;Leo Wong, lewo@nmr.mpibpc.mpg.de
;Sensitivity-enhanced HNcocaNH and HNcocancaNH (x-P-x) APSY


;p21   : 1000u
;p22   : 600u
;p30   : 800u
;p31   : 250u

;gpz0   : 15 %
;gpz1   : 25 %
;gpz2   : 30 %
;gpz3   : 35 %
;gpz4   : 40 %
;gpz5   : 45 %
;gpz6   : 50 %
;gpz7   : 55 %
;gpz8   : 60 %
;gpz9   : 65 %
;gpz10  : 70 %
;gpz11  : 75 %
;gpz12  : 80 %
;gpz30  : 80 %
;gpz31  : 25.91%

;d1    : relaxation delay

;cnst3: angle alpha
;cnst4: angle beta

;cnst8: 1H carrier in amide region (8.2 ppm)
;cnst9: Selective inversion offset (8.5 ppm)
;cnst20: Set to o1p
;cnst23: 15N carrier incl. proline (127 ppm)
;cnst25: 13C carrier for aliphatic (45 ppm)
;cnst26: Cb/g of proline (30 ppm)
;cnst27: Calpha of proline (64 ppm)
;cnst28: Chirp pulse offset (110 ppm)
;cnst29: CO chemical shift (offset, 176 ppm)
;cnst30: Ca chemical shift (offset, 56 ppm)

;;Shaped pulse length at 700 MHz;;
;p7    : 1300u
;p8    : 1300u
;p9    : 300u
;p11   : 330u
;p12   : 200u
;p14   : 760u
;p15   : 200u
;p19   : 240u
;p20   : 120u
;p24   : 400u
;p25   : 1300u
;p28   : 750u

;sp2   :  90 deg,  p7, Gauss_5%, H2O
;sp3   :  90 deg,  p7, Gauss_5%, H2O
;sp9   : 180 deg,  p9, IBurp2, Cali
;sp11  : 180 deg, p11, Reburp, Cali
;sp12  :  90 deg, p12, EBurp2, Calpha
;sp13  : 180 deg, p13, BLOCK.32, Calpha
;sp14  : 180 deg, p14, IBurp2, Calpha
;sp15  : 180 deg, p15, I3Snob, CO
;sp17  : 180 deg, p17, BLOCK.32, CO
;sp19  : 180 deg, p19, Q3, CO
;sp20  :  90 deg, p20, Gauss_5%, CO
;sp24  : 180 deg, p24, Crp60, Cali/aro
;sp25  : 180 deg, p25, I3Snob, HN
;sp28  : 180 deg, p28, Q3, Calpha Pro


#define GRADIENT1 10u p21:gp0 190u
#define GRADIENT3 10u p21:gp1 190u
#define GRADIENT4 10u p21:gp2 190u
#define GRADIENT5 10u p22:gp5 190u
#define GRADIENT6 10u p22:gp9 190u
#define GRADIENT7 10u p22:gp7 190u
#define GRADIENT8 10u p21:gp3 190u
#define GRADIENT9 10u p21:gp4 190u
#define GRADIENT11 10u p22:gp11 190u

;----------------------offsets

"p13=866000/(bf2*(cnst29-cnst30))"
"p17=866000/(bf2*(cnst29-cnst30))"

"spoff2=0.0"
"spoff3=bf1*((cnst20-cnst8)/1000000)"
"spoff9=bf2*((cnst25-cnst29)/1000000)"
"spoff11=bf2*((cnst25-cnst30)/1000000)"
"spoff12=0.0"
"spoff13=0.0"
"spoff14=0.0"
"spoff15=bf2*((cnst29-cnst30)/1000000)"
"spoff17=0.0"
"spoff19=0.0"
"spoff20=0.0"
"spoff23=0.0"
"spoff24=bf2*((cnst28-cnst30)/1000000)"
"spoff25=bf1*(cnst9/1000000)-o1"
"spoff28=bf2*((cnst27-cnst30)/1000000)"

"cnst14=abs(sin(0.017453293*cnst3))"
"cnst15=abs(cos(0.017453293*cnst3))"
"cnst16=abs(sin(0.017453293*cnst4))"
"cnst17=abs(cos(0.017453293*cnst4))"

"l1=td1/2"

"p2=2*p1"
"p6=2*p5"
"p35=(2/3)*p5"
"p36=(20/6)*p5"

;------------------------------basic delays
"d2 =2.7m"    ;1/2 H-N
"d4 =24.0m"   ;1/2 N-CA w/o CB-CA
"d5 =27.0m"   ;1/2 N-CA plus 2*CB-CA
"d7 =14.0m"   ;sqrt(2)*1/2 N-CA
"d8 =14.0m"   ;1/2 N-CO
"d9 =4.75m"   ;1/2 CO-CA

