LOCUS       sequence-211298-        5683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBXPHM3 with scaffold Sybody of the loop library.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5683)
  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 5683)
  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_loop
FEATURES             Location/Qualifiers
     source          1..5683
                     /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             1966..1995
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2011..2028
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2084..2101)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2254..2340
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2432..2459
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2478..2569
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2570..2638,2639..3430)
                     /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(2788..2807)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3601..4189
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4090..4109
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4343..4360
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4375..4515
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4441..4460)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4601..4623
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4779..5657)
                     /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 taacgtgaaa gacagcggta gctccagcag ctcctacgac
     1921 tattggggcc aaggtaccca agtgactgtg agcgcataat aactagaaca aaaactcatc
     1981 tcagaagagg atctgaatag cgccgtcgac catcatcatc atcatcattg agtttaaacg
     2041 gtctccagct tggctgtttt ggcggatgag agaagatttt cagcctgata cagattaaat
     2101 cagaacgcag aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc
     2161 cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt
     2221 ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa
     2281 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
     2341 ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc
     2401 ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt
     2461 gcgtttctac aaactctttt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2521 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     2581 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     2641 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     2701 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     2761 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc
     2821 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     2881 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     2941 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     3001 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     3061 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     3121 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     3181 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     3241 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     3301 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     3361 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     3421 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     3481 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     3541 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3601 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3661 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3721 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     3781 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3841 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3901 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3961 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4021 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4081 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4141 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4201 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     4261 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     4321 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag tgcccctgat
     4381 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag
     4441 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     4501 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     4561 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     4621 aggttttcac cgtcatcacc gaaacgcgcg aggcagcaga tcaattcgcg cgcgaaggcg
     4681 aagcggcatg cataatgtgc ctgtcaaatg gacgaagcag ggattctgca aaccctatgc
     4741 tactccgtca agccgtcaat tgtctgattc gttaccaatt atgacaactt gacggctaca
     4801 tcattcactt tttcttcaca accggcacgg aactcgctcg ggctggcccc ggtgcatttt
     4861 ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg
     4921 ataggcatcc gggtggtgct caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc
     4981 cagcttaaga cgctaatccc taactgctgg cggaaaagat gtgacagacg cgacggcgac
     5041 aagcaaacat gctgtgcgac gctggcgata tcaaaattgc tgtctgccag gtgatcgctg
     5101 atgtactgac aagcctcgcg tacccgatta tccatcggtg gatggagcga ctcgttaatc
     5161 gcttccatgc gccgcagtaa caattgctca agcagattta tcgccagcag ctccgaatag
     5221 cgcccttccc cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg
     5281 tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca gttaagccat
     5341 tcatgccagt aggcgcgcgg acgaaagtaa acccactggt gataccattc gcgagcctcc
     5401 ggatgacgac cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc acccggtcgg
     5461 caaacaaatt ctcgtccctg atttttcacc accccctgac cgcgaatggt gagattgaga
     5521 atataacctt tcattcccag cggtcggtcg ataaaaaaat cgagataacc gttggcctca
     5581 atcggcgtta aacccgccac cagatgggca ttaaacgagt atcccggcag caggggatca
     5641 ttttgcgctt cagccatact tttcatactc ccgccattca gag
//
