgaatcatt atcaagctca aaccagcaat taccaatcaa atttcctttt
     1501 ccaacggaac gcgcatcagc agtatccgac aaacattgcg actccgttga tagcagcgaa
     1561 cgcaaatccg aattccgtat tcaagtgctc atcgactcca tctcggatcg gtttgacaac
     1621 gaatccgtct caaattcaac cgtcaatcca gcagccatct ggactggatt tcccctcgtt
     1681 ttgcaatgat tcgattaatt tcttccagtc cagacaagaa ataccgtaac atgcaaaagt
     1741 gaaaacaaag cctgcaattc actctcctcc atctcctccc ttccgctccc cccttatttt
     1801 atctatccct atctctaccg aattgaatag atttattaaa atatctacag actgtaaata
     1861 catttataaa ctgatttctg tttttcctca ttgatgcaag gtctcccttt agtgagggtt
     1921 aattctccct atagtgagtc gtattaggat ccggcgcgcc tagcataacc ccttggggcc
     1981 tctaaacggg tcttgagggg ttttttgacg cgtcgatttc ggcctattgg ttaaaaaatg
     2041 agctgattta acaaaaattt aacgcgaatt ttaacaaaat tcagggcgca agggctgcta
     2101 aaggaagcgg aacacgtaga aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt
     2161 cagctactgg gctatctgga caagggaaaa cgcaagcgca aagagaaagc aggtagcttg
     2221 cagtgggctt acatggcgat agctagactg ggcggtttta tggacagcaa gcgaaccgga
     2281 attgccagct ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa actggatggc
     2341 tttcttgccg ccaatgatct gatggcgcag gggatcaaga tctgatcaag agacaggatg
     2401 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     2461 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     2521 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     2581 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     2641 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     2701 agtgccgggg caggatctcc tgtcatccca ccttgctcct gccgagaaag tatccatcat
     2761 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     2821 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     2881 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     2941 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     3001 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     3061 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     3121 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     3181 ctatcgcctt cttgacgagt tcttctgaat tgaaaaagga agagtatgag tattcaacat
     3241 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     3301 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     3361 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     3421 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     3481 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     3541 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     3601 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     3661 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     3721 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     3781 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     3841 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     3901 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     3961 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagccag
     4021 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     4081 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     4141 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     4201 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa aga
//
