LOCUS       sequence-460861-        6649 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Negative selection marker for extrachromosomal arrays during RMCE 
            crosses - Pmyo-2::HisCl1 cDNA::SL2::GFP-C1::tbb-2 3'UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6649)
  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 6649)
  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: pHW1387
FEATURES             Location/Qualifiers
     source          1..6649
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(31..36,37..54,55..61)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    69..85
                     /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     74..96
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     93..109
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     93..109
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     128..148
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        130..148
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     terminator      3589..3912
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     primer_bind     complement(3930..3946)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3931..3947)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(3971..3990)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3972..3990)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4000..4017)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4000..4016)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4009..4031)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4158..4613
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4245..4264)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4455..4476
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4639..4743
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4744..4812,4813..5604)
                     /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(4962..4981)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5775..6363
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6264..6283
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6517..6534
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    join(6644..6649,1..16)
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
       61 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
      121 caagcgcgca attaaccctc actaaaggga acaaaagctg gagctccacc gcggtggcgg
      181 ccgctctaga agggctgcag gaattcgatt ggcattttat atctgagtag tatcctttgc
      241 tttaaatgtc cataaaacta attttataat caataaaaca acgtttgtaa atcaactgag
      301 tttacaagta gagacattga gggatacttt cactatgcta aagtgataat cgaccaaata
      361 ataacttcac tttggtattt attcctgtct tataaatgtt atgtatgaat taaattcata
      421 tgcatatggc tcactctgac aatttttaat aatcttccag atcaatattg actaccgatg
      481 cgggtggtct tttgctttga attctgctga actttacacc ccgaacagca atgtgtgctt
      541 cagcctaaaa aaaagtaagt gtgttaatca gtgcccccga ttcttcattt tttgcccctc
      601 tctcccgttt cgtcggcaaa agaagagaaa ataaagataa gtctcaagat aggttggtaa
      661 tcgctaaagt ggttgtgtgg ataagagtag caaaatggca ggaggagcac tttgcgcgca
      721 cacactgtac tcattgttct ggataaaatt ctctcgttgt ttgccgtcgg atgtctgcct
      781 ctctgcattg agccggcttc ttcactatct ttagttaacc taaaatgccg tttcttttct
      841 cgtatcccca ctatcccgtt gaggttctct gctctcttcg ctccctaccg ccagcgagca
      901 actatccgtg ggggcgcctt gctcggaaga tgggggggaa gaaagaagat ttttgctatt
      961 tgcacttgag aaagagactt ttcctgcgtc gatggtcaga gaacagtgtg cagacacttt
     1021 tcagctacct agaattacaa ttggatatcc ccgcctccca atccacccac ccagggaaaa
     1081 agaagggctc gccgaaaatc aaagttatct ccaggctcgc gcatcccacc gagcggttga
     1141 cttctctcca ccacttttca ttttaaccct cgatcgtcag acacagaaat gcaaagccca
     1201 actagcaaat tggtagaatt caggtgcctg attgcgttgg caatatattt gcacgcgctg
     1261 gagcaatcga tccagcactg ccattgtgtt catggttaca gaaataacac ggagagcgcc
     1321 gagctggtct cccactacga gtcgagtctt tcgctcccgg acattttgcc cattccctca
     1381 aagacgtacg acaagaaccg ggctcccaag ctcctcggcc agcccacagt agtctacttc
     1441 catgtcacgg tcctctccct ggactccatt aacgaggagt ctatgaccta tgtgacggac
     1501 atcttccttg cacaaagctg gcgtgatcct cgcctgcggt tgcctgagaa catgagtgag
     1561 cagtatcgca tattggatgt cgactggttg cacagcattt ggcggcccga ttgcttcttt
     1621 aagaacgcca aaaaggtcac cttccatgag atgagcattc ccaatcacta tctctggctg
     1681 taccacgaca aaacgctgct ctatatgtcc aaactcacgt tggtcctgtc gtgcgccatg
     1741 aagtttgagt cctatccgca tgacacgcaa atctgctcca tgatgatcga gagtttatcc
     1801 catacggtgg aagatttggt tttcatttgg aacatgaccg acccacttgt ggttaacacg
     1861 gagattgagt tgccgcagct agacatatca aataactaca caaccgactg tactatagag
     1921 tactcaacag gtaacttcac ctgcctggcc attgtgttca acctgcgccg acgcctgggt
     1981 taccatttgt tccacaccta catcccctcg gctctgattg tggtcatgtc ttggatatcg
     2041 ttttggataa aaccagaagc gatacccgcc cgtgtaactc tgggagtgac ctcactgcta
     2101 accctggcca cccagaatac ccagtcgcaa caatcgctgc cgccggtttc gtatgtcaag
