LOCUS       sequence-460828-        3432 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination RMCE vector containing FRT::SA::51-bp Artificial 
            exon::3xStop::SL2::SapI Insertion sites (to clone new driver with 
            3'UTR in slot 5 and SEC in slot 6)::reverse FRT3 to swap driver via 
            RMCE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3432)
  AUTHORS   Khan H, Huang X, Raj V, Wang H
  TITLE     A versatile site-directed gene trap strategy to manipulate gene 
            activity and control gene expression in Caenorhabditis elegans.
  JOURNAL   PLoS Genet. 2025 Jan 22;21(1):e1011541. doi: 
            10.1371/journal.pgen.1011541. eCollection 2025 Jan.
  PUBMED    39841730
REFERENCE   2  (bases 1 to 3432)
  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: "PLoS 
            Genet."; date: "2025-01-22"; volume: "21(1):e1011541. doi"; pages: "
            10.1371/journal.pgen.1011541. eCollection 2025 Jan"
COMMENT     Sequence Label: pHW1133
FEATURES             Location/Qualifiers
     source          1..3432
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    38..54
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     43..65
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     62..78
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     62..78
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     97..117
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        99..117
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    185..218
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             259..282
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    complement(631..664)
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     primer_bind     complement(682..698)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(683..699)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(723..742)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(724..742)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(752..769)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(752..768)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(761..783)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      910..1365
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(997..1016)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1207..1228
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1391..1495
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1496..1564,1565..2356)
                     /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(1714..1733)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2527..3115
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3016..3035
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3269..3286
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3396..3417
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3432,1..5,6..23,24..30)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
       61 acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa
      121 caaaagctgg agctccaccg cggtggcggc cgctctagaa gggctgcagg aattcgatgc
      181 tagcgaagtt cctattctct agaaagtata ggaacttcta actaatctga tttaaatttt
      241 cagggtaccg gcggaagcga ctacaaggac gacgacgaca aggagccaac cacttgacta
      301 agtgataagc tagctaagct gtctcatcct actttcacct agttaactgc ttgtcttaaa
      361 atctatgctt ctctttagta tctaaaattt tcctagaagc ttacaagtat ataaatggtc
      421 tcttctcaat aaaggttgta tatttattca tcttattgaa tctgccattt cctcgttttt
      481 gcgagtttat ataccttcca attttctttc tattgtattt tcaacttcta attttaattc
      541 agggaaactg cttcaacgca tcaaaaatgc gaagagccca tggatccact agtcatatgg
      601 aattctgcag gcctgctctt cgggtcccgg gaagttccta tactatttga agaataggaa
      661 cttccccggg atcaagctta tcgataccgt cgacctcgag ggggggcccg gtacccaatt
      721 cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact
      781 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
      841 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
      901 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
      961 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     1021 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     1081 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     1141 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     1201 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     1261 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     1321 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     1381 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     1441 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     1501 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     1561 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     1621 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     1681 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     1741 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     1801 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     1861 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     1921 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     1981 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     2041 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     2101 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     2161 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     2221 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     2281 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     2341 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     2401 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     2461 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2521 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2581 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2641 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     2701 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2761 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2821 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2881 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     2941 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3001 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3061 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3121 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     3181 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     3241 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggcccaata
     3301 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     3361 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     3421 gcaccccagg ct
//
