LOCUS       sequence-431860-        4379 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty repair plasmid for ChrII split hygromycinR landing pad.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4379)
  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 4379)
  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: pMS178
FEATURES             Location/Qualifiers
     source          1..4379
                     /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(2162..2195)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(3318..3337)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3437..3459
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3497..3515)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3583..3687
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3688..3756,3757..4379,1..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(3906..3925)
                     /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 ccaagctcac tcgtttaggc tattccccta tttttgatat
     1381 tccttcgcac atatgaaaac tacttttttt cgaaactgtt aactccagaa tttataaatc
     1441 tatagccctt acttgattat ttattatcat ggttactcac ctcatgtctc tttctctttt
     1501 caagaagtct tcgttcatat ctgttgtctc tttcctcctt ttgaatctct gctttgctct
     1561 tcgccattgt ttcctgaaaa taatgtaact tgaattgtgt aatatacttt tttaatttga
     1621 atttggcttg taacgcggaa tcactacgtg cgggatcatt tcttactaga aaacccagaa
     1681 aatgccatat ttcacttatc tcggggtcat ttctaattag aaaagctaca aaaaccattt
     1741 ctaattacgg tgactttaga actggtcgaa atagctaatt agaaatgggt ttttgctaat
     1801 tagaggtgac tctatgttat ctgagtaagc ctcgtttgta tgtaaactca cagtactaat
     1861 aaatgcaaga cacccgggtt tgtctagata tgaaataatt gaaatatcaa ttctgacaga
     1921 caataatggt aatcttgata aggagttcca cgcccaggag aacacgttag ttttcttgtt
     1981 tttgattgcg tgcgttattt tggagaaaaa ctcgattttt tacaaaataa ttttttgaaa
     2041 ggaacactgt tcaataagtt ttgtcttttt tctcagttgt gatacggttt tttattcttt
     2101 tttgtagtta tacagaagac cgcctgcagg cttaagttca atccggaaag gtaagtttaa
     2161 aataacttcg tatagcatac attatacgaa gttattttca gggagagcaa aagatttttg
     2221 ctttcgtcgt aaatctacac acgcgtctct tccgtgcgag agtccaagcc agcagccaaa
     2281 ttcgttgact gagtattcaa cgtttatacg ttgtcggcaa cgagaaatag gaaaatgcat
     2341 cgggaaatgt tcttttttcg attttttcca aggttttgac aaattttacc acgaattttg
     2401 ctatgttttc aattaaaaaa tatgttattc aactgtttct atgaggaaaa taaggctttg
     2461 catgtaattt tcttattcag cataattttt aattaatttg aattttctgt cctaacgttt
     2521 attttgtttt cttggttatg actgatctga aattaatttt tgaattttaa ggtaatatga
     2581 aaaaacccga gttgaccgcc acatccgtag agaagttcct catcgagaag ttcgactccg
     2641 tctccgacct catgcaactc tccgagggag aggagtcccg tgccttctcc ttcgacgtcg
     2701 gaggacgtgg atacgtcctc cgtgtcaact cctgcgccga cggattctac aaggaccgtt
     2761 acgtctaccg tcacttcgcc tccgccgccc tcccaatccc agaggtcctc gacatcggag
     2821 agttctccga gtccctcacc tactgcatct cccgtcgtgc ccaaggagtc accctccaag
     2881 acctcccaga gaccgagctc ccagccgtcc tccaaccagt cgccgaggcc atggacgcca
     2941 tcgccgccgc cgacctctcc caaacctccg gattcggacc attcggacca caaggaatcg
     3001 gacaatacac cacctggcgt gacttcatct gcgccatcgc cgacccacac gtctaccact
     3061 ggcaaaccgt catggacgac accgtctccg cctccgtcgc ccaagccctc gacgagctca
     3121 tgctctgggc cgaggactgc ccagaggtcc gtcacctcgt ccacgccgac ttcggatcca
     3181 acaacgtcct caccgacaac ggacgtatca ccgccgtcat cgactggtcc gaggccatgt
     3241 tcggagactc ccaatatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     3301 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     3361 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3421 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     3481 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     3541 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3601 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3661 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     3721 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     3781 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     3841 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     3901 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     3961 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4021 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4081 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4141 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4201 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     4261 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4321 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctga
//
