;Mrna_h5c5c4n3n1
;tested by anda 28.12.2011
;edited MS (28.10.2011)
;Erna_h5c5c4n3n1.t4
;rna_h5c5c4n3n1_etgpsi.t4.
;avance-version (03/02/28)
; 
; WARNING: consider that Bruker 13C calibration (TSP) is 2.7 ppm smaller than IUPAC (DSS)
; ==> use 2.7ppm smaller 13C offsets compared to BMRB reference chemical shifts

#include <Avance.incl>
#include <Grad.incl>
#include <Delay.incl>
 
"p2=p1*2"
"p22=p21*2"
"p8=500u"
"p16=1m"

"in0=inf1/4"
"l0=td1/2"

"cnst21=165.77"
"cnst22=98.67"

"d1=1s"
"d11=30m"
"d16=200u"
"d4=1.4m"
"d22=4.1m"
"d23=19.0m" 	; 19-21ms
"d25=25.0m"	; ~1/4J(N3,N1)
 
"d0=3u"

"DELTA=p2*2+p8+d0*4-4u"
"DELTA1=p16+d16+8u"
"DELTA2=d4-4u"
"DELTA3=d22-p16-d16"
"DELTA4=d25/2-p16-d16-p29/2-p21*2/3.1416"
"DELTA5=d25-p16*2-d16*2-p29"
 
"spoff2=0"
"spoff3=0"
"spoff5=bf2*((cnst21-cnst22)/1000000)"
;"spoff8=0"
;"spoff9=0"
"spoff13=0"
"spoff19=0"
 
1 ze
2 50m do:f2 
  d11 pl1:f1 pl12:f2
3 10m
4 d1
  30u fq=cnst23(bf ppm):f3			; CHECK if oK!!!
  30u fq=cnst22(bf ppm):f2
 
  (p1 ph1)
  DELTA2 pl0:f2
  4u
  (center (p2 ph1) (p14:sp3 ph1):f2 )
  4u
  DELTA2 UNBLKGRAD
;  p28 ph1
;  4u
  (p1 ph2)
 
  (p13:sp2 ph3):f2
  d22
  (p14:sp5 ph1):f2
  4u
  (p14:sp3 ph1):f2
  d22
  (p14:sp5 ph1):f2
  4u
  (p13:sp8 ph2):f2
 
  4u
  30u fq=cnst21(bf ppm):f2
 
  (p13:sp2 ph1):f2
  d23 pl0:f3
  (center (p14:sp3 ph1):f2 (p30:sp9 ph1):f3 )
  d23
  (p13:sp8 ph2):f2
 
  p16:gp3
  d16 pl3:f3
 
  (p21 ph8):f3
  DELTA4
  p16:gp4
  d16 pl0:f3
  (p29:sp19 ph10):f3
  p16:gp4
  d16
  DELTA5
  p16:gp4
  d16
  (p29:sp19 ph10):f3
  p16:gp4
  d16
  DELTA4 pl3:f3
  (p21 ph9):f3
 
  d0
  (p2 ph7)
  d0
  (p8:sp13 ph7):f2
  d0
  (p2 ph7)
  d0
  (p22 ph1):f3
  4u
  DELTA
 
  (p21 ph2):f3
  DELTA4
  p16:gp5
  d16 pl0:f3
  (p29:sp19 ph1):f3
  p16:gp5
  d16
  DELTA5
  p16:gp5
  d16
  (p29:sp19 ph1):f3
  p16:gp5
  d16
  DELTA4 pl3:f3
  (p21 ph1):f3
 
  (p13:sp2 ph2):f2
  d23 pl0:f3
  (center (p14:sp3 ph1):f2 (p30:sp9 ph1):f3 )
  d23
  (p13:sp8 ph1):f2
 
  4u
  30u fq=cnst22(bf ppm):f2
 
  (p13:sp2 ph2):f2
  4u
  (p14:sp5 ph1):f2
  d22
  (p14:sp3 ph1):f2
  4u
  (p14:sp5 ph1):f2
  DELTA3
  p16:gp1
  d16 pl1:f1 pl2:f2
 
