LOCUS       sequence-460858-        8113 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Negative selection marker for extrachromosomal arrays during RMCE 
            crosses- Psnt-1::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 8113)
  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 8113)
  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: pHW1392
FEATURES             Location/Qualifiers
     source          1..8113
                     /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      5053..5376
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     primer_bind     complement(5394..5410)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5395..5411)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(5435..5454)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5436..5454)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5464..5481)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5464..5480)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5473..5495)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5622..6077
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5709..5728)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5919..5940
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6103..6207
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6208..6276,6277..7068)
                     /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(6426..6445)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7239..7827
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7728..7747
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7981..7998
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    join(8108..8113,1..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 tccttcagaa gacgtgcttt cctttttctt
      241 ctcgtatcga tcccccgtat ggaaggctcc tcgtttcaac agccaacccc ttcccattcg
      301 ttttcgtcgt ccgtcttctt cttcgtcttc ttttttcttt tcttttcctt tttctgctgg
      361 tttttgcgag aaagagtgta tgcgagaaaa gccagttggg gaaaagaaga agaagcggtg
      421 ggaggctcga cggcgcggag cacacttctt ctctgagtct gtctcttgct cacgactcca
      481 gtccaacatc tccagcttct cactaaaacg ttttcttttt tgctcgtgtg tcgtgtgctc
      541 tcaacaaacc gctaaggact ataaaatatt gaataggtgc cagcactttt ttgttctagt
      601 tccagaaaaa gtatttcttg tattggattt tcgattgctt caccgttaca gataattttt
      661 cattgtgaaa aatatcaata tatgggggta ttttgaaaaa tgatattttc atatcgagtt
      721 ttttcaaaat cagtttggtt ttgttggata ttgttcaaaa cgcattttgc ccaaaaaata
      781 aatattgaaa ctttacaaac attatttgta gccccgttcc atgcctattc ctattttccg
      841 ttaaaagcca aaaatacaaa taaaaatgaa aaatctgttt ttcgaacaat ggtattttgg
      901 tctcatcaca cttttgactc ttctcaatta tgcttcgaaa taggaacgaa ttcaaattta
      961 actaatggca cttggggtgc atcttctttt gaaacagaaa actgaccttt cacccaaatg
     1021 aaaattacat gagcttgtga agatgacaca ggtcagtaaa aagaaaaaag tgcgtgggta
     1081 tttctttctc cataattctt gaaatttcat tggagatatt ttgaagaggt gggctcatct
     1141 aaacattaag cttaagattt ttgaaatatt tgaaagaccc aaaataactt acacaatctt
     1201 ttcaacaaaa aaaactgtca tgaattttta tttctgaaaa gtctccaaca ttgtcgacaa
     1261 aggaattgat ttttaatgtc ttcaaataat ttagaagccg acaaatcaaa actgttgatg
     1321 cccttcgctc atctctgatt tatcatcgta tttctcaatt tcccgcgaag ttcgaattta
     1381 gacatgttcc aaggtctccg aaacttttca ccccagaaca tcatacaaat attttataca
     1441 agcttgaaaa aatgttacta agaaatttta tctccagcaa cctaacatat agttactaga
     1501 aacattgttg gctgccaagt tctgagaaac actacaattt ttaaagtttt tccgaaaact
     1561 tttttaaatc tttaagtaac ttatccattc tctcagttct tgaaatatgt tagtgacaag
     1621 ttttaaatat ttttcccaag cctatcttgc aaaactcctg aattagaatc attattcaaa
     1681 gccatgggaa aatcaagaat gcaaccacca ccgcaaatgc tcgatgtgag ctttccccaa
     1741 aatttgcatc ctctacaaca cttctttttc acctgccctc cgaaagagca cctcttctct
     1801 tctttttttt gctctgacga cgacatctaa caaaaaatca attttttttc gtttttccaa
     1861 gcggtccttt cgtgtagtcg acccattcgt attatcattc gatgcttttt cacttcttat
     1921 ttttcgtcgc ttttgaattg catatagcgt gtttatcgca acgaaaaaca gacaaaaaag
     1981 aaagcaaata tgcaaaaaca aaaaaatacg aagccaataa aaaatatcta tcttcaattg
     2041 gaattcgaat tttcaattca aattcaaacc tactaaaacc tactattttc cataattata
     2101 gattctcaaa ggtctccaaa ggaatgagcc aagaacaaaa agttgaggag aatactgcac
     2161 atttcccttt ctataacttt ttcaagatct actaagattt ttgacccttg ggatcctcaa
     2221 agttttcaaa aagtaatcct gctcttctct aaatgacgtt tttttttttg ttttcatctg
     2281 ttcaaaaaac cccttaacta gtcgtcattc ttgcccttga ataataggat aatatgatta
     2341 atctaatgtc ataataatct cctcttcgtc tcccatattt tttcccatac aaggatttca
     2401 tttttacatc cgtgtttctc gtatcagaat gtattcttct tctggaaaag taaacagact
     2461 cctacatctc tcgcctcttt ctgaaaaatt ctaattattt cgcttctgta aacacatttt
     2521 catccggggt ttgcctattt tcccattcga cccccgactc cgcctcttca ttctctcacg
     2581 gaaccctttt cttcgacgct ttgcatcttg atttacctct gatttcattt atcaatcaaa
     2641 ctttcagtca ccatgcaaag cccaactagc aaattggtag aattcaggtg cctgattgcg
     2701 ttggcaatat atttgcacgc gctggagcaa tcgatccagc actgccattg tgttcatggt
     2761 tacagaaata acacggagag cgccgagctg gtctcccact acgagtcgag tctttcgctc
     2821 ccggacattt tgcccattcc ctcaaagacg tacgacaaga accgggctcc caagctcctc
     2881 ggccagccca cagtagtcta cttccatgtc acggtcctct ccctggactc cattaacgag
     2941 gagtctatga cctatgtgac ggacatcttc cttgcacaaa gctggcgtga tcctcgcctg
     3001 cggttgcctg agaacatgag tgagcagtat cgcatattgg atgtcgactg gttgcacagc
     3061 atttggcggc ccgattgctt ctttaagaac gccaaaaagg tcaccttcca tgagatgagc
     3121 attcccaatc actatctctg gctgtaccac gacaaaacgc tgctctatat gtccaaactc
     3181 acgttggtcc tgtcgtgcgc catgaagttt gagtcctatc cgcatgacac gcaaatctgc
