LOCUS       sequence-211287-        5665 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBXPHM3 with scaffold Sybody of the concave library.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5665)
  AUTHORS   Zimmermann I, Egloff P, Hutter CA, Arnold FM, Stohler P, Bocquet N, 
            Hug MN, Huber S, Siegrist M, Hetemann L, Gera J, Gmur S, Spies P, 
            Gygax D, Geertsma ER, Dawson RJ, Seeger MA
  TITLE     Synthetic single domain antibodies for the conformational trapping 
            of membrane proteins.
  JOURNAL   Elife. 2018 May 24;7. pii: 34317. doi: 10.7554/eLife.34317.
  PUBMED    29792401
REFERENCE   2  (bases 1 to 5665)
  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: "Elife."; 
            date: "2018-05-24"; volume: "7. pii: 34317. doi"; pages: " 
            10.7554/eLife.34317"
COMMENT     Sequence Label: SB_concave
FEATURES             Location/Qualifiers
     source          1..5665
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     sig_peptide     319..384
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     CDS             424..453
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             457..1557
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1531..1554
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1567..1590
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1948..1977
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1993..2010
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2066..2083)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2236..2322
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2414..2441
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2460..2551
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2552..2620,2621..3412)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2770..2789)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3583..4171
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4072..4091
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4325..4342
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4357..4497
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4423..4442)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4583..4605
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4761..5639)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggcact
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aattaaccat gaaatacctg ctgccgaccg ctgctgctgg tctgctgctc
      361 ctcgctgccc agccggcgat ggccatggat atcggaatta attcggatcc gaattcgagc
      421 tcacatcacc atcaccatca ccatcaccat caccccatga aaatcgaaga aggtaaactg
      481 gtaatctgga ttaacggcga taaaggctat aacggtctcg ctgaagtcgg taagaaattc
      541 gagaaagata ccggaattaa agtcaccgtt gagcatccgg ataaactgga agagaaattc
      601 ccacaggttg cggcaactgg cgatggccct gacattatct tctgggcaca cgaccgcttt
      661 ggtggctacg ctcaatctgg cctgttggct gaaatcaccc cggacaaagc gttccaggac
      721 aagctgtatc cgtttacctg ggatgccgta cgttacaacg gcaagctgat tgcttacccg
      781 atcgctgttg aagcgttatc gctgatttat aacaaagatc tgctgccgaa cccgccaaaa
      841 acctgggaag agatcccggc gctggataaa gaactgaaag cgaaaggtaa gagcgcgctg
      901 atgttcaacc tgcaagaacc gtacttcacc tggccgctga ttgctgctga cgggggttat
      961 gcgttcaagt atgaaaacgg caagtacgac attaaagacg tgggcgtgga taacgctggc
     1021 gcgaaagcgg gtctgacctt cctggttgac ctgattaaaa acaaacacat gaatgcagac
     1081 accgattact ccatcgcaga agctgccttt aataaaggcg aaacagcgat gaccatcaac
     1141 ggcccgtggg catggtccaa catcgacacc agcaaagtga attatggtgt aacggtactg
     1201 ccgaccttca agggtcaacc atccaaaccg ttcgttggcg tgctgagcgc aggtattaac
     1261 gccgccagtc cgaacaaaga gctggcaaaa gagttcctcg aaaactatct gctgactgat
     1321 gaaggtctgg aagcggttaa taaagacaaa ccgctgggtg ccgtagcgct gaagtcttac
     1381 gaggaagagt tggcgaaaga tccacgtatt gccgccacta tggaaaacgc ccagaaaggt
     1441 gaaatcatgc cgaacatccc gcagatgtcc gctttctggt atgccgtgcg tactgcggtg
     1501 atcaacgccg ccagcggtcg tcagactgtc gatgaagccc tgaaagacgc gcagactccg
     1561 ggtagcctgg aagttctgtt ccaggggccc agtcaggttc agctggttga gagcggtggt
     1621 ggcctggtcc aagctggcgg ttcgctgcgt ctgagctgcg ccgcaagcgg tttcccggtg
     1681 agcagcagca cgatgacctg gtatcgtcag gcaccgggca aagaacgtga gtgggtcgcg
     1741 gcgatttcca gctctggtag caccacgacc tacgcagatt ctgttaaggg ccgctttacc
     1801 atcagccgcg acaacgcgaa gaatacggtc tatttgcaga tgaatagcct gaaaccggaa
     1861 gataccgcgg tttactactg taccgtgacg gtgggtagca cctacacggg ccaaggtacc
     1921 caagtgactg tgagcgcata ataactagaa caaaaactca tctcagaaga ggatctgaat
     1981 agcgccgtcg accatcatca tcatcatcat tgagtttaaa cggtctccag cttggctgtt
     2041 ttggcggatg agagaagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc
     2101 tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga
     2161 actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat gcgagagtag
     2221 ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     2281 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg
     2341 aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg
     2401 catcaaatta agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactctt
     2461 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     2521 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     2581 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     2641 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     2701 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     2761 taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg
     2821 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     2881 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     2941 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     3001 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     3061 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     3121 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     3181 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     3241 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     3301 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     3361 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     3421 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     3481 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     3541 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3601 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3661 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3721 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3781 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3841 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3901 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3961 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4021 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4081 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4141 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4201 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     4261 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     4321 gcgcagcgag tcagtgagcg aggaagcgga agtgcccctg atgcggtatt ttctccttac
     4381 gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc
     4441 cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc
     4501 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     4561 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     4621 ccgaaacgcg cgaggcagca gatcaattcg cgcgcgaagg cgaagcggca tgcataatgt
     4681 gcctgtcaaa tggacgaagc agggattctg caaaccctat gctactccgt caagccgtca
     4741 attgtctgat tcgttaccaa ttatgacaac ttgacggcta catcattcac tttttcttca
     4801 caaccggcac ggaactcgct cgggctggcc ccggtgcatt ttttaaatac ccgcgagaaa
     4861 tagagttgat cgtcaaaacc aacattgcga ccgacggtgg cgataggcat ccgggtggtg
     4921 ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc gccagcttaa gacgctaatc
     4981 cctaactgct ggcggaaaag atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg
     5041 acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg
     5101 cgtacccgat tatccatcgg tggatggagc gactcgttaa tcgcttccat gcgccgcagt
     5161 aacaattgct caagcagatt tatcgccagc agctccgaat agcgcccttc cccttgcccg
     5221 gcgttaatga tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc atccgggcga
     5281 aagaaccccg tattggcaaa tattgacggc cagttaagcc attcatgcca gtaggcgcgc
     5341 ggacgaaagt aaacccactg gtgataccat tcgcgagcct ccggatgacg accgtagtga
     5401 tgaatctctc ctggcgggaa cagcaaaata tcacccggtc ggcaaacaaa ttctcgtccc
     5461 tgatttttca ccaccccctg accgcgaatg gtgagattga gaatataacc tttcattccc
     5521 agcggtcggt cgataaaaaa atcgagataa ccgttggcct caatcggcgt taaacccgcc
     5581 accagatggg cattaaacga gtatcccggc agcaggggat cattttgcgc ttcagccata
     5641 cttttcatac tcccgccatt cagag
//
