LOCUS       sequence-204277-        3026 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian GFP11 with 
            linker::TEV::3xFlag::Ollas::Myc::V5::HAtag::other linker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3026)
  AUTHORS   Paix A, Folkmann A, Goldman DH, Kulaga H, Grzelak MJ, Rasoloson D, 
            Paidemarry S, Green R, Reed RR, Seydoux G
  TITLE     Precision genome editing using synthesis-dependent repair of 
            Cas9-induced DNA breaks.
  JOURNAL   Proc Natl Acad Sci U S A. 2017 Nov 28. pii: 1711979114. doi: 
            10.1073/pnas.1711979114.
  PUBMED    29183983
REFERENCE   2  (bases 1 to 3026)
  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: "Proc Natl 
            Acad Sci U S A. 2017 Nov 28. pii: 1711979114. doi: 
            10.1073/pnas.1711979114."
COMMENT     Sequence Label: pAP2050-2
FEATURES             Location/Qualifiers
     source          1..3026
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     105..124
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     318..340
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     332..349
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     333..349
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             376..423
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             433..453
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             454..519
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             520..561
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             562..591
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             592..633
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             634..660
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(759..775)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(759..775)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(772..794)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    783..799
                     /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(807..813,814..831,832..837))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    852..873
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(990..1007)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1161..1749)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1241..1260)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1920..2711,2712..2780))
                     /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     2543..2562
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2781..2885)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2953..2971
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(join(3009..3026,1..5))
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
       61 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
      121 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
      181 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
      241 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
      301 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattcgagct
      361 cggtacccgg ggatccgtga ccacatggtc cttcatgagt atgtaaatgc tgctgggatt
      421 acaggtggcg gcgagaacct ttattttcag ggagactata aggaccatga cggggattat
      481 aaagatcacg acattgatta caaggacgac gacgataagt ccggcttcgc taacgaattg
      541 gggccaaggc tcatgggcaa ggaacagaaa ttgatctctg aggaggacct gggtaaacct
      601 atcccaaatc ccctgttggg tttggactca acttacccct acgacgttcc tgactatgct
      661 tgccctggtg accgctggag ttcaacaaga agccgcagat caagaacgag tgatcctcta
      721 gagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt tcctgtgtga
      781 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
      841 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
      901 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
      961 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     1021 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     1081 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     1141 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     1201 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     1261 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     1321 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     1381 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     1441 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     1501 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     1561 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     1621 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     1681 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     1741 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     1801 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     1861 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     1921 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     1981 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     2041 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     2101 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     2161 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     2221 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     2281 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     2341 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     2401 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     2461 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     2521 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     2581 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     2641 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     2701 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     2761 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     2821 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     2881 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     2941 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     3001 ggtgaaaacc tctgacacat gcagct
//
