LOCUS       sequence-127878-        3896 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Ollas::mCherry::linker::H2B::V5 in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3896)
  AUTHORS   Paix A, Folkmann A, Rasoloson D, Seydoux G
  TITLE     High Efficiency, Homology-Directed Genome Editing in Caenorhabditis 
            elegans Using CRISPR-Cas9 Ribonucleoprotein Complexes.
  JOURNAL   Genetics. 2015 Sep;201(1):47-54. doi: 10.1534/genetics.115.179382. 
            Epub 2015 Jul 17.
  PUBMED    26187122
REFERENCE   2  (bases 1 to 3896)
  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"; doi: 
            "10.1534/genetics.115.179382"; journalName: "Genetics"; date: 
            "2015-09"; volume: "201"; issue: "1"; pages: "47-54"
COMMENT     Sequence Label: AP582-2
FEATURES             Location/Qualifiers
     source          1..3896
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(142..147,148..165,166..172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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     185..207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     204..220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(268..309)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(1432..1473)
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     primer_bind     complement(1500..1517)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1500..1516)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1509..1531)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1725..1744)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1844..1866
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1904..1922)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1990..2094
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2095..2163,2164..2955)
                     /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(2313..2332)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3126..3714
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3615..3634
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3868..3885
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
      241 atgcctgcag gtcgactcta gaggatcagt actatctaga ccgaggagag ggttagggat
      301 aggcttaccc ttgctggaag tgtacttggt gacggccttg gttccctcag acacggcgtg
      361 cttggcaagt tctcctggga gaatcaaacg gacagcggtt tgaatttcgc gggatgagat
      421 cgttgagcgt ttgttgtaat gagcaagacg ggaagcttcc gaagcgatgc gttcgaatac
      481 atcgttgacg aaggagttca tgatagacat ggccttggag gagactccgg tgtctgggtg
      541 aacttgcttg agaacacggt agatgtagac ggagtacgat tccttgcggg catgacgtct
      601 cttctttccg tcctttggct tggcaacgac ggtcttggcg gccttcttgg ctcccttggc
      661 agatggcttt ggtggactag ttctagagcg gccgccaccg gtggagctcc accgatcccc
      721 cgggcactta tacaattcat ccatgccacc tgtcgagtgc cgcccttcag ctctttcata
      781 ttgctcaacg atagtgtaat cctcgttgtg ggaggtgata tccagtttaa tattaacatt
      841 gtacgctcct ggcagctgaa ctggtttttt ggctttgtaa gttgtcttca cctcagcatc
      901 gtaatgtcct ccatctttca atttaagacg ttgtttaatc tctcccttaa gagcaccgtc
      961 ttcagggtac attctttcac ttgaagcttc ccatcccata gtcttcttct gcataacagg
     1021 tccatccgag gggaaattcg ttcctcgaag tttgacttta taaataaatt ctccatcttg
     1081 gaggctggag tcttgcgtca ctgtaacaac tcctccgtct tcaaaattca ttacgcgttc
     1141 ccacttaaaa ccctccggga atgaaagttt caaataatct ggtatatctg ccggatgttt
     1201 aacataagct ttacttccat acatgaactg tggagagagg atgtcccagg cgaatggtaa
     1261 tggtccgcct ttagttacct ttagttttgc ggtttgtgtt ccctcatatg gtcggccttc
     1321 tccttcacct tcaatttcaa actcatgccc attgacagat ccctccatat gcaccttgaa
     1381 acgcataaac tctttaataa ttgccatgtt atcttcttca ccctttgaga cctttcccat
     1441 gagacgtggt ccgagctcgt tggcgaatcc ggagatcccc gggtaccgag ctcgaattca
     1501 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     1561 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     1621 ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt
     1681 acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat
     1741 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     1801 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     1861 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     1921 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     1981 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     2041 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     2101 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     2161 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     2221 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     2281 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     2341 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     2401 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     2461 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     2521 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     2581 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     2641 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     2701 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     2761 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     2821 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     2881 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     2941 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     3001 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     3061 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3121 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3181 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3241 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     3301 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3361 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3421 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3481 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     3541 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3601 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3661 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3721 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     3781 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     3841 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaaga
//
