LOCUS Exported 4642 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION E.coli entry and expression vector for FX cloning system, N-terminal pelB signal sequence and C-terminal myc and 6xHisTag . ACCESSION . VERSION . KEYWORDS pSBinit SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4642) 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 4642) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4642 /organism="synthetic DNA construct" /mol_type="other DNA" sig_peptide 7..69 /gene="pelB (fragment)" /product="leader peptide for secretion" /label=pelB signal sequence CDS complement(120..425) /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /label=ccdB /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" primer_bind complement(222..241) /label=ccdB-fwd /note="ccdB gene, forward primer" CDS 755..784 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 800..817 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(873..890) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" promoter 1154..1256 /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" CDS 1257..1916 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(1323..1342) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" rep_origin 2242..2830 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2731..2750 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2984..3001 /label=L4440 /note="L4440 vector, forward primer" misc_feature 3016..3156 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(3082..3101) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3242..3264 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(3420..4298) /codon_start=1 /gene="araC" /product="L-arabinose regulatory protein" /label=araC /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE EKVNDVAVKLS" promoter 4325..4609 /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 4532..4551 /label=pBAD Forward /note="For vectors with E. coli araBAD promoter, forward primer" ORIGIN 1 atgagtaaat atctgctgcc gaccgcagca gcgggtctgc tgctgctggc agcccagcct 61 gcaatggccg gctcttcaag tcgacctgca gactggctgt gtataaggga gcctgacatt 121 tatattcccc agaacatcag gttaatggcg tttttgatgt cattttcgcg gtggctgaga 181 tcagccactt cttccccgat aacggagacc ggcacactgg ccatatcggt ggtcatcatg 241 cgccagcttt catccccgat atgcaccacc gggtaaagtt cacgggagac tttatctgac 301 agcagacgtg cactggccag ggggatcacc atccgtcgcc cgggcgtgtc aataatatca 361 ctctgtacat ccacaaacag acgataacgg ctctctcttt tataggtgta aaccttaaac 421 tgcatttcac cagcccctgt tctcgtcagc aaaagagccg ttcatttcaa taaaccgggc 481 gacctcagcc atcccttcct gattttccgc tttccagcgt tcggcacgca gacgacgggc 541 ttcattctgc atggttgtgc ttaccagacc ggagatattg acatcatata tgccttgagc 601 aactgatagc tgtcgctgtc aactgtcact gtaatacgct gcttcatagc atacctcttt 661 ttgacatact tcgggtatac atatcagtat atattcttat accgcaaaaa tcagcgcgca 721 aatacgcata ctgttatctg caggaagagc tggcgaacaa aaactcatct cagaagagga 781 tctgaatagc gccgtcgacc atcatcatca tcatcattga gtttaaacgg tctccagctt 841 ggctgttttg gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga 901 agcggtctga taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc 961 atgccgaact cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgaa 1021 ttccccgcgc gcgatgccct ttcgtcttcg aataaatacc tgtgacggaa gatcacttcg 1081 cagaataaat aaatcctggt gtccctgttg ataccgggaa gccctgggcc aacttttggc 1141 gaaaatgaga cgttgatcgg cacgtaagag gttccaactt tcaccataat gaaataagat 1201 cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatgg 1261 agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt 1321 ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta 1381 cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca 1441 ttcttgcccg cctgatgaat gctcatccgg aatttcgtat ggcaatgaaa gacggtgagc 1501 tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt 1561 tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc 1621 aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata 1681 tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcga 1741 atatggacaa cttcttcgcc cccgttttca ctatgggcaa atattatacg caaggcgaca 1801 aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg 1861 gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatttt 1921 tttaaggcag ttattggtgc ccttaaacgc ctggttgcta cgcctgaata agtgataata 1981 agcggatgaa tggcagaaat tcgagcccgc ctaatgagcg ggcttttttt tagcccgcct 2041 aatgagcggg cttttttttc gaaagcaaat tcgacccatc gcgcgcgggg agtcaactca 2101 gcaaaagttc gatttattca acaaagccac gttgtgtctc aaaatcgctg atgttacatt 2161 gcacctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt 2221 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca 2281 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct 2341 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta 2401 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct 2461 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc 2521 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca 2581 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga 2641 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg 2701 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt 2761 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag 2821 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt 2881 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt 2941 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga 3001 ggaagcggaa gtgcccctga tgcggtattt tctccttacg catctgtgcg gtatttcaca 3061 ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac 3121 actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct 3181 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc 3241 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagcag 3301 atcaattcgc gcgcgaaggc gaagcggcat gcataatgtg cctgtcaaat ggacgaagca 3361 gggattctgc aaaccctatg ctactccgtc aagccgtcaa ttgtctgatt cgttaccaat 3421 tatgacaact tgacggctac atcattcact ttttcttcac aaccggcacg gaactcgctc 3481 gggctggccc cggtgcattt tttaaatacc cgcgagaaat agagttgatc gtcaaaacca 3541 acattgcgac cgacggtggc gataggcatc cgggtggtgc tcaaaagcag cttcgcctgg 3601 ctgatacgtt ggtcctcgcg ccagcttaag acgctaatcc ctaactgctg gcggaaaaga 3661 tgtgacagac gcgacggcga caagcaaaca tgctgtgcga cgctggcgat atcaaaattg 3721 ctgtctgcca ggtgatcgct gatgtactga caagcctcgc gtacccgatt atccatcggt 3781 ggatggagcg actcgttaat cgcttccatg cgccgcagta acaattgctc aagcagattt 3841 atcgccagca gctccgaata gcgcccttcc ccttgcccgg cgttaatgat ttgcccaaac 3901 aggtcgctga aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt attggcaaat 3961 attgacggcc agttaagcca ttcatgccag taggcgcgcg gacgaaagta aacccactgg 4021 tgataccatt cgcgagcctc cggatgacga ccgtagtgat gaatctctcc tggcgggaac 4081 agcaaaatat cacccggtcg gcaaacaaat tctcgtccct gatttttcac caccccctga 4141 ccgcgaatgg tgagattgag aatataacct ttcattccca gcggtcggtc gataaaaaaa 4201 tcgagataac cgttggcctc aatcggcgtt aaacccgcca ccagatgggc attaaacgag 4261 tatcccggca gcaggggatc attttgcgct tcagccatac ttttcatact cccgccattc 4321 agagaagaaa ccaattgtcc atattgcatc agacattgcc gtcactgcgt cttttactgg 4381 cacttctcgc taaccaaacc ggtaaccccg cttattaaaa gcattctgta acaaagcggg 4441 accaaagcca tgacaaaaac gcgtaacaaa agtgtctata atcacggcag aaaagtccac 4501 attgattatt tgcacggcgt cacactttgc tatgccatag catttttatc cataagatta 4561 gcggatccta cctgacgctt tttatcgcaa ctctctactg tttctccata cccgtttttt 4621 gggctaacag gaggaattaa cc //