LOCUS       sequence-486269-        2668 bp    DNA     circular SYN 24-SEP-2025
DEFINITION  Encodes a C-terminal fragment of Spol-foxA to generate dsRNA for 
            RNAi experiments in the planarian S. polychroa.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2668)
  AUTHORS   Deihl EW, Booth CLT, Davies EL
  TITLE     Protocol combining RNA interference and regeneration assays in 
            planarian embryos.
  JOURNAL   bioRxiv [Preprint]. 2025 Jul 22:2025.07.21.666018. doi: 
            10.1101/2025.07.21.666018.
  PUBMED    40777484
REFERENCE   2  (bases 1 to 2668)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 24, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2025 Jul 22:2025.07.21.666018. doi: 
            10.1101/2025.07.21.666018."
COMMENT     Sequence Label: pDL1-Spol-foxA-C-term
FEATURES             Location/Qualifiers
     source          1..2668
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(101..120)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      914..1502
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1403..1422
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1656..1673
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(1680..1727)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1729..1745
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1729..1745
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1746..1764
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1766..1784
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1766..1783
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      1881..1890
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2264..2282)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(2284..2302)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2303..2319)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      2321..2368
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2446..2550
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2551..2619,2620..2668,1..743)
                     /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"
ORIGIN
        1 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
       61 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
      121 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
      181 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
      241 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
      301 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
      361 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
      421 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
      481 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
      541 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
      601 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
      661 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
      721 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
      781 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
      841 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
      901 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
      961 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     1021 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     1081 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     1141 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     1201 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     1261 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     1321 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     1381 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     1441 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     1501 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     1561 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     1621 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgc
     1681 aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctagaca ggaaacagct
     1741 atgactaata cgactcacta tagggattta ggtgacacta tagaaaggca cgttgttgta
     1801 gtttgagtag tatttcatgt gttcgagatc aactgatgag ccagtggctc ctgcgtatag
     1861 agtatcatgg ggactcacac ccatggcggc agcaactagt ctcggatgca tcaaacgatc
     1921 tattgagaac ggattacttg ttgttagtaa tgacgaattg tgttgagatg cttcgtattg
     1981 atatgccata gctgttgata aattgtcata cggatgtccc gataatgata tcgatggtaa
     2041 tgacacagaa atttgatcat cgtttggtaa gttgtgtcct ccataaagtc cgaaaacact
     2101 attcgcacct ggtggcaagt gcataccctg accaagaggg tcatctgatg ccaataagct
     2161 atagtgatct aaagaacttt gttggaaaat gttttgctga tttgagtaga attgctgatt
     2221 gggattgtaa tgaatttgtg gatgttcttg ctggtttggt ctccctttag tgagggttaa
     2281 ttccctatag tgagtcgtat taactggccg tcgttttacc ctagcataac cccttggggc
     2341 ctctaaacgg gtcttgaggg gttttttgcc tgatgcggta ttttctcctt acgcatctgt
     2401 gcggtatttc acaccgcatc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     2461 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2521 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     2581 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     2641 gtaaaagatg ctgaagatca gttgggtg
//
