PComb3X PComb3X Map Shows Map Of Sequence Available
pComb3X map shows map of sequence available on GenBank, Accession # AF268281Only pComb3XSS, pComb3XTT and pComb3XLambda are available upon MTA request (see below)See last page for sequencing IlB032100Pe00330SfiIHis6Amber Stop 00Igene 00015020001600170018001900pRAmF1iorNheI
pComb3XSS, available for request by MTAContains nonsense SS stuffer in light chain and heavy chain cloning regionsRefer to Protocols 9.1-9.9 in Phage Display: A Laboratory Manual for detail on 2600NheI0IIIgene020270028002900F1o riB 00380011003000Amp RChtStu01004000090LignhaiSpeISfiIHis6Amber Stop CodonainStuffererffer
pComb3XTT, available for request by MTAContains human Fab to Tetanus ToxinIntended for use as template for human kappa light chain and human CH1 heavy chain constantregions, see Protocols 9.4, 9.6 and 9.8 in Phage Display: A Laboratory Manual, can be used alsofor cloning and Fab expression controlpComb3XTTbleRegionSfiIriaOmpALeaderLacZ Promoter0430420ain100Ch0200ht4600300omps erLeadXhoIHeavy Chain Variable RegionGX03300170003101600032015001000HA80Tag3His6 Tag00200021022002300eIIINheISfiIAmber 0804000KX07041000LigCainRntHeavyChainCH1
pComb3XLambda, available for request by MTAContains nonsense antibody with human lambda constant regionIntended for use as template for human lambda light chain and human CH1 heavy chain constantregions, see Protocol 9.1 in Phage Display: A Laboratory ManualpComb3XLambdariableinsertSfiI SitevaOmpALeaderLacZ Promoter004342se200en100300ns4600No44000450LCSacI Site04005000060XbaIPelBerLeadXhoIHeavy Chain Variable Region033000H0ATag180017000310160003201500His6 Tag300200021022002300eIIINheISfiIAmber 007041000dambLanmaHuionegtRntansCoHeavyChainCH1
pComb3XscFv Map just to show what vector looks like with scFv cloned inNo scFv is available for requestNote: depicts short linker, long linker is 12 AApComb3XscFvLacZ PromoterongiReeblriaVaChain4007AA Linker00t30omps e 0q0037500Heahavy C34003000gback2900HA 0210022002300pRAmF1 O r iNheIgeneIIISfiIHis6 Tag Amber Stop12000280SpeI1100dps eq1000310090032008003300070060 rSfiIinVariableRegion
Primers for sequencing Fabs/scFv in pComb3 vectors:ompseq – sequences from ompA leader region forward through light chainAAG ACA GCT ATC GCG ATT GCA Gkpel – primes from pelB leader sequence backward thorugh light chainCGG CTG CCG TAG GCA ATA GGTKX – can be used with chimeric or human Fab to sequence backward through light chain variable region (primes atbeginning of constant region)GAA GTT ATT CAG CAG GCA CACpelseq – primes from pelB leader sequence forward through heavy chainACC TAT TGC CTA CGG CAG CCGgback – primes from geneIII region backward through heavy chainGCC CCC TTA TTA GCG TTT GCC ATCdpseq – primes from HA tag (only in pComb3X) back through heavy chainAGA AGC GTA GTC CGG AAC GTCGX - can be used with chimeric or human Fab to sequence backward through heavy chain variable region (primes atbeginning of constant region)GGG AAG TAG TCC TTG ACC AGG CHRML-F – can be used in scFV to sequence forward from the linker through the heavy chainGGT GGT TCC TCT AGA TCT TCCHRML-B – can be used in scFV to sequence backward from the linker through the light chainGGA AGA TCT AGA GGA ACC ACC
pComb3X map shows map of sequence available on GenBank, Accession # AF26 8281 Only pComb3XSS, pComb3XTT and pComb3XLambda are available upon MTA
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