#ifdef Pro
"d58=d4-d9-p15-5u"
"d59=d4-p15-5u"
"d60=16.5m"    ;1/2 N-CA for proline plus 1/2 N-CO
"d62=d2-(0.63*p25)/4"
"d63=d60-d62-p26-5u-(p25-p24)/2"
#endif /*Pro*/
;----------------------fixed delays

define delay INEPT_N1
"INEPT_N1=d2-(p21+200u)-p5-p7-4u"
define delay INEPT_N2
"INEPT_N2=d2-(p21+200u)-p5"
define delay INEPT_N3
"INEPT_N3=d2-(p21+200u)-p5-10u-p8-2u"
define delay INEPT_N4
"INEPT_N4=d2-(p21+200u)-p5-10u"
define delay DELTA1
"DELTA1=p30+200u"
define delay DELTA2
"DELTA2=p31+200u"

;----------------------------indirect dimensions
"in1=inf1"
;----------------------H
"d11=d2-(p21+200u)-p6-5u"
"d20=5u"
"d21=d2-(p21+200u)"

"in11=cnst16*in1/2"
"in21=(d21-20u)/l1"
"in20=0u"
;----------------------N
"d12=10u"
"d16=d8-p17-10u"
"d22=10u"
"d23=d8"

"in12=cnst17*cnst14*in1/2"
"in23=(d23-20u)/l1"
"in22=0u"
;----------------------O
"d25=d9-0.63*p9"
;----------------------A
"d26=d5-d9-p15-5u-p11/2"
"d27=d5-p15-5u-p11/2"
"d57=d5-p11/2"
;----------------------M
"d30=d7"
"d29=10u"
"d6=d7-p13-10u-d2*2-p26-15u"
"d15=d7-p13-10u-DELTA1"
"d28=d7-p13-p15-20u"

"in15=cnst17*cnst15*in1/2"
"in30=(d30-20u)/l1"
"in6=in15"
"in29=0u"
;----------------------

#include <Avance.incl>

define loopcounter ST1CNT
#ifdef E125
  "ST1CNT = td1 / (8)"
#else
  "ST1CNT = td1 / (2)"
#endif

1  10u ze
;------------------------------------Switch constant-time for 15M
if "in30 < in15" goto 5
   10u
   "in30=in15"
   10u
goto 6
5  1m
   10u
   "in29=in15-in30"
   10u
6  1m
;------------------------------------Switch constant-time for 15N
if "in23 < in12" goto 7
   10u
   "in23=in12"
   10u
goto 8
7  1m
   10u
   "in22=in12-in23"
   10u
8  1m
;------------------------------------Switch constant-time for 1H
if "in21 < in11" goto 11
   10u
   "in21=in11"
   10u
goto 12
11 1m
   10u
   "in20=in11-in21"
   10u
12 1m
;------------------------------------
2  5m
   LBLSTS1, 5m
13 3m
14 3m
   LBLF1, 5m
   10u do:f3
   10u LOCKH_OFF
   d1
   10u fq=cnst29 (bf ppm):f2
   10u
   10u fq=cnst8 (bf ppm):f1
   10u
   10u pl2:f2
   10u pl3:f3
   10u LOCKH_ON
   10u
   (p8:sp3 ph3):f1
   2u
   2u pl1:f1
;-----------------------------------------first INEPT
   (p1 ph1):f1
   GRADIENT1
   d11
   (p6 ph20):f3
   d20
   (p2 ph20):f1
   GRADIENT1
   d21
;-----------------------------------------PEP
   (center (p1 ph11):f1 (p5 ph21):f3)
   2u
   (p8:sp3 ph11):f1
   GRADIENT3
   10u pl1:f1
   INEPT_N3
   (center (p2 ph21):f1 (p6 ph20):f3)
   GRADIENT3
   INEPT_N2
   (center (p1 ph20):f1 (p5 ph20):f3)
   2u
   (p8:sp3 ph22):f1
   GRADIENT4
   10u pl1:f1
   INEPT_N3
   (center (p2 ph20):f1 (p6 ph20):f3)
   GRADIENT4
   10u fq=0(sfo hz):f1
   INEPT_N4 pl11:f1
   (p5 ph23):f3
;-----------------------------------------end PEP
   (refalign (d12 p9:sp9 ph20 d16):f2 lalign (p26 ph21 5u cpds1 ph20):f1)
   (p17:sp17 ph20):f2        ;I3Snob?
   d22
   (p6 ph20):f3
   d23
   (p9:sp9 ph20):f2
   10u
   (p5 ph12):f3

   5u do:f1
   (p26 ph23):f1
;-----------------------------------------COzNz
   GRADIENT5
;-----------------------------------------
   (p20:sp20 ph2):f2
   (d25 p9:sp9 ph20 4u):f2
   (p19:sp19 ph23):f2
   (d25 p9:sp9 ph20 4u):f2
   (p20:sp20 ph13):f2
;-----------------------------------------CAzCOzNz
   GRADIENT6
   10u fq=cnst30 (bf ppm):f2
   10u
#ifdef Pro
   10u fq=cnst23 (bf ppm):f3
   10u
#endif /*Pro*/
;-----------------------------------------CAzCOzNz
   (p12:sp12 ph14):f2
   d9
   (p15:sp15 ph20):f2
   5u

