LOCUS       sequence-460859-        6631 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Negative selection marker for extrachromosomal arrays during RMCE 
            crosses - Pmyo-2::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 6631)
  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 6631)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pHW1385
FEATURES             Location/Qualifiers
     source          1..6631
                     /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      3571..3894
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     primer_bind     complement(3912..3928)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3913..3929)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(3953..3972)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3954..3972)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3982..3999)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3982..3998)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3991..4013)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4140..4595
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4227..4246)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4437..4458
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4621..4725
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4726..4794,4795..5586)
                     /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(4944..4963)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5757..6345
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6246..6265
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6499..6516
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    join(6626..6631,1..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 ttccaaggga gaggccgtca tcaaggagtt catgcgtttc
     2761 aaggtccaca tggagggatc catgaacgga cacgagttcg agatcgaggg agagggagag
     2821 ggacgtccat acgagggaac ccaaaccgcc aagctcaagg tcaccaaggt aagtttaaac
     2881 atatatatac taactaaccc tgattattta aattttcagg gaggaccact cccattctcc
     2941 tgggacatcc tctccccaca attcatgtac ggatcccgtg ccttcatcaa gcacccagcc
     3001 gacatcccag actactacaa gcaatccttc ccagagggat tcaagtggga gcgtgtcatg
     3061 aacttcgagg acggaggagc cgtcaccgtc acccaagaca cctccctcga ggacggaacc
     3121 ctcatctaca aggtaagttt aaacagttcg gtactaacta accatacata tttaaatttt
     3181 caggtcaagc tccgtggaac caacttccca ccagacggac cagtcatgca aaagaagacc
     3241 atgggatggg aggcctccac cgagcgtctc tacccagagg acggagtcct caagggagac
     3301 atcaagatgg ccctccgtct caaggacgga ggacgttacc tcgccgactt caaggtaagt
     3361 ttaaacatga ttttactaac taactaatct gatttaaatt ttcagaccac ctacaaggcc
     3421 aagaagccag tccaaatgcc aggagcctac aacgtcgacc gtaagctcga catcacctcc
     3481 cacaacgagg actacaccgt cgtcgagcaa tacgagcgtt ccgagggacg tcactccacc
     3541 ggaggaatgg acgagctcta caagacgtaa atgcaaaatc ctttcaagca ttcccttctt
     3601 ctctatcact cttctttctt tttgtcaaaa aattctctcg ctaatttatt tgctttttta
     3661 atgttattat tttatgactt tttatagtca ctgaaaagtt tgcatctgag tgaagtgaat
     3721 gctatcaaaa tgtgattctg tctgatgtac tttcacaatc tctcttcaat tccattttga
     3781 agtgctttaa acccgaaagg ttgagaaaaa tgcgagcgct caaatatttg tattgtgttc
     3841 gttgagtgac ccaacaaaaa gaggaaactt tattgtgccg ccaagaaaaa agtctcagta
     3901 atcaagctta tcgataccgt cgacctcgag ggggggcccg gtacccaatt cgccctatag
     3961 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     4021 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     4081 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     4141 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4201 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4261 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4321 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4381 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4441 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4501 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4561 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     4621 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4681 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     4741 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     4801 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     4861 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     4921 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     4981 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5041 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5101 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5161 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5221 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     5281 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5341 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5401 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5461 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5521 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5581 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     5641 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5701 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5761 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5821 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5881 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     5941 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6001 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6061 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6121 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6181 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6241 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6301 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6361 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6421 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     6481 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggcccaata cgcaaaccgc
     6541 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     6601 aagcgggcag tgagcgcaac gcaattaatg t
//