  (center (p1 ph1) (p3 ph4):f2 )
  4u
  DELTA2 pl0:f2
  (center (p2 ph1) (p14:sp3 ph1):f2 )
  4u
  DELTA2 pl2:f2
  (center (p1 ph2) (p3 ph5):f2 )
  4u
  DELTA2 pl0:f2
  (center (p2 ph1) (p14:sp3 ph1):f2 )
  4u
  DELTA2
  (p1 ph1)
  DELTA1
  (p2 ph1)
  4u
  p16:gp2
  d16 pl12:f2
  4u BLKGRAD
  go=2 ph31 cpds2:f2
  30m do:f2 wr #0 if #0 zd ip8  
  10m ip9  
  10m ip10
  lo to 3 times 2
  10m id0
  lo to 4 times l0
exit
    
 
ph1=0
ph2=1
ph3=0 2
ph4=0 0 0 0 2 2 2 2
ph5=3 3 3 3 1 1 1 1
ph6=0
ph7=0 0 0 0 2 2 2 2
ph8=0 0 2 2
ph9=1
ph10=0
ph31=0 2 2 0 2 0 0 2
   
 
;o3p 180ppm 
;pl0 : 120dB
;pl1 : f1 channel - power level for pulse (default)
;pl2 : f2 channel - power level for pulse (default)
;pl3 : f3 channel - power level for pulse (default)
;pl12: f2 channel - power level for CPD/BB decoupling
;cnst21 165.8 C4 166-169)(cannot decouple Py C2 @154-159ppm)
;cnst22 98.7 (13C o2p on-resonance) decouple C5 (136-144ppm)
;cnst23 185 (15N o3p on-resonance) 
;sp2 p13 1.2m Q5.1000
;sp3 p14 660u Q3.1000
;sp5 p14 660u Q3.1000 (offset)
;sp8 p13 1.2m Q5tr.1000
;sp9 Q3.1000 460u (900 MHz) +/- 28ppm N3 @180ppm, 156-212ppm (156-162 Ura, 202-212 Cyt) decouple N4 (93-102 Cyt) 
;sp13 Crp60,0.5,20.1 invert all 13C during 15N evolution (+/- 13 kHz)
;sp19 2.6ms rna_phs8,0.7,4m.1 gB1max=3923 Hz (90 deg rect.= 63.7us)
;spnam2 Q5.1000
;spnam3 Q3.1000
;spnam5 Q3.1000 (offset)
;spnam8 Q5tr.1000
;spnam9 Q3.1000 460u 15N N3 @180ppm, 156-212ppm (156-162 Ura, 202-212 Cyt) decouple N4 (93-102 Cyt) 
;spnam13 Crp60,0.5,20.1 invert all 13C during 15N evolution (+/- 13 kHz)
;spnam19 2.6ms rna_phs8,0.7,4m.1 gB1max=3923 Hz (90 deg rect.= 63.7us)
;p1 f1 1H 90 degree high power pulse
;p2 f1 1H 180 degree high power pulse
;p3 f2 13C 90 degree high power pulse
;p8 f2 13C 500u Crp60,0.5,20.1
;p13  13C 1200u (+/- 10ppm) (900 MHz) Q5.1000 C5 (94-99ppm Cyt, 102-107 Ura; decouple C6 136-144) C4 (166-168 Cyt, 169 Ura)
;p14  13C 660u (+/- 9ppm) (900 MHz) Q3.1000 C5 (94-99ppm Cyt, 102-107 Ura) and  C4 (166-168 Cyt, 169 Ura)
;p16  500-1000u gradient pulse
;p22  f3 channel - 180 degree high power pulse
;p28  f1 channel - trim pulse
;p29 f3 15N 2.6ms (900 MHz), rna_phs8,0.7,4m.1 15N HS for refocussing
;p30 f3 15N 450u (900 MHz) Q3.1000 invert N3 (202-212 Cyt, 156-162 Ura) decouple N4 (93-120 Cyt)
;d0   incremented delay (2D)                         [3 usec]
;d1   relaxation delay; 1-5 * T1
;d4   1/(4J)XH
;d11  30m delay for disk I/O                             [30 msec]
;d16  200u gradient recovery
;d23 [19-21 msec]
;d25 25ms N3->N1 (2*d25=1/2J(N3,N1)               [25.0 msec]

;cnst2  = J(XH)
;in0  1/(2 * SW(X)) = DW(X)
;nd0  2
;NS: 1 * n
;DS: >= 16
;td1: number of experiments
;cpd2: decoupling according to sequence defined by cpdprg2
;pcpd2: f2 channel - 90 degree pulse for decoupling sequence
 
;for z-only gradients:
;gpz1: 80%
;gpz2: 20.1% for C-13, 8.1% for N-15
;gpz3: 30%
;gpz4: 11%
;gpz5: 7%
 
;use gradient files:
;gpnam1: SQSM10.100
;gpnam2: SQSM10.100
;gpnam3: SQSM10.100
;gpnam4: SQSM10.100
;gpnam5: SQSM10.100