# 1 "/u/exp/stan/nmr/lists/pp/hncacbf.s" ;hncacbf.fa ;3D HNCACB, gradient enhanced with Watergate reverse INEPT (3-9-19), ; minimal H2O saturation and optional 2H decoupling. ; Modified for CryoProbe use. ;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 2021/02/11 01:09:12 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, f5: 2H (channel assignments may be changed below) ;Gradient program: fahncacbf ;o1p: 4.7ppm, ;o2p: 118ppm, ;o3p: 43ppm, ;o5p:: 4.5ppm ; ;d1: relaxation delay=d1+0.1003s ;p1 90 H1 at pl1 ;p11 90 2.0 ms H1 semi-selective H2O flip-back pulse at power: ; 1) pl21 (zgsel1.fa), with phcor21; ; 2) pl22 (zgsel2.fa), with phcor22. ; Check for a possible phase difference between hard H1 and soft H1 ;p10 90 H at pl10 (H1 composite decoupling) ;p2 90 N at pl2 ;p3 90 Cab at pl3 (for 90 Cab), typ. 17kHz at 500MHz ;p25 90 2H pulse at pl15 ;pl0 120dB ;d21: 1/(2*dNu), dNu=Nu(NH)-Nu(H2O) ; ;H1 Waltz-16x or DIPSI-2x (cpdprg1), using p90 (PCPD) at pl10 ;N15 Waltz-16 (cpdprg2), using p90 (PCPD) at pl12 ;C' Compensated SEDUCE1 decoupling of C', using p90 (PCPD) at sp3, ; shape seduce1c5/6, offset 0, cpdprg3 waltz16sp3. Select a pulse ; length (PCPD) that gives the right offset and adjust sp3 to make ; PCPD a 90 dgr pulse. ;H2 Waltz-16 (cpdprg5), using p90 (PCPD5) at pl15 ;N15 evolution: ; in10=in30, SW(N)=1/2*in10 ; l4 complex points; max. is (d10/in10)+1 ; Process as States (although data is acquired as States-TPPI) ; N15 chemical shift axis is reversed: set reverse to true. ;C13 evolution: ; in0, SW(C)=1/2*in0 ; l6 complex points ; set cnst0 to 0 (preferably) or 1 to make d0 the smallest possible ; positive delay. cnst0=0 gives (90,-180) phase distortion in F1. ; cnst0=1 gives (270,-540) phase distortion in F1 (use LP to correct). ;ns=8, 16, ..., ds=8, 16,... ; ;Recommendations for triple-axis (single-axis) gradients: ;gpz1: 12% (12%) ;gpz2: 6% (6%) ;gpz3: 15% (15%) ;gpz4: -7% (-7%) ;gpz7: 30% (30%) ;gpz8: 44% (44%) ;gpz9: 15% (15%) ;gpz12: 10% (10%) ;gpx13: 54% adjust for magic angle (0%) ;gpz13: 30% (55%) ;gpnam1: sine.50 ;gpnam2: sine.50 ;gpnam3: sine.100 ;gpnam4: sine.100 ;gpnam7: sine.10 ;gpnam8: sine.10 ;gpnam9: sine.50 ;gpnam12: sine.50 ;gpnam13: sine.100 ; ;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 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 ; # 108 "/u/exp/stan/nmr/lists/pp/hncacbf.s" ;#ifndef ; comment out for 2D w/o C13 evolution ;#undef ; uncomment for both Ca and Cb peaks, otherwise Cb only ;#endif ; aqseq 321 ; define delay TAUA define delay TAUA2 define