LOCUS       sequence-66342-e        4737 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SNAP-GFP fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4737)
  AUTHORS   Diamante L, Gatti-Lafranconi P, Schaerli Y, Hollfelder F
  TITLE     In vitro affinity screening of protein and peptide binders by 
            megavalent bead surface display.
  JOURNAL   Protein Eng Des Sel. 2013 Oct;26(10):713-24. doi: 
            10.1093/protein/gzt039. Epub 2013 Aug 26.
  PUBMED    23980186
REFERENCE   2  (bases 1 to 4737)
  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: 
            "10.1093/protein/gzt039"; journalName: "Protein Eng Des Sel"; date: 
            "2013-10"; volume: "26"; issue: "10"; pages: "713-24"
COMMENT     Sequence Label: pIVEX-SNAP-GFP
FEATURES             Location/Qualifiers
     source          1..4737
                     /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     366..388
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     380..397
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     381..397
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     517..536
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     620..639
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        620..638
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             670..692
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1270..1986
                     /codon_start=1
                     /product="GFP variant optimized for excitation by UV light"
                     /label=GFPuv
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1300..1328)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1923..1943
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2040..2058)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2054..2101
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2467..2472,2473..2489,2490..2495))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2516..2532)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2516..2532)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2529..2551)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2540..2556
                     /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(2564..2570,2571..2588,2589..2594))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2609..2630
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2747..2764)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2918..3506)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2998..3017)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3677..4468,4469..4537))
                     /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     4300..4319
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4538..4642)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4710..4728
                     /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 accatatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
      241 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
      301 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      361 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagcttgcat gcaaggagat
      421 ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct
      481 catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc
      541 agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga
      601 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
      661 agaaataatt ttgtttaact ttaagaagga gatatacata tgggcctggg ctccatggac
      721 aaggattgtg aaatgaaacg caccacactg gacagccctt tggggaagct ggagctgtct
      781 ggttgtgagc agggtctgca cgaaataatt tttctgggca aggggacgtc tgcagctgat
      841 gccgtggagg tcccagcccc cgctgcggtt ctcggaggtc cggagcccct gatgcaggct
      901 acagcctggc tgaatgccta tttccaccag cccgaggcta tcgaagagtt ccccgtgccg
      961 gctcttcacc atcccgtttt ccagcaagag tcgttcacca gacaggtgtt atggaagctg
     1021 ctgaaggttg tgaaattcgg agaagtgatt tcttactctc acttagcagc cctggcaggc
     1081 aaccccgcag ccacggcagc agtgaagacg gcactgagtg gcaatcctgt ccctatcctg
     1141 atcccgtgcc acagagtggt ccagggggat ttggatgtgg gcggttacga gggtggactg
     1201 gccgtgaagg aatggcttct ggcccatgaa ggccaccggt tggggaagcc aggcttggga
     1261 agcggccgca tgagtaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa
     1321 ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga aggtgatgca
     1381 acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc tgttccttgg
     1441 ccaacacttg tcactactct gacgtatggt gttcaatgct tttcccgtta tccggatcac
     1501 atgaaacggc atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaacgcact
     1561 atattcttca aagatgacgg gaactacaag acgcgtgctg aagtcaagtt tgaaggtgat
     1621 acccttgtta atcgtatcga gttaaaaggt attgatttta aagaagatgg aaacattctc
     1681 ggacacaaac tcgagtacaa ctataactca cacaatgtat acatcacggc agacaaacaa
     1741 aagaatggaa tcaaagctaa cttcaaaatt cgccacaaca ttgaagatgg ttccgttcaa
     1801 ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac
     1861 aaccattacc tgtcgacaca atctgccctt tcgaaagatc ccaacgaaaa gcgtgaccac
     1921 atggtccttc ttgagtttgt aactgctgct gggattacac atggcatgga tgagctctac
     1981 aaataatgag gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
     2041 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     2101 gctgaaagga ggaactatat ccggatatcc acaggacggg tgtggtcgcc atgatcgcgt
     2161 agtcgatagt ggctccaagt agcgaagcga gcaggactgg gcggcggcca aagcggtcgg
     2221 acagtgctcc gagaacgggt gcgcatagaa attgcatcaa cgcatatagc gctagcagca
     2281 cgccatagtg actggcgatg ctgtcggaat ggacgatatc ccgcaagagg cccggcagta
     2341 ccggcataac caagcctatg cctacagcat ccagggtgac ggtgccgagg atgacgatga
     2401 gcgcattgtt agatttcata cacggtgcct gactgcgtta gcaatttaac tgtgataaac
     2461 taccgcatta aagcttatcg atgataagct gtcaaacatg agaattcgta atcatgtcat
     2521 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     2581 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     2641 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     2701 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     2761 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     2821 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     2881 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     2941 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3001 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3061 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3121 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3181 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3241 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3301 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     3361 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3421 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3481 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3541 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3601 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3661 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3721 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     3781 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     3841 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     3901 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     3961 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4021 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4081 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4141 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4201 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4261 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4321 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4381 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4441 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4501 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     4561 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4621 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     4681 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtc
//
