LOCUS       sequence-460857-        8095 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Negative selection marker for extrachromosomal arrays during RMCE 
            crosses - Psnt-1::HisCl1 cDNA::SL2::mScarlet-I-C1::tbb-2 3'UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8095)
  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 8095)
  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: pHW1391
FEATURES             Location/Qualifiers
     source          1..8095
                     /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      5035..5358
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     primer_bind     complement(5376..5392)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5377..5393)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(5417..5436)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5418..5436)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5446..5463)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5446..5462)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5455..5477)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5604..6059
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5691..5710)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5901..5922
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6085..6189
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6190..6258,6259..7050)
                     /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(6408..6427)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7221..7809
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7710..7729
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7963..7980
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    join(8090..8095,1..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 tggtttccaa gggagaggcc
     4201 gtcatcaagg agttcatgcg tttcaaggtc cacatggagg gatccatgaa cggacacgag
     4261 ttcgagatcg agggagaggg agagggacgt ccatacgagg gaacccaaac cgccaagctc
     4321 aaggtcacca aggtaagttt aaacatatat atactaacta accctgatta tttaaatttt
     4381 cagggaggac cactcccatt ctcctgggac atcctctccc cacaattcat gtacggatcc
     4441 cgtgccttca tcaagcaccc agccgacatc ccagactact acaagcaatc cttcccagag
     4501 ggattcaagt gggagcgtgt catgaacttc gaggacggag gagccgtcac cgtcacccaa
     4561 gacacctccc tcgaggacgg aaccctcatc tacaaggtaa gtttaaacag ttcggtacta
     4621 actaaccata catatttaaa ttttcaggtc aagctccgtg gaaccaactt cccaccagac
     4681 ggaccagtca tgcaaaagaa gaccatggga tgggaggcct ccaccgagcg tctctaccca
     4741 gaggacggag tcctcaaggg agacatcaag atggccctcc gtctcaagga cggaggacgt
     4801 tacctcgccg acttcaaggt aagtttaaac atgattttac taactaacta atctgattta
     4861 aattttcaga ccacctacaa ggccaagaag ccagtccaaa tgccaggagc ctacaacgtc
     4921 gaccgtaagc tcgacatcac ctcccacaac gaggactaca ccgtcgtcga gcaatacgag
     4981 cgttccgagg gacgtcactc caccggagga atggacgagc tctacaagac gtaaatgcaa
     5041 aatcctttca agcattccct tcttctctat cactcttctt tctttttgtc aaaaaattct
     5101 ctcgctaatt tatttgcttt tttaatgtta ttattttatg actttttata gtcactgaaa
     5161 agtttgcatc tgagtgaagt gaatgctatc aaaatgtgat tctgtctgat gtactttcac
     5221 aatctctctt caattccatt ttgaagtgct ttaaacccga aaggttgaga aaaatgcgag
     5281 cgctcaaata tttgtattgt gttcgttgag tgacccaaca aaaagaggaa actttattgt
     5341 gccgccaaga aaaaagtctc agtaatcaag cttatcgata ccgtcgacct cgaggggggg
     5401 cccggtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt
     5461 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     5521 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     5581 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     5641 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     5701 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     5761 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     5821 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     5881 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     5941 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     6001 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     6061 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     6121 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     6181 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     6241 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     6301 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     6361 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     6421 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     6481 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     6541 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     6601 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6661 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6721 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     6781 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     6841 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6901 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     6961 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     7021 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     7081 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     7141 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     7201 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     7261 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     7321 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     7381 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7441 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7501 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7561 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7621 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7681 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7741 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7801 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7861 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     7921 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     7981 gaagcggccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     8041 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgt
//
