LOCUS Exported 5683 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION pBXPHM3 with scaffold Sybody of the loop library. ACCESSION . VERSION . KEYWORDS SB_loop 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5683 /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 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 2570..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 //