     3241 tccatgatga tcgagagttt atcccatacg gtggaagatt tggttttcat ttggaacatg
     3301 accgacccac ttgtggttaa cacggagatt gagttgccgc agctagacat atcaaataac
     3361 tacacaaccg actgtactat agagtactca acaggtaact tcacctgcct ggccattgtg
     3421 ttcaacctgc gccgacgcct gggttaccat ttgttccaca cctacatccc ctcggctctg
     3481 attgtggtca tgtcttggat atcgttttgg ataaaaccag aagcgatacc cgcccgtgta
     3541 actctgggag tgacctcact gctaaccctg gccacccaga atacccagtc gcaacaatcg
     3601 ctgccgccgg tttcgtatgt caaggctata gacgtctgga tgtcgtcctg ttcggtgttt
     3661 gtattccttt ctctgatgga atttgcagtg gtcaacaatt ttatgggacc ggtggccaca
     3721 aaggcaatga aggggtattc ggacgagaac atcagtgatc tggacgacct aaagcatcat
     3781 cgggaatcga ttattgagcc ccagtacgac actttctgcc atggccatgc cacagccatt
     3841 tatatagaca aattctcgcg ctttttcttc ccgttttcgt tctttatact caatattgtc
     3901 tattggacaa cgttcctaaa gtaagctgtc tcatcctact ttcacctagt taactgcttg
     3961 tcttaaaatc tatgcttctc tttagtatct aaaattttcc tagaagctta caagtatata
     4021 aatggtctct tctcaataaa ggttgtatat ttattcatct tattgaatct gccatttcct
     4081 cgtttttgcg agtttatata ccttccaatt ttctttctat tgtattttca acttctaatt
     4141 ttaattcagg gaaactgctt caacgcatca aaaatgggta tggtcagtaa aggagaagaa
     4201 ttgttcactg gagttgtccc aatcctcgtc gagctcgacg gagacgtcaa cggacacaag
     4261 ttctccgtct ccggagaggg agagggagac gccacctacg gaaagctcac cctcaagttc
     4321 atctgcacca ccggaaagct cccagtccca tggccaaccc tcgtcaccac cttctgctac
     4381 ggagtccaat gcttctcccg ttacccagac cacatgaagc gtcacgactt cttcaagtcc
     4441 gccatgccag agggatacgt ccaagagcgt accatcttct tcaaggtaag tttaaacata
     4501 tatatactaa ctactgatta tttaaatttt caggacgacg gaaactacaa gacccgtgcc
     4561 gaggtcaagt tcgagggaga caccctcgtc aaccgtatcg agctcaaggt aagtttaaac
     4621 agttcggtac taactaacca tacatattta aattttcagg gaatcgactt caaggaggac
     4681 ggaaacatcc tcggacacaa gctcgagtac aactacaact cccacaacgt ctacatcatg
     4741 gccgacaagc aaaagaacgg aatcaaggtc aacttcaagg taagtttaaa catgatttta
     4801 ctaactaact aatctgattt aaattttcag atccgtcaca acatcgagga cggatccgtc
     4861 caactcgccg accactacca acaaaacacc ccaatcggag acggaccagt cctcctccca
     4921 gacaaccact acctctccac ccaatccgcc ctctccaagg acccaaacga gaagcgtgac
     4981 cacatggtcc tcctcgagtt cgtcaccgcc gccggaatca cccacggaat ggacgagctc
     5041 tacaagacgt aaatgcaaaa tcctttcaag cattcccttc ttctctatca ctcttctttc
     5101 tttttgtcaa aaaattctct cgctaattta tttgcttttt taatgttatt attttatgac
     5161 tttttatagt cactgaaaag tttgcatctg agtgaagtga atgctatcaa aatgtgattc
     5221 tgtctgatgt actttcacaa tctctcttca attccatttt gaagtgcttt aaacccgaaa
     5281 ggttgagaaa aatgcgagcg ctcaaatatt tgtattgtgt tcgttgagtg acccaacaaa
     5341 aagaggaaac tttattgtgc cgccaagaaa aaagtctcag taatcaagct tatcgatacc
     5401 gtcgacctcg agggggggcc cggtacccaa ttcgccctat agtgagtcgt attacgcgcg
     5461 ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     5521 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     5581 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
     5641 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     5701 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     5761 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     5821 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     5881 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     5941 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     6001 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     6061 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     6121 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     6181 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     6241 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     6301 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     6361 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     6421 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     6481 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     6541 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6601 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6661 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6721 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     6781 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     6841 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6901 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6961 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     7021 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     7081 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     7141 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     7201 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     7261 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     7321 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     7381 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     7441 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     7501 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     7561 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7621 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7681 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7741 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7801 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7861 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     7921 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     7981 agcgagtcag tgagcgagga agcggcccaa tacgcaaacc gcctctcccc gcgcgttggc
     8041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     8101 acgcaattaa tgt
//
