LOCUS       Exported                5695 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  pBXPHM3 with scaffold Sybody of the convex library.
ACCESSION   .
VERSION     .
KEYWORDS    SB_convex
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5695)
  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 5695)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5695
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
     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             1978..2007
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2023..2040
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2096..2113)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2266..2352
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2444..2471
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2490..2581
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2582..3442
                     /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(2800..2819)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3613..4201
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4102..4121
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4355..4372
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4387..4527
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4453..4472)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4613..4635
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4791..5669)
                     /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 agtcaagtcc agctggtgga atcgggtggt
     1621 ggtagcgtcc aggcgggtgg tagcctgcgt ctgagctgtg cggctagcgg ctctatttcc
     1681 agcatcacgt acctgggctg gtttcgccag gcaccgggca aagagcgtga gggcgtcgca
     1741 gcgctgagca ccagctccgg taccacctac tacgcggaca gcgttaaggg tcgtttcacg
     1801 gtgagcctgg acaacgccaa gaataccgtg tatctgcaaa tgaacagctt gaaaccggaa
     1861 gatactgctt tgtattactg cgcggcagcc agcagcggct ccagcagccc gctgtctagc
     1921 agcagctata cgtactgggg tcagggcacc caagttaccg tttctgcata ataactagaa
     1981 caaaaactca tctcagaaga ggatctgaat agcgccgtcg accatcatca tcatcatcat
     2041 tgagtttaaa cggtctccag cttggctgtt ttggcggatg agagaagatt ttcagcctga
     2101 tacagattaa atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta
     2161 gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg
     2221 gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag
     2281 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
     2341 agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg
     2401 cgggcaggac gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg
     2461 gatggccttt ttgcgtttct acaaactctt ttgtttattt ttctaaatac attcaaatat
     2521 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     2581 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     2641 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     2701 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     2761 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     2821 ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     2881 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     2941 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     3001 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     3061 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     3121 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     3181 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     3241 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     3301 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     3361 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     3421 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     3481 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     3541 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     3601 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3661 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3721 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     3781 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3841 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3901 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3961 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4021 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4081 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4141 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4201 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4261 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     4321 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     4381 agtgcccctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg
     4441 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
     4501 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
     4561 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     4621 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagca gatcaattcg
     4681 cgcgcgaagg cgaagcggca tgcataatgt gcctgtcaaa tggacgaagc agggattctg
     4741 caaaccctat gctactccgt caagccgtca attgtctgat tcgttaccaa ttatgacaac
     4801 ttgacggcta catcattcac tttttcttca caaccggcac ggaactcgct cgggctggcc
     4861 ccggtgcatt ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga
     4921 ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt
     4981 tggtcctcgc gccagcttaa gacgctaatc cctaactgct ggcggaaaag atgtgacaga
     5041 cgcgacggcg acaagcaaac atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc
     5101 aggtgatcgc tgatgtactg acaagcctcg cgtacccgat tatccatcgg tggatggagc
     5161 gactcgttaa tcgcttccat gcgccgcagt aacaattgct caagcagatt tatcgccagc
     5221 agctccgaat agcgcccttc cccttgcccg gcgttaatga tttgcccaaa caggtcgctg
     5281 aaatgcggct ggtgcgcttc atccgggcga aagaaccccg tattggcaaa tattgacggc
     5341 cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt aaacccactg gtgataccat
     5401 tcgcgagcct ccggatgacg accgtagtga tgaatctctc ctggcgggaa cagcaaaata
     5461 tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg accgcgaatg
     5521 gtgagattga gaatataacc tttcattccc agcggtcggt cgataaaaaa atcgagataa
     5581 ccgttggcct caatcggcgt taaacccgcc accagatggg cattaaacga gtatcccggc
     5641 agcaggggat cattttgcgc ttcagccata cttttcatac tcccgccatt cagag
//
