LOCUS       sequence-193117-        3611 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Myc::TEV::3XFlag::Dendra2::3XFlag::TEV::Myc.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3611)
  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 3611)
  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: pAP635-2
FEATURES             Location/Qualifiers
     source          1..3611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     132..151
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     385..402
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    519..540
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(555..560,561..578,579..585)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    593..609
                     /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     598..620
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     617..633
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     617..633
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(681..710)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(711..731)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(732..797)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(1485..1550)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(1551..1571)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(1572..1601)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1628..1645)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1628..1644)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1637..1659)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1853..1872)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1972..1994
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2032..2050)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2118..2222
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2223..2291,2292..3083)
                     /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(2441..2460)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3254..3611,1..231)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
       61 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      121 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      181 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      241 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      301 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
      361 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
      421 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
      481 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
      541 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
      601 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct tgcatgcctg
      661 caggtcgact ctagaggatc cagatcctct tctgatatca gtttttgttc tccttggaag
      721 tagaggttct ccttgtcatc gtcatccttg taatcgatat catgatcttt ataatcaccg
      781 tcatggtctt tgtagtccca gacttgggat gggagtgggg agtaacgggc gacggcgtgc
      841 tcgtagagct tgaccttgtt gtagtcggag tcgtttccga ggatctcgat acggtggtcg
      901 acgaagtggg cgtctgggag ttggacgacc ttcttggcct tgtaggtggt cttgaagtcg
      961 cagaggtagt gtcctcctcc ctcgaggagg agggccatgt tgatgtttcc gacgaggagt
     1021 ccgtcacgga cgtggagctt ctcggtggat ggctcccact tgagggtctt cttttgcatg
     1081 actggtccgt ttggtgggaa gttggttccc ttgaaacgga cgttttggaa gaagcagtct
     1141 ccctcgaggg agatgtcgga acggatggtg cagattccct tgtcctcgaa ggtcatggta
     1201 cgctcccagg agtatccctc tgggaaggat tgcttgaagt agtctgggat gtcctctggg
     1261 tacttggtga agacacggtt tccgtagtgg acggcggtgg tgaggatgtc gtaggagaat
     1321 gggagtgggg ctccctcctt gacggtgagg ttggcggttt gggttccctc gtatggcttt
     1381 ccctttccct ctccctcgat gacgaaggcg tgtccgttga cgtttccctc catgtggacc
     1441 ttgacacgca tgtcctcctt gatgaggttg attcctgggg tgttcttgtc gtcgtcgtcc
     1501 ttgtagtcga tgtcgtggtc cttgtagtct ccgtcgtggt ccttgtagtc accctgaaaa
     1561 tataaatttt cgaggtcctc ctcggagatg agcttttgct cgatccccgg gtaccgagct
     1621 cgaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     1681 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     1741 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt
     1801 ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct
     1861 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     1921 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     1981 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     2041 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     2101 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     2161 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     2221 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     2281 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     2341 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     2401 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     2461 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     2521 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     2581 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     2641 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     2701 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     2761 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     2821 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     2881 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     2941 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     3001 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     3061 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     3121 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     3181 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     3241 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3301 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3361 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     3421 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3481 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3541 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3601 tggagcgaac g
//
