LOCUS       sequence-426501-        4194 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty repair plasmid for ChrI split hygromycinR landing pad.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4194)
  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 4194)
  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: pMS114
FEATURES             Location/Qualifiers
     source          1..4194
                     /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(1974..2007)
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AGGTATAC) (Shaw et al., 2021). loxN sites are 
                     compatible with each other, but incompatible with loxP or 
                     lox2272 sites."
     primer_bind     complement(3133..3152)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3252..3274
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3312..3330)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3398..3502
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3503..3571,3572..4194,1..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(3721..3740)
                     /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 ccaagctgta acgtacatat ggtgccaaaa tggccaaata
     1381 tcatagtaaa atttttcaaa acatttttga aaaaagtggc aattactgag tttttgccac
     1441 ttttttttag taaactatag ctctctcgca tactcgaatt cctacaaaaa tttatatttg
     1501 aaatataagt accataaaat ccggagaaaa acttccaaaa aaatggcaaa aactaacatt
     1561 cttccaattt tcaaaaaatc atataaaatt tggaattttt taaagaattt ttatactatg
     1621 atatttggac ctttaagctg cgtttttgta gttgacaatt taaaaaaaaa ctccaaattg
     1681 ctttgaaaac taccaaaatg agtaccctga taaccatatt gaaaaaccat caaatgggta
     1741 atgtctagaa aaatgaacga aaaattagag aaaataggaa atgagatatg ttcttcttcc
     1801 tattctttat cttctaagct tctcacacct tttctctcgt ctctccacga tttacacact
     1861 atttgcaatg aatctcactc tgatgagcgt atctatcaag tccttgtctc ttcttttgac
     1921 aaaacttcaa agaaatcgcc gagcgctact agtgcaggag ctgtaagttt aaaataactt
     1981 cgtatagtat accttatacg aagttatttt cagggagagc aacttaagat ttttgctttc
     2041 gtcgtaaatc tacacacgcg tctcttccgt gcgagagtcc aagccagcag ccaaattcgt
     2101 tgactgagta ttcaacgttt atacgttgtc ggcaacgaga aataggaaaa tgcatcggga
     2161 aatgttcttt tttcgatttt ttccaaggtt ttgacaaatt ttaccacgaa ttttgctatg
     2221 ttttcaatta aaaaatatgt tattcaactg tttctatgag gaaaataagg ctttgcatgt
     2281 aattttctta ttcagcataa tttttaatta atttgaattt tctgtcctaa cgtttatttt
     2341 gttttcttgg ttatgactga tctgaaatta atttttgaat tttaaggtaa tatgaaaaaa
     2401 cccgagttga ccgccacatc cgtagagaag ttcctcatcg agaagttcga ctccgtctcc
     2461 gacctcatgc aactctccga gggagaggag tcccgtgcct tctccttcga cgtcggagga
     2521 cgtggatacg tcctccgtgt caactcctgc gccgacggat tctacaagga ccgttacgtc
     2581 taccgtcact tcgcctccgc cgccctccca atcccagagg tcctcgacat cggagagttc
     2641 tccgagtccc tcacctactg catctcccgt cgtgcccaag gagtcaccct ccaagacctc
     2701 ccagagaccg agctcccagc cgtcctccaa ccagtcgccg aggccatgga cgccatcgcc
     2761 gccgccgacc tctcccaaac ctccggattc ggaccattcg gaccacaagg aatcggacaa
     2821 tacaccacct ggcgtgactt catctgcgcc atcgccgacc cacacgtcta ccactggcaa
     2881 accgtcatgg acgacaccgt ctccgcctcc gtcgcccaag ccctcgacga gctcatgctc
     2941 tgggccgagg actgcccaga ggtccgtcac ctcgtccacg ccgacttcgg atccaacaac
     3001 gtcctcaccg acaacggacg tatcaccgcc gtcatcgact ggtccgaggc catgttcgga
     3061 gactcccaat atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg
     3121 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     3181 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     3241 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     3301 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     3361 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3421 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3481 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     3541 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     3601 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     3661 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     3721 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     3781 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     3841 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     3901 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     3961 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4021 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     4081 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     4141 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctga
//
