LOCUS       sequence-193139-        2876 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  6XHis::TEV::3XFlag::linker::Myc.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2876)
  AUTHORS   Paix A, Folkmann A, Seydoux G
  TITLE     Precision genome editing using CRISPR-Cas9 and linear repair 
            templates in C. elegans.
  JOURNAL   Methods. 2017 May 15;121-122:86-93. doi: 
            10.1016/j.ymeth.2017.03.023. Epub 2017 Apr 7.
  PUBMED    28392263
REFERENCE   2  (bases 1 to 2876)
  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: "Methods."; 
            date: "2017-05-15"; volume: "121-122:86-93. doi"; pages: " 
            10.1016/j.ymeth.2017.03.023. Epub 2017 Apr 7"
COMMENT     Sequence Label: pAP757-1
FEATURES             Location/Qualifiers
     source          1..2876
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(20..37)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(191..779)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(271..290)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(950..1741,1742..1810))
                     /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     1573..1592
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1811..1915)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1983..2001
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2039..2061)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2161..2180
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2374..2396
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2388..2405
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2389..2405
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2432..2449
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2450..2470
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2471..2536
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2588..2617
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2665..2681)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2665..2681)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2678..2700)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2689..2705
                     /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)."
     promoter        complement(join(2713..2719,2720..2737,2738..2743))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2758..2779
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
       61 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
      121 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
      181 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
      241 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
      301 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
      361 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
      421 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
      481 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
      541 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
      601 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
      661 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
      721 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
      781 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
      841 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
      901 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
      961 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     1021 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     1081 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     1141 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     1201 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     1261 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     1321 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     1381 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     1441 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     1501 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     1561 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     1621 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     1681 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     1741 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     1801 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     1861 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     1921 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     1981 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     2041 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     2101 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     2161 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
     2221 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg
     2281 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     2341 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     2401 ccagtgaatt cgagctcggt acccggggat ccatcatcac catcaccatg aaaatttata
     2461 ttttcagggt gactacaaag accatgacgg tgattataaa gatcatgata tcgattacaa
     2521 ggatgacgat gacaagtgcc cgggggatcg gtggagctcc accggtggcg gccgctctag
     2581 aactagtgaa caaaaactga tatcagaaga ggatctggat cctctagagt cgacctgcag
     2641 gcatgcaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     2701 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     2761 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2821 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgta
//