delay TAUA13 define delay TAUB define delay TAUB1 define delay TAUC define delay TAUC1 define delay TAUC2 define delay TAUF define delay TAUF1 define delay TAUF2 define delay TAUF3 define delay TAUF4 define delay TAUW define delay TAUW1 define delay TN define delay CEN_HN1 define pulse GRAD1 define pulse GRAD2 define pulse GRAD3 define pulse GRAD4 define pulse GRAD5 define pulse GRAD6 define pulse GRAD7 define pulse GRAD8 define pulse GRAD9 define pulse GRAD10 define pulse GRAD11 define pulse GRAD12 define pulse GRAD13 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=H1_90*2" "H1_S90=p10" "N15_90=p2" "N15_180=N15_90*2" "CAB_90=p3" "CAB_180=CAB_90*2" "GRAD1=600u" "GRAD2=500u" "GRAD3=1.0m" "GRAD4=1.0m" "GRAD7=100u" "GRAD8=100u" "GRAD9=600u" "GRAD12=500u" "GRAD13=700u" "TAUW=d21" "TAUW1=(TAUW/2)-N15_90" "TAUA=2.3m" # 183 "/u/exp/stan/nmr/lists/pp/hncacbf.s" "TAUB=5.4m" "TN=12.0m" "TAUC=12.0m" "TAUF=3.5m" ; 3.5 ms for Ca and Cb # 192 "/u/exp/stan/nmr/lists/pp/hncacbf.s" "TAUA2=TAUA-GRAD2-d16-d13" "TAUA13=TAUA-TAUW*2.5-H1_90*2.385-GRAD13-d16-d13" "TAUB1=TAUB-d12-d13*2" "TAUC1=TAUC-CAB_180-d12*2-d13*5" "TAUC2=TAUC-TAUB-p10-d12*2-d13*4" # 203 "/u/exp/stan/nmr/lists/pp/hncacbf.s" "TAUF1=TAUF-GRAD7-d17-p10-d12*3-d13*7" "TAUF2=TAUF-GRAD7-d17-p10-d12*3-d13*8" "TAUF3=TAUF-GRAD8-d17-p10-d12*3-d13*7" "TAUF4=TAUF-GRAD8-d17-p10-d12*3-d13*8" "d10=TN-d13*4-d12*2" "d30=TN-TAUB-CAB_180-p10-d12*2-d13*5" "d0=((cnst0*2+1)*in0-CAB_90*1.273-N15_180-d12*4-d13*8)/2" "CEN_HN1=N15_90-H1_90" # 224 "/u/exp/stan/nmr/lists/pp/hncacbf.s" "d31=2*(GRAD2+TAUA2+TAUB1+GRAD7+GRAD8+TAUA13+GRAD13+2*TAUW+TAUW1)+GRAD1+GRAD3+TAUC1+TAUC2+GRAD4+TAUF1+TAUF2+TAUF3+TAUF4+GRAD9+GRAD12" # 1 "/u/exp/stan/nmr/lists/pp//Avance.incl" ;Avance.incl ; ;version 99/08/02 # 237 "/u/exp/stan/nmr/lists/pp/hncacbf.s" # 1 "/u/exp/stan/nmr/lists/pp//Grad.incl" ;Grad.incl - include file for Gradient Spectroscopy ;avance-version ;version 99/02/19 define list EA= # 238 "/u/exp/stan/nmr/lists/pp/hncacbf.s" 1 ze 2 d13 do:f2 d13 setnmr8^4 d11 setnmr2^0 d14 3 d14 d14 d14 d14 4 d14 d14 d14 5 d14 d14 d14 d14 6 d13 # 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 # 258 "/u/exp/stan/nmr/lists/pp/hncacbf.s" d1 pl1:f1 pl2:f2 d13 setnmr2|0 d13 setnmr0|34 d13 setnmr8|4 (N15_90 ph0):f2 d13 GRAD1:gp1 ;600u, 8G/cm, sine.50 d16 (H1_90 ph0):f1 d13 GRAD2:gp2 ;500u, 4G/cm, sine.50 d16 TAUA2 (CEN_HN1 H1_180 ph0):f1 (N15_180 ph0):f2 TAUA2 d13 GRAD2:gp2 ;500u, 4G/cm, sine.50 d16 (H1_90 ph1):f1 (d13 d12 pl22 H1_S90 ph22:r d13 d12 pl1):f1 ;2ms 90 H1 pulse at phase x d13 