LOCUS       sequence-127887-        3584 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TEV::eGFP::myc::3Xflag in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3584)
  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 3584)
  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: AP682-1
FEATURES             Location/Qualifiers
     source          1..3584
                     /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..333)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(385..414)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(join(427..1137,1138..1140))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(467..487)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     1082..1110
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     CDS             complement(1141..1161)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1188..1205)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1188..1204)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1197..1219)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1413..1432)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1532..1554
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1592..1610)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1678..1782
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1783..1851,1852..2643)
                     /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(2001..2020)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2814..3402
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3303..3322
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3556..3573
                     /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 gaggatcctt gtcatcgtca tccttgtaat cgatatcatg
      301 atctttataa tcaccgtcat ggtctttgta gtcactagtt ctagagcggc cgccaccggt
      361 ggagctccac cgatcccccg ggcacagatc ctcttctgat atcagttttt gttcgccggc
      421 tacgtacttg tacagctcgt ccatgccgag tgtaatccca gcagctgtta caaactcaag
      481 aaggaccatg tggtctctct tttcgttggg atctttcgaa agggcagatt gtgtggacag
      541 gtaatggttg tctggtaaaa ggacagggcc atcgccaatt ggagtatttt gttgataatg
      601 gtctgctagt tgaacgcttc catcttcaat gttgtgtcta attttgaagt taactttgat
      661 tccattcttt tgtttgtctg ccatgatgta tacattgtgt gagttatagt tgtattccaa
      721 tttgtgtcca agaatgtttc catcttcttt aaaatcaata ccttttaact cgattctatt
      781 aacaagggta tcaccttcaa acttgacttc agcacgtgtc ttgtagttcc cgtcatcttt
      841 gaaaaatata gttctttcct gtacataacc ttcgggcatg gcactcttga aaaagtcatg
      901 ctgcttcatg tggtcggggt accggctgaa gcactgcacg ccgtaggtca gagtagtgac
      961 aagtgttggc catggaacag gtagttttcc agtagtgcaa ataaatttaa gggtaagttt
     1021 tccgtatgtt gcatcacctt caccctctcc actgacagaa aatttgtgcc cattaacatc
     1081 accatctaat tcaacaagaa ttgggacaac tccagtgaaa agttcttctc ctttactcac
     1141 tccttggaag tagaggttct cgatccccgg gtaccgagct cgaattcact ggccgtcgtt
     1201 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     1261 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     1321 ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc
     1381 ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     1441 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     1501 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     1561 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     1621 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     1681 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     1741 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     1801 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     1861 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     1921 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     1981 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     2041 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     2101 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     2161 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     2221 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     2281 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     2341 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     2401 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     2461 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     2521 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     2581 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     2641 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     2701 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     2761 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2821 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2881 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2941 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     3001 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3061 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3121 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3181 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3241 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3301 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3361 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3421 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3481 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3541 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aaga
//
