KEY RESOURCES TABLE.

REAGENT or RESOURCE	SOURCE	IDENTIFIER
Bacterial and Virus Strains
STBL3	ThermoFisher	C737303
T7 Express lysY/Iq Competent E. coli (High Efficiency)	NEB	C3013
Chemicals, Peptides, and Recombinant Proteins
FBS, USA, Seradigm Premium	VWR	97068-085
KOD Hot Start DNA Polymerase	Millipore Sigma	71086-3
Proteinase K	NEB	P8107S
Tn5	F. Zhang Lab	-
Qiaprep spin miniprep kit	Qiagen	27106
IPTG	Millipore Sigma	I6758
cOmplete protease inhibitor	Millipore Sigma	11697498001
Benzonase	Millipore Sigma	E1014-25KU
Chitin resin	NEB	S6651L
OptiMEM	ThermoFisher	31985070
E-Gel™ EX Agarose Gels, 2%	ThermoFisher	G402002
GeneJuice	Millipore Sigma	70967-3
SF Cell Line 4D-Nucleofector® X Kit	Lonza	V4XC-2012
SE Cell Line 4D-Nucleofector® X Kit	Lonza	V4XC-1012
Puromycin	ThermoFisher	A1113802
NextSeq 500/550 High Output Kit v2, 75 cycles	Illumina	FC-404-2005
NextSeq 500/550 High Output Kit v2, 150 cycles	Illumina	FC-404-2002
Nuclease-Free Duplex Buffer	IDT	11-01-03-01
Deposited Data
Deep Sequencing data	SRA	PRJNA602092
Experimental Models: Cell Lines
HEK 293T	Gift from Veit Hornung	-
U-2 OS	ATCC	HTB-96
K562	Millipore Sigma	89121407-1VL
Oligonucleotides
/5Phos/CTGTCTCTTATACA/3ddC/	IDT	Transposon ME
GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG	IDT	Transposon read 2
/5phos/G*T*TGTGAGCAAGGGCGAGGAGGATAACGCCTCTCTCCCAGCGACT*A*T	IDT	TTISS donor sense
/5phos/A*T*AGTCGCTGGGAGAGAGGCGTTATCCTCCTCGCCCTTGCTCACA*A*C	IDT	TTISS donor antisense
GTCGCTGGGAGAGAGGCGTTATC	IDT	TTISS PCR fwd 1
AATGATACGGCGACCACCGAGATCTACACTATAGCCTACACTCTTTCCCTACACGACGCTCTTCCGATCTTTATCCTCCTCGCCCTTGCTCAC	IDT	TTISS PCR fwd 2
CAAGCAGAAGACGGCATACGAGATCGAGTAATGTCTCGTGGGCTCGGAGATGTGT	IDT	TTISS PCR rev BC1
CAAGCAGAAGACGGCATACGAGATTCTCCGGAGTCTCGTGGGCTCGGAGATGTGT	IDT	TTISS PCR rev BC2
CAAGCAGAAGACGGCATACGAGATAATGAGCGGTCTCGTGGGCTCGGAGATGTGT	IDT	TTISS PCR rev BC3
CAAGCAGAAGACGGCATACGAGATGGAATCTCGTCTCGTGGGCTCGGAGATGTGT	IDT	TTISS PCR rev BC4
CAAGCAGAAGACGGCATACGAGATTTCTGAATGTCTCGTGGGCTCGGAGATGTGT	IDT	TTISS PCR rev BC5
CAAGCAGAAGACGGCATACGAGATACGAATTCGTCTCGTGGGCTCGGAGATGTGT	IDT	TTISS PCR rev BC6
CAAGCAGAAGACGGCATACGAGATAGCTTCAGGTCTCGTGGGCTCGGAGATGTGT	IDT	TTISS PCR rev BC7
CAAGCAGAAGACGGCATACGAGATGCGCATTAGTCTCGTGGGCTCGGAGATGTGT	IDT	TTISS PCR rev BC8
CAAGCAGAAGACGGCATACGAGATCATAGCCGGTCTCGTGGGCTCGGAGATGTGT	IDT	TTISS PCR rev BC9
CAAGCAGAAGACGGCATACGAGATTTCGCGGAGTCTCGTGGGCTCGGAGATGTGT	IDT	TTISS PCR rev BC10