#ifdef Pro
   d58
   (refalign (p28:sp28 ph20):f2 center (p5 ph21 3u p6 ph20 3u p5 ph21):f3)
   d59
#else
   d26
   (refalign (p11:sp11 ph20):f2 center (p6 ph20):f3)
   d27
#endif /*Pro*/

   (p15:sp15 ph20):f2
   1u
   4u pl2:f2
   (p3 ph21):f2
;-----------------------------------------NzCAz
   GRADIENT7
;-----------------------------------------For N(i)-N(i-2) relay over proline
#ifdef Pro
   (p26 ph21 5u cpds1 ph20):f1

   (p5 ph20):f3
   d63
   5u do:f1
   (p26 ph23):f1
   d62
   (refalign(p25:sp25 ph20):f1 center(p24:sp24 ph20):f2 center(p5 ph21 3u p6 ph20 3u p5 ph21):f3)
   d62 pl11:f1
   (p26 ph21 5u cpds1 ph20):f1
   d63
   (p5 ph21):f3
   4u
;-----------------------------------------CAzCOzNz
   (p12:sp12 ph20):f2
   d9
   (p15:sp15 ph20):f2
   5u
   d26
   (refalign (p11:sp11 ph20):f2 center (p5 ph21 3u p6 ph20 3u p5 ph21):f3)
   d27
   (p15:sp15 ph20):f2
   1u
   4u pl2:f2
   (p3 ph23):f2

   5u do:f1
   (p26 ph23):f1
;-----------------------------------------NzCAz
   GRADIENT11
   10u fq=0(sfo hz):f3
   10u
#endif /*Pro*/
;-----------------------------------------For N(i)-N(i-2) relay over non-proline
#ifdef nonPro
   (p26 ph21 5u cpds1 ph20):f1

   (p5 ph20):f3
   d4
   (refalign(p14:sp14 ph20):f2 center (p6 ph20):f3)
   d4 pl2:f2
   (p5 ph20):f3
   4u
;-----------------------------------------CAzNz
   (p3 ph20):f2
   d57
   (refalign (p11:sp11 ph20):f2 center (p6 ph20):f3)
   d57 pl2:f2
   (p3 ph20):f2

   5u do:f1
   (p26 ph23):f1
;-----------------------------------------NzCAz
   GRADIENT11
#endif /*nonPro*/
;-----------------------------------------NzCAz
   (p26 ph21 5u cpds1 ph20):f1

   (p5 ph20):f3
   d30
   (p6 ph20):f3
   d29
   (p13:sp13 ph20):f2

   (d6 5u do p26 ph23 10u pl1):f1 (d15 10u p30:gp30*plusminus 190u):f3 (d28 p15:sp15 ph20):f2
;-----------------------------------------PEP
   (center (p1 ph20):f1 (p5 ph20):f3)
   2u
   2u
   (p7:sp2 ph22):f1
   GRADIENT8
   INEPT_N1 pl1:f1
   (center (p2 ph20):f1 (p6 ph20):f3)
   GRADIENT8
   INEPT_N1
   (p7:sp2 ph23):f1
   2u
   2u pl1:f1
   (center (p1 ph23):f1 (p5 ph15):f3)
   GRADIENT9
   INEPT_N2
   (center (p2 ph20):f1 (p6 ph20):f3)
   GRADIENT9
   INEPT_N1
   (p7:sp2 ph22):f1
   2u
   2u pl1:f1
   (p1 ph20):f1
;-----------------------------------------end PEP
   DELTA2
   (p2 ph21):f1
   10u
   p31:gp31
   190u pl13:f3 LOCKH_OFF
;-----------------------------------------acquisition
   go=2 ph31 cpd3:f3

#ifdef E5
   1m do:f3 wr #0 if #0 zd ip15*2 igrad plusminus
#endif
#ifdef E125
   1m do:f3 wr #0 if #0 zd ip15*2 igrad plusminus
   lo to LBLSTS1 times 2
   3m ip12
   lo to 13 times 2
   3m ip11*2
   lo to 14 times 2
   goto 16
#endif
   lo to LBLSTS1 times 2
16 1m id11
   1m id20
   1m dd21
   1m id12
   1m id22
   1m dd23
   1m id15
   1m id29
   1m dd30
   1m id6
   lo to LBLF1 times ST1CNT
   10u do:f1
   10u do:f2
   10u do:f3
   10u LOCKH_OFF
exit

;-------------------phases for phasecycle
ph1= 0 2
ph2= 3 3 1 1
ph3= 2 0
ph8= 2
ph9= 3
ph31=0 2 2 0

;-------------------other phases
ph11=1
ph12=0
ph13=0
ph14=0
ph15=1

;-------------------fixed phases
ph20=0
ph21=1
ph22=2
ph23=3