# 1 "/u/exp/stan/nmr/lists/pp/hncacbf.wfu" ;hncacbf.fa ;3D HNCACB, gradient enhanced with Watergate reverse INEPT (3-9-19), ; minimal H2O saturation and optional 2H decoupling ;Yamazaki et al., J. Am. Chem. Soc. 1994, 116, 11655-11666 ;Wittekind and Mueller, J. Magn. Reson. B 101 201-205 (1993) ;Mori et al., J. Magn. Reson. B 108, 94-98 (1995) ;Bruker Avance/Xwin-nmr version ;Written up by F. Abildgaard, NMRFAM (abild@nmrfam.wisc.edu) ; ; $Id: pulseprogram,v 1.1 2021/02/10 21:52:55 bmrbsvc Exp $ ; ; Disclaimer: This pulse program is provided "as is" for your ; information. Support for the use of this pulse program is only ; provided to users of the National Magnetic Resonance Facility ; at Madison (NMRFAM). Users of this pulse program employ it at ; their own risk. Neither NMRFAM nor University of Wisconsin-Madison ; are liable for any physical or other damage incurred during the ; use of this pulse program. ; ;f1: 1H, f2: 15N, f3: 13C, f4: 2H ; o1p: 4.7 ppm (H) ; o2p: 118 ppm (N) ; o3p: 43 ppm (CAB) ; o4p: 4 ppm (2H) ; ; N15 evolution, SW=1/(2*in10): [TD(2) = 2*l4] (3d) ; [in30=in10] ; C13 evolution, SW=1/(2*in0): [TD(1) = 2*l6] (3d) ; [sw(1) ~60-70 ppm] ; cnst0=0 (1/2 dwell in C, phase correct is 90 -180) ; cnst0=1 (3/2 dwell in C, phase correct is 270 -540) ; ; ; (optimize over range; leave other term fixed) ; d1 relaxation delay [1-3s] ; d3: set d3 to 3.5ms for Ca and Cb, or 6.8ms for Cb only. ; d21: 1/(2*dNu), dNu=Nu(NH)-Nu(H2O) (watergate, typically 250u) ; pl0=120db ; ; 1H: ; p1 90 H at pl1 [15-19u @ 2db] ; flipback if SHAPED not defined: ; p10 90 H at pl10 [~2.0m @ 42db] (rectangle) ; flipback if SHAPED defined: ; p10 90 H at sp1 [~2m @ ??] ; [spnam1=Seduce.100 or fasqua.256] ; [spoffs1=0] ; (adjust phcor21 to correct phase change) ; ; 15N: ; p2 90 N at pl2 [45-50u @ 2db] ; ; 13C: ; p3 90 CAB at pl3 [14-16u @ 0-2db] ; ; ; 1H decoupling: waltz16 ; pcpd1 90 H at pl11 [96u @ 16-18db] (10 kHz decoupling field) ; [p11=pcpd1; cpdprg1=waltz16] ; 2H decoupling: waltz16 ; pcpd5 at pl15 [550u @ 13.5db] ; [p25=pcpd5; cpdprg5=waltz16] ; 15N decoupling: ; pcpd2 at pl12 [480u @ 22db] ; [cpdprg2=waltz16] ; 13C decoupling: ; pcpd3 at sp3 [306u @ 13-14db] ; 13C' decoupling: compensated seduce (bilobal shifted, +/- 19.6 kHz) ; pcpd3 90 C seduce at sp3 [306u @ 13-14db] ; (equal to a 71.9us rect. 90) ; [cpdprg3=waltz16sp3] ; [spnam3=seduce1c6; spoffs3=0] ;Gradient program: fahncacbf ;ns=8, 16, ..., ds=8, 16,... ; ;Recommendations for gradients: ;p15=500u ;p16=1.0m ;p17=700u ;p18=1.0m ;p19=600u ;p20=100u ;cnst21=6% (4G/cm z) ;cnst22=15% (10G/cm z) ;cnst23=20% (40G/cm combined x,z) ;cnst24=-8% (-5G/cm z) ;cnst25=15% (10G/cm z) ;cnst26=15% (10G/cm z) ;cnst27=12% (8G/cm z) ;cnst28=29% (20G/cm z) ;cnst29=44% (30G/cm z) ;cnst30=1.800 (adjust) for magic angle gradient (Gx/Gz) or 0 for Gz only. ; ;Define one or more of the following options to tailor this pulse program ; to your specific needs. ; ;#define ONE_D ; uncomment for 1D experiment ;#define SHAPED ; uncomment if shaped flip-back pulse ;#define H2_DEC ; uncomment to enable H2 decoupling ;#define SIDECHN ; uncomment for delays optimized for side-chain amide ; ; correlations ; ; the correct expt time (works with XWIN-NMR 2.x) ; ;Define channel assignments: ; ;You shouldn't have to worry about anything beyond this point :-) ; ;sanity checks ; # 121 "/u/exp/stan/nmr/lists/pp/hncacbf.wfu" ; aqseq 321 ; define delay TAUA1 define delay TAUA2 define delay TAUB define delay TAUC define delay TAUC1 define delay TAUF1 define delay TAUF2 define delay TAUW define delay TN define delay C_H_N define pulse H1_90 define pulse H1_180 define pulse H1_S90 define pulse N15_90 define pulse N15_180 define pulse CAB_90 define pulse CAB_180 "d11=100m" ;disk i/o "d12=10u" ;power switching etc. "d13=5u" ;a short delay "d14=60u" ;ip,id etc. "d16=300u" ;gradient recovery "d17=50u" ;short gradient recovery "H1_90=p1" "H1_180=p1*2" "H1_S90=p10" "N15_90=p2" "N15_180=p2*2" "CAB_90=p3" "CAB_180=p3*2" "TAUW=d21" "TAUA1=2.25m-p15-d16-d13" "TAUA2=2.25m-TAUW*2.5-H1_90*2.385-p17-d16-d13" # 161 "/u/exp/stan/nmr/lists/pp/hncacbf.wfu" "TAUB=5.4m" "TN=12.0m" "TAUC=TN-CAB_180" "TAUC1=TAUC+CAB_180-TAUB-p11-d13-d12" # 170 "/u/exp/stan/nmr/lists/pp/hncacbf.wfu" "TAUF1=d3-p20-d17-p11-d12*3-d13*7" "TAUF2=TAUF1-d13" "d10=TN-d13*2-d12" "d30=TN-TAUB-CAB_180-p11-d12*2-d13*5" "d0=((cnst0*2+1)*in0-CAB_90*1.273-N15_180-d12*2-d13*4)/2" ; "d0=3u" "d22=(TAUW/2)-N15_90" "C_H_N=N15_90-H1_90" "d31=2*TAUA1+2*TAUB+TAUC1+TAUC+2*TAUF1+2*TAUF2+2*TAUA2+4*TAUW" # 195 "/u/exp/stan/nmr/lists/pp/hncacbf.wfu" # 1 "/u/exp/stan/nmr/lists/pp//Avance.incl" ;Avance.incl ; ;version 99/08/02 # 204 "/u/exp/stan/nmr/lists/pp/hncacbf.wfu" # 1 "/u/exp/stan/nmr/lists/pp//Gradfa.incl" ;Gradfa.incl - include file for Gradient Spectroscopy ;avance-version ; $Id: pulseprogram,v 1.1 2021/02/10 21:52:55 bmrbsvc Exp $ ; define list EA= # 205 "/u/exp/stan/nmr/lists/pp/hncacbf.wfu" 1 ze 2 d11 do:f2 d14 3 d14 d14 d14 d14 4 d14 d14 d14 5 d14 d14 d14 d14 6 d14 setnmr8^4 d14 setnmr2^0 # 1 "/u/exp/stan/nmr/lists/pp//faexptcorr.incl" ; This is here to trick "expt" to calculate the experiment time ; correctly. if "1 == 1" goto nod31 d31 nod31, d13 # 224 "/u/exp/stan/nmr/lists/pp/hncacbf.wfu" d1 pl1:f1 pl2:f2 d14 setnmr2|0 d14 setnmr8|4 (N15_90 ph0):f2 d13 p17:ngrad:c34 ;400u, 10G/cm d16 (H1_90 ph0):f1 d13 p15:ngrad:c34 ;500u, 4G/cm d16 TAUA1 (C_H_N H1_180 ph0):f1 (N15_180 ph0):f2 TAUA1 d13 p15:ngrad:c34 ;500u, 4G/cm d16 (H1_90 ph1):f1 d13 # 247 "/u/exp/stan/nmr/lists/pp/hncacbf.wfu" d12 