     2161 gctatagacg tctggatgtc gtcctgttcg gtgtttgtat tcctttctct gatggaattt
     2221 gcagtggtca acaattttat gggaccggtg gccacaaagg caatgaaggg gtattcggac
     2281 gagaacatca gtgatctgga cgacctaaag catcatcggg aatcgattat tgagccccag
     2341 tacgacactt tctgccatgg ccatgccaca gccatttata tagacaaatt ctcgcgcttt
     2401 ttcttcccgt tttcgttctt tatactcaat attgtctatt ggacaacgtt cctaaagtaa
     2461 gctgtctcat cctactttca cctagttaac tgcttgtctt aaaatctatg cttctcttta
     2521 gtatctaaaa ttttcctaga agcttacaag tatataaatg gtctcttctc aataaaggtt
     2581 gtatatttat tcatcttatt gaatctgcca tttcctcgtt tttgcgagtt tatatacctt
     2641 ccaattttct ttctattgta ttttcaactt ctaattttaa ttcagggaaa ctgcttcaac
     2701 gcatcaaaaa tgggtatggt cagtaaagga gaagaattgt tcactggagt tgtcccaatc
     2761 ctcgtcgagc tcgacggaga cgtcaacgga cacaagttct ccgtctccgg agagggagag
     2821 ggagacgcca cctacggaaa gctcaccctc aagttcatct gcaccaccgg aaagctccca
     2881 gtcccatggc caaccctcgt caccaccttc tgctacggag tccaatgctt ctcccgttac
     2941 ccagaccaca tgaagcgtca cgacttcttc aagtccgcca tgccagaggg atacgtccaa
     3001 gagcgtacca tcttcttcaa ggtaagttta aacatatata tactaactac tgattattta
     3061 aattttcagg acgacggaaa ctacaagacc cgtgccgagg tcaagttcga gggagacacc
     3121 ctcgtcaacc gtatcgagct caaggtaagt ttaaacagtt cggtactaac taaccataca
     3181 tatttaaatt ttcagggaat cgacttcaag gaggacggaa acatcctcgg acacaagctc
     3241 gagtacaact acaactccca caacgtctac atcatggccg acaagcaaaa gaacggaatc
     3301 aaggtcaact tcaaggtaag tttaaacatg attttactaa ctaactaatc tgatttaaat
     3361 tttcagatcc gtcacaacat cgaggacgga tccgtccaac tcgccgacca ctaccaacaa
     3421 aacaccccaa tcggagacgg accagtcctc ctcccagaca accactacct ctccacccaa
     3481 tccgccctct ccaaggaccc aaacgagaag cgtgaccaca tggtcctcct cgagttcgtc
     3541 accgccgccg gaatcaccca cggaatggac gagctctaca agacgtaaat gcaaaatcct
     3601 ttcaagcatt cccttcttct ctatcactct tctttctttt tgtcaaaaaa ttctctcgct
     3661 aatttatttg cttttttaat gttattattt tatgactttt tatagtcact gaaaagtttg
     3721 catctgagtg aagtgaatgc tatcaaaatg tgattctgtc tgatgtactt tcacaatctc
     3781 tcttcaattc cattttgaag tgctttaaac ccgaaaggtt gagaaaaatg cgagcgctca
     3841 aatatttgta ttgtgttcgt tgagtgaccc aacaaaaaga ggaaacttta ttgtgccgcc
     3901 aagaaaaaag tctcagtaat caagcttatc gataccgtcg acctcgaggg ggggcccggt
     3961 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
     4021 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     4081 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     4141 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4201 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4261 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4321 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4381 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4441 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4501 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4561 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     4621 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4681 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4741 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4801 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4861 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4921 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4981 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5041 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5101 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5161 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5221 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5281 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5341 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5401 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5461 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5521 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5581 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     5641 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5701 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5761 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5821 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5881 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5941 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6001 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6061 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6121 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6181 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6241 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6301 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6361 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6421 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     6481 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     6541 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     6601 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgt
//