GRAD3:gp3 ;1.0m, 10G/cm, sine.100 d16 (N15_90 ph11):f2 d13 \n d12 pl0:f3 \n d13 cpds3:f3 TAUB1 d13 \n d12 pl10:f1 \n p10:f1 ph1 \n d13 cpds1:f1 TAUC2 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 TAUC1 d13 do:f3 \n d12 pl3:f3 \n d13 (N15_90 ph0):f2 d13 do:f1 \n p10:f1 ph3 \n d13 \n d12 pl1:f1 d13 GRAD4:gp4 ;1.0m, -5G/cm, sine.100 d16 d13 d13 \n d12 pl10:f1 \n p10:f1 ph1 \n d13 cpds1:f1 (CAB_90 ph15):f3 d13 \n d12 pl0:f3 \n d13 cpds3:f3 TAUF1 d13 d13 do:f1 \n p10:f1 ph3 \n d13 \n d12 pl1:f1 d13 do:f3 \n d12 pl3:f3 \n d13 GRAD7:gp7 ;100u, 20G/cm, sine.10 d17 (CAB_180 ph0):f3 d13 GRAD7:gp7 ;100u, 20G/cm, sine.10 d17 d13 \n d12 pl0:f3 \n d13 cpds3:f3 d13 \n d12 pl10:f1 \n p10: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 d13 do:f3 \n d12 pl3:f3 \n d13 (N15_180 ph0):f2 d13 \n d12 pl0:f3 \n d13 cpds3:f3 d0 d13 do:f3 \n d12 pl3:f3 \n d13 # 329 "/u/exp/stan/nmr/lists/pp/hncacbf.s" (CAB_90 ph16):f3 d13 \n d12 pl0:f3 \n d13 cpds3:f3 TAUF3 d13 d13 do:f1 \n p10:f1 ph3 \n d13 \n d12 pl1:f1 d13 do:f3 \n d12 pl3:f3 \n d13 GRAD8:gp8 ;100u, 30G/cm, sine.10 d17 (CAB_180 ph0):f3 d13 GRAD8:gp8 ;100u, 30G/cm, sine.10 d17 d13 \n d12 pl0:f3 \n d13 cpds3:f3 d13 \n d12 pl10:f1 \n p10:f1 ph1 \n d13 cpds1:f1 d13 TAUF4 d13 do:f3 \n d12 pl3:f3 \n d13 (CAB_90 ph17):f3 d13 d13 do:f1 \n p10:f1 ph3 \n d13 \n d12 pl1:f1 GRAD9:gp9 ;600u, 10G/cm, sine.50 d16 d13 \n d12 pl10:f1 \n p10:f1 ph1 \n d13 cpds1:f1 (N15_90 ph13):f2 d13 \n d12 pl0:f3 \n d13 cpds3:f3 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 p10:f1 ph3 \n d13 \n d12 pl1:f1 TAUB1 d13 do:f3 \n d12 pl3:f3 \n d13 (N15_90 ph0):f2 d13 GRAD12:gp12 ;500u, 7G/cm, sine.50 d16 (d13 d12 pl21 H1_S90 ph21:r d13 d12 pl1):f1 ;2ms 90 H1 pulse at phase x (H1_90 ph2):f1 d13 GRAD13:gp13 ;700u, 40G/cm, sine.100 d16 TAUA13 (H1_90*0.231 ph1):f1 TAUW (H1_90*0.692 ph1):f1 TAUW (H1_90*1.462 ph1):f1 TAUW1 (N15_180 ph0):f2 TAUW1 (H1_90*1.462 ph3):f1 TAUW (H1_90*0.692 ph3):f1 TAUW (H1_90*0.231 ph3):f1 TAUA13 GRAD13:gp13 ;700u, 40G/cm, sine.100 d16 pl12:f2 d13 setnmr0^34 go=2 ph31 cpds2:f2 # 396 "/u/exp/stan/nmr/lists/pp/hncacbf.s" d11 do:f2 wr #0 if #0 zd d13 setnmr8^4 d13 setnmr2^0 d14 ip13 lo to 3 times 2 d14 dd10 d14 id30 d14 ip31 d14 ip31 lo to 4 times l4 d14 rd10 d14 rd30 # 413 "/u/exp/stan/nmr/lists/pp/hncacbf.s" d14 ip16 d14 ip17 lo to 5 times 2 d14 id0 d14 ip31 d14 ip31 lo to 6 times l6 exit ph0=0 ph1=1 ph2=2 ph3=3 ph11=0 2 ph13=0 ph14=0 0 0 0 2 2 2 2 ph15=0 0 2 2 ph16=1 1 1 1 3 3 3 3 ph17=0 0 0 0 0 0 0 0 2 2 2 2 2 2 2 2 ph21=0 ph22=0 ph31=0 2 2 0 0 2 2 0 2 0 0 2 2 0 0 2