LOCUS       sequence-426489-        4407 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty repair plasmid for ChrIII split hygromycinR landing pad.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4407)
  AUTHORS   Moerdyk-Schauwecker MJ, Jahahn EK, Munoz ZI, Robinson KJ, Phillips 
            PC
  TITLE     Expansion of the split hygromycin toolkit for transgene insertion in
            Caenorhabditis elegans.
  JOURNAL   MicroPubl Biol. 2024 Jan 29;2024:10.17912/micropub.biology.001091. 
            doi: 10.17912/micropub.biology.001091. eCollection 2024.
  PUBMED    38351905
REFERENCE   2  (bases 1 to 4407)
  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: "MicroPubl 
            Biol."; date: "2024-01-29"; volume: 
            "2024:10.17912/micropub.biology.001091. doi"; pages: " 
            10.17912/micropub.biology.001091. eCollection 2024"
COMMENT     Sequence Label: pMS158
FEATURES             Location/Qualifiers
     source          1..4407
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      340..928
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     829..848
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1082..1099
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1216..1237
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1252..1257,1258..1275,1276..1282)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1290..1306
                     /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     1295..1317
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1314..1330
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1314..1330
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    complement(2208..2241)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     primer_bind     complement(3346..3365)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3465..3487
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3525..3543)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3611..3715
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3716..3784,3785..4407,1..169)
                     /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(3934..3953)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
       61 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
      121 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
      181 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
      241 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
      301 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
      361 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
      421 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
      481 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
      541 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtagtg
      601 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
      661 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
      721 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
      781 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
      841 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
      901 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
      961 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     1021 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     1081 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     1141 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     1201 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     1261 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     1321 cagctatgac catgattacg ccaagcttgt aaatggtcgg taaagatcaa tatcaaattg
     1381 ggctctgagg gaatgactga aaatccactt tttctgcttg aactcagcag atataatttt
     1441 tttaaaaatt tagtttttaa attctgaaag aacacaaaat ttgcaacaaa atggctacag
     1501 ggatttttga aaaagttagt ttgaagccag atattaattt ttaaaaattg gaaaaactac
     1561 taattttgca tataaatggt cggtttaatc aatttcaaat tgtgcactga aatgtatgat
     1621 taaatatcca ctttttctgc ttgaacttag cggatatgaa acttttttaa atttagtttt
     1681 taaattctga aagaacacaa aatttgcaac aaaatagcta cagggatttt tgaaaaagtt
     1741 agtttgaagc cagatattca tttttaaagt ttggaaaatc tacatcgtta actaatttct
     1801 gaaaaacagg aaaaagtcaa ataaatcgca aaaatggaag ttcaaataaa aaagttttga
     1861 aatttttcct aatttttcaa aaacttatgc aatgtgtatt aaatgtataa atgtataaat
     1921 tatgtattaa tatgttgtgt aataaaaatg tattacctat caattttgaa gcaaagtagt
     1981 ttgaaatttt aatttcagga aatttttgca ccaacttact tcaaacttaa tcgatttttt
     2041 tttcgttttt tctttaaaaa tcattttcaa tggttttacc tgaatctata cagttaatgc
     2101 atttgtatgt tcgttaaaat tcattgtaaa tgaccttccc atatgaagga ccgtgtattc
     2161 gaataatgcc attagtattc tcaacgaatt ccttaagtaa gtttaaaata acttcgtata
     2221 ggatacttta tacgaagtta tatttttgct ttcgtcgtaa atctacacac gcgtctcttc
     2281 cgtgcgagag tccaagccag cagccaaatt cgttgactga gtattcaacg tttatacgtt
     2341 gtcggcaacg agaaatagga aaatgcatcg ggaaatgttc ttttttcgat tttttccaag
     2401 gttttgacaa attttaccac gaattttgct atgttttcaa ttaaaaaata tgttattcaa
     2461 ctgtttctat gaggaaaata aggctttgca tgtaattttc ttattcagca taatttttaa
     2521 ttaatttgaa ttttctgtcc taacgtttat tttgttttct tggttatgac tgatctgaaa
     2581 ttaatttttg aattttaagg taatatgaaa aaacccgagt tgaccgccac atccgtagag
     2641 aagttcctca tcgagaagtt cgactccgtc tccgacctca tgcaactctc cgagggagag
     2701 gagtcccgtg ccttctcctt cgacgtcgga ggacgtggat acgtcctccg tgtcaactcc
     2761 tgcgccgacg gattctacaa ggaccgttac gtctaccgtc acttcgcctc cgccgccctc
     2821 ccaatcccag aggtcctcga catcggagag ttctccgagt ccctcaccta ctgcatctcc
     2881 cgtcgtgccc aaggagtcac cctccaagac ctcccagaga ccgagctccc agccgtcctc
     2941 caaccagtcg ccgaggccat ggacgccatc gccgccgccg acctctccca aacctccgga
     3001 ttcggaccat tcggaccaca aggaatcgga caatacacca cctggcgtga cttcatctgc
     3061 gccatcgccg acccacacgt ctaccactgg caaaccgtca tggacgacac cgtctccgcc
     3121 tccgtcgccc aagccctcga cgagctcatg ctctgggccg aggactgccc agaggtccgt
     3181 cacctcgtcc acgccgactt cggatccaac aacgtcctca ccgacaacgg acgtatcacc
     3241 gccgtcatcg actggtccga ggccatgttc ggagactccc aatatgcggt attttctcct
     3301 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
     3361 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     3421 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     3481 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     3541 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     3601 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3661 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     3721 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3781 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     3841 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3901 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     3961 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4021 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4081 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4141 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4201 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4261 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4321 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4381 ccttccggct ggctggttta ttgctga
//
