LOCUS       sequence-130626-        3539 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses myc::TEV::mTagGFP::TEV::3Xflag in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3539)
  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 3539)
  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: AP588-1
FEATURES             Location/Qualifiers
     source          1..3539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             422..451
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             452..472
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             473..1183
                     /codon_start=1
                     /product="monomeric green fluorescent protein, also known 
                     as mTagGFP"
                     /label=TagGFP2
                     /note="mammalian codon-optimized"
                     /translation="SGGEELFAGIVPVLIELDGDVHGHKFSVRGEGEGDADYGKLEIKF
                     ICTTGKLPVPWPTLVTTLCYGIQCFARYPEHMKMNDFFKSAMPEGYIQERTIQFQDDGK
                     YKTRGEVKFEGDTLVNRIELKGKDFKEDGNILGHKLEYSFNSHNVYIRPDKANNGLEAN
                     FKTRHNIEGGGVQLADHYQTNVPLGDGPVLIPINHYLSTQTKISKDRNEARDHMVLLES
                     FSACCHTHGMDELYR"
     CDS             1184..1204
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1205..1270
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1318..1334)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1318..1334)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1331..1353)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1342..1358
                     /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(1366..1372,1373..1390,1391..1396))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1411..1432
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1549..1566)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1720..2308)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1800..1819)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2479..3270,3271..3339))
                     /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     3102..3121
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3340..3444)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3512..3530
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acccggggat
      421 cgaacaaaaa ctgatatcag aagaggatct ggaaaattta tattttcagg gtagcggggg
      481 cgaggagctg ttcgccggca tcgtgcccgt gctgatcgag ctggacggcg acgtccacgg
      541 acacaagttc tccgtccgcg gagagggaga gggagacgcc gactacggaa agctggagat
      601 caagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctag tgaccaccct
      661 ctgctacggc atccagtgct tcgcccgcta ccccgagcac atgaagatga acgacttctt
      721 caagagcgcc atgcccgagg gctacataca ggagcgcacc atccagttcc aggacgacgg
      781 caagtacaag acccgcggcg aggtgaagtt cgagggcgac accctggtga accgcataga
      841 gctgaagggc aaggacttca aggaggacgg caacatcctg ggccacaagc tggagtacag
      901 cttcaacagc cacaatgtgt acatccgccc cgacaaggcc aacaacggcc tggaggctaa
      961 cttcaagacc cgccacaaca tcgagggcgg cggcgtgcag ctagccgacc actaccagac
     1021 caacgtgccc ctgggcgacg gccccgtgct aatccccatc aaccactacc tgagcactca
     1081 gaccaagatc agcaaggatc gcaacgaggc ccgcgaccac atggtgctcc tggagtcctt
     1141 cagcgcctgc tgccacaccc acggcatgga cgagctgtac agggagaacc tctacttcca
     1201 aggagactac aaagaccatg acggtgatta taaagatcat gatatcgatt acaaggatga
     1261 cgatgacaag gatcctctag agtcgacctg caggcatgca agcttggcgt aatcatggtc
     1321 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     1381 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     1441 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     1501 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     1561 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     1621 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     1681 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1741 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1801 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1861 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1921 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1981 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2041 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2101 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2161 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2221 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2281 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2341 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2401 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2461 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2521 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     2581 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     2641 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     2701 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     2761 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     2821 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     2881 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     2941 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     3001 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     3061 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     3121 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     3181 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     3241 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     3301 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     3361 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     3421 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     3481 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtc
//