pl10:f1 (H1_S90 ph21:r):f1 ;2ms 90 H1 pulse at phase x d13 p16:ngrad:c34 ;1.0m, 10G/cm d16 d13 \n d12 pl0:f3 \n d13 cpds3:f3 (N15_90 ph11):f2 TAUB d13 \n d12 pl11:f1 \n p11:f1 ph1 \n d13 cpds1:f1 TAUC1 d13 do:f3 \n d12 pl3:f3 \n d13 (N15_180 ph0):f2 d13 (CAB_180 ph0):f3 d13 \n d12 pl0:f3 \n d13 cpds3:f3 TAUC (N15_90 ph0):f2 d13 do:f1 \n p11:f1 ph3 \n d13 \n d12 pl1:f1 d13 do:f3 \n d12 pl3:f3 \n d13 d13 p18:ngrad:c34 ;1.0m, -5G/cm d16 d13 d13 \n d12 pl11:f1 \n p11:f1 ph1 \n d13 cpds1:f1 (CAB_90 ph22):f3 d13 \n d12 pl0:f3 \n d13 cpds3:f3 TAUF1 d13 d13 do:f1 \n p11:f1 ph3 \n d13 \n d12 pl1:f1 d13 do:f3 \n d12 pl3:f3 \n d13 p20:ngrad:c34 ;100u, 20G/cm d17 (CAB_180 ph0):f3 d13 p20:ngrad:c34 ;100u, 20G/cm d17 d13 \n d12 pl0:f3 \n d13 cpds3:f3 d13 \n d12 pl11:f1 \n p11:f1 ph1 \n d13 cpds1:f1 d13 TAUF2 d13 do:f3 \n d12 pl3:f3 \n d13 (CAB_90 ph1):f3 d13 \n d12 pl0:f3 \n d13 cpds3:f3 d0 (N15_180 ph0):f2 d0 d13 do:f3 \n d12 pl3:f3 \n d13 # 299 "/u/exp/stan/nmr/lists/pp/hncacbf.wfu" (CAB_90 ph23):f3 d13 \n d12 pl0:f3 \n d13 cpds3:f3 TAUF1 d13 d13 do:f1 \n p11:f1 ph3 \n d13 \n d12 pl1:f1 d13 do:f3 \n d12 pl3:f3 \n d13 p20:ngrad:c34 ;100u, 30G/cm d17 (CAB_180 ph0):f3 d13 p20:ngrad:c34 ;100u, 30G/cm d17 d13 \n d12 pl0:f3 \n d13 cpds3:f3 d13 \n d12 pl11:f1 \n p11:f1 ph1 \n d13 cpds1:f1 d13 TAUF2 d13 do:f3 \n d12 pl3:f3 \n d13 (CAB_90 ph24):f3 d13 d13 do:f1 \n p11:f1 ph3 \n d13 \n d12 pl1:f1 p19:ngrad:c34 ;600u, 10G/cm d16 d13 \n d12 pl0:f3 \n d13 cpds3:f3 d13 \n d12 pl11:f1 \n p11:f1 ph1 \n d13 cpds1:f1 (N15_90 ph13):f2 d10 d13 do:f3 \n d12 pl3:f3 \n d13 (N15_180 ph14):f2 d13 (CAB_180 ph0):f3 d13 \n d12 pl0:f3 \n d13 cpds3:f3 d30 d13 do:f1 \n p11:f1 ph3 \n d13 \n d12 pl1:f1 TAUB (N15_90 ph0):f2 d13 do:f3 \n d12 pl3:f3 \n d13 d13 p15:ngrad:c34 ;500u, 8G/cm # 341 "/u/exp/stan/nmr/lists/pp/hncacbf.wfu" d16 pl10:f1 (H1_S90 ph21:r):f1 ;2ms 90 H1 pulse at phase x d13 d12 pl1:f1 (H1_90 ph2):f1 d13 TAUA2 pl1:f1 p17:ngrad:c34 ;700u, 40G/cm d16 (H1_90*0.231 ph1):f1 TAUW (H1_90*0.692 ph1):f1 TAUW (H1_90*1.462 ph1):f1 d22 (N15_180 ph0):f2 d22 (H1_90*1.462 ph3):f1 TAUW (H1_90*0.692 ph3):f1 TAUW (H1_90*0.231 ph3):f1 d13 p17:ngrad:c34 ;700u, 40G/cm d16 pl12:f2 TAUA2 go=2 ph31 cpds2:f2 # 373 "/u/exp/stan/nmr/lists/pp/hncacbf.wfu" d11 do:f2 wr #0 if #0 zd d14 ip13 lo to 3 times 2 d14 dd10 d14 id30 d14 ip31 d14 ip31 lo to 4 times l4 d14 rd10 d14 rd30 # 388 "/u/exp/stan/nmr/lists/pp/hncacbf.wfu" d14 ip23 d14 ip24 lo to 5 times 2 d14 id0 d14 ip31 d14 ip31 lo to 6 times l6 d14 setnmr8^4 d14 setnmr2^0 exit ph0=0 ph1=1 ph2=2 ph3=3 ph11=0 2 ph13=0 ph14=0 0 0 0 2 2 2 2 ph22=0 0 2 2 ph23=1 1 1 1 3 3 3 3 ph24=0 0 0 0 0 0 0 0 2 2 2 2 2 2 2 2 ph21=(360) 0 ; phase x, adjust for any phase diff. between pl1 and sp1/pl110 ph31=0 2 2 0 0 2 2 0 2 0 0 2 2 0 0 2