CAAGCAGAAGACGGCATACGAGATGCGCGAGAGTCTCGTGGGCTCGGAGATGTGT	IDT	TTISS PCR rev BC11
CAAGCAGAAGACGGCATACGAGATCTATCGCTGTCTCGTGGGCTCGGAGATGTGT	IDT	TTISS PCR rev BC12
CAAGCAGAAGACGGCATACGAGATTGTAGTGCGTCTCGTGGGCTCGGAGATGTGT	IDT	TTISS PCR rev BC13
CAAGCAGAAGACGGCATACGAGATGCGTCGACGTCTCGTGGGCTCGGAGATGTGT	IDT	TTISS PCR rev BC14
CAAGCAGAAGACGGCATACGAGATGGTCTTCTGTCTCGTGGGCTCGGAGATGTGT	IDT	TTISS PCR rev BC15
CAAGCAGAAGACGGCATACGAGATAAATGTCCGTCTCGTGGGCTCGGAGATGTGT	IDT	TTISS PCR rev BC16
CAAGCAGAAGACGGCATACGAGATGTTGAAACGTCTCGTGGGCTCGGAGATGTGT	IDT	TTISS PCR rev BC17
CAAGCAGAAGACGGCATACGAGATTCTTTACGGTCTCGTGGGCTCGGAGATGTGT	IDT	TTISS PCR rev BC18
CAAGCAGAAGACGGCATACGAGATATGCCTGGGTCTCGTGGGCTCGGAGATGTGT	IDT	TTISS PCR rev BC19
CAAGCAGAAGACGGCATACGAGATCAATAAGGGTCTCGTGGGCTCGGAGATGTGT	IDT	TTISS PCR rev BC20
CAAGCAGAAGACGGCATACGAGATCGCCGTAAGTCTCGTGGGCTCGGAGATGTGT	IDT	TTISS PCR rev BC21
CAAGCAGAAGACGGCATACGAGATTAAGGCTTGTCTCGTGGGCTCGGAGATGTGT	IDT	TTISS PCR rev BC22
CAAGCAGAAGACGGCATACGAGATTTGCTGCCGTCTCGTGGGCTCGGAGATGTGT	IDT	TTISS PCR rev BC23
CAAGCAGAAGACGGCATACGAGATCTCAATGTGTCTCGTGGGCTCGGAGATGTGT	IDT	TTISS PCR rev BC24
AATGATACGGCGACCACCGAGATCTACACTATAGCCTACACTCTTTCCCTACACGACGctcttccgatctCTTATCGTCGTCATCCTTGT	IDT	TTISS PCR prime +24 fwd a
AATGATACGGCGACCACCGAGATCTACACTATAGCCTACACTCTTTCCCTACACGACGctcttccgatctGATTACAAGGATGACGACGA	IDT	TTISS PCR prime +24 fwd b
GGCTTGTCGACGACGGCGGTC	IDT	TTISS PCR prime +38 fwd a1
AATGATACGGCGACCACCGAGATCTACACTATAGCCTACACTCTTTCCCTACACGACGctcttccgatctGACGGCGGTCTCCGTCGTCAG	IDT	TTISS PCR prime +38 fwd a2
ATGATCCTGACGACGGAGACCG	IDT	TTISS PCR prime +38 fwd b1
AATGATACGGCGACCACCGAGATCTACACTATAGCCTACACTCTTTCCCTACACGACGctcttccgatctGACGGAGACCGCCGTCGTCGA	IDT	TTISS PCR prime +38 fwd b2
Recombinant DNA
pTBX1-Tn5	Addgene	#60240
pX165	Addgene	#48137
pCMV-PE2	Addgene	#132775
pU6-pegRNA-GG-acceptor	Addgene	#132777
pX165-Sniper-Cas9	This study	-
pX165-LZ3 Cas9	This study	-
pX165-HiFi Cas9	This study	-
pX165-eSpCas9	This study	-
pX165-Cas9-HF1	This study	-
pX165-HypaCas9	This study	-
pX165-xCas9	This study	-
pX165-evoCas9	This study	-
Software and Algorithms
BrowserGenome	BrowserGenome.org	-
Elevation scoring	crispr.ml	-
GuideScan	guidescan.com	-
FORECasT	partslab.sanger.ac.uk/FORECasT	-
inDelphi	indelphi.giffordlab.mit.edu/single	-
Lindel	github.com/shendurelab/Lindel	-
Other
Bench Protocol	(this paper)	Methods S1