Overview

In the process of constructing the Click Virus, we designed and utilized numerous structures. These structures primarily include Vaccinia virus A27L, Vaccinia virus A27L-3stop, Amber suppressor tRNA gene tandem, Azidophenylalanine-specific tyrosyl-tRNA synthetase, and other domains. We will briefly list all our parts on this page including "Basic Parts" and "Composite Parts".All the parts we use have been added to the registry.

New Basic Parts

	Type	Part number	Name	Brief introductions
1	Coding	BBa_K5142031	Vaccinia virus A27L	A membrane protein of vaccinia virus (VACV) which is important for forming extracellular enveloped virus (EEV).
2	Coding	BBa_K5142032	Vaccinia virus A27L-3stop	This part codes A27L-3stop, which is derived from the A27L mutation, where the codons of three phenylalanines at the N-terminus are changed to the amber suppressor codon UAG.
3	RNA	BBa_K5142033	Amber suppressor tRNA gene tandem	This part contains nine identical genes which can be transcribed into amber suppressor tRNA with an anticondon of CUA so that it can read the amber suppressor codon UAG
4	Coding	BBa_K5142034	Azidophenylalanine-specific tyrosyl-tRNA synthetase	This part includes the coding sequence (cds) of an azidophenylalanine-specific variant of E. coli tyrosyl-tRNA synthetase (pAzpaRS).
5	Coding	BBa_K5142035	Vaccinia virus A28L	This part includes the coding sequence (cds) of A28L, a membrane protein of vaccinia virus (VACV).
6	Regulatory	BBa_K5142036	Overlapping early/late poxviral promoter	This part can drive the transcription of its downstream cds with high efficiency ONLY in poxviral factory
7	Regulatory	BBa_K5142037	Reverse poxviral H5 promoter	This part includes a DNA fragment derived from the promoter of H5 gene of vaccinia virus. This part can drive the transcription of its downstream element with high efficiency ONLY in poxviral factory
8	Coding	BBa_K5142038	Reverse complementary GFP cds	This part contains the reverse complementary sequence of the coding sequence of green fluorescent protein (GFP).
9	Coding	BBa_K5142039	Vaccinia virus A26L (partial)	This part includes the promoter and partial coding sequence (cds) of A26L, a membrane protein of vaccinia virus (VACV).
10	DNA	BBa_K5142040	FOLR1-T2A-mCherry cds	This part contains the coding sequences (cds) of human folate receptor protein FOLR1 and mCherry which are in the same reading frame and separated by a T2A sequence.
11	DNA	BBa_K5142041	FOLR2-T2A-mCherry cds	This part contains the coding sequences (cds) of human folate receptor protein FOLR2 and mCherry which are in the same reading frame and separated by a T2A sequence.
12	Coding	BBa_K5142042	Reverse complementary GFP cds	This part contains the reverse complementary sequence of the coding sequence of green fluorescent protein (GFP).
13	Coding	BBa_K5142043	Vaccinia virus A26L (partial)	This part includes the promoter and partial coding sequence (cds) of A26L, a membrane protein of vaccinia virus (VACV).
14	Other	BBa_K5142048	Click Virus	This part is a genetically engineered vaccinia virus (VACV) with modified A27L protein containing three 4-azido-L-phenylalanine residues close to its N-terminus.

New Composite Part

	Type	Part number	Name	Brief introductions
1	DNA	BBa_K5142045	Compositive construct for A27L-3Stop mutagenesis	This part was used to replace the wild-type A27L coding sequence in vaccinia virus (VACV) genome with A27L-3Stop, via homologous recombination in VACV genome.

Basic Parts we used

	Part number	Name	Brief introductions	Group
1	BBa_K2217024	CMV Enhancer + CMV Promoter	It is composed of CMV Enhancer as well as CMV Promoter (BBa_K2217000 and BBa_K2217006 respectively) as constitutive promoters to enhance the transcription process.	iGEM17_AFCM-Egypt
2	BBa_J64997	T7 consensus -10 and rest	This part has been characterized for GFP expression.	Voigt Lab
3	BBa_K5142035	Vaccinia virus A28L	This part includes the coding sequence (cds) of A28L, a membrane protein of vaccinia virus (VACV).	HunanU
4	BBa_K5142038	Reverse complementary GFP cds	This part contains the reverse complementary sequence of the coding sequence of green fluorescent protein (GFP).
5	BBa_K5142037	Reverse poxviral H5 promoter	This part includes a DNA fragment derived from the promoter of H5 gene of vaccinia virus. This part can drive the transcription of its downstream element with high efficiency ONLY in poxviral factory	HunanU
6	BBa_K5142036	Overlapping early/late poxviral promoter	This part can drive the transcription of its downstream cds with high efficiency ONLY in poxviral factory	HunanU
7	BBa_K5142032	Vaccinia virus A27L-3stop	This part codes A27L-3stop, which is derived from the A27L mutation, where the codons of three phenylalanines at the N-terminus are changed to the amber suppressor codon UAG.	HunanU
8	BBa_K5142039	Vaccinia virus A26L (partial)	This part includes the promoter and partial coding sequence (cds) of A26L, a membrane protein of vaccinia virus (VACV).
9	BBa K3830009	f1 ori	f1 bacteriophage origin of replication; arrowindicates direction of(+)strand synthesis
10	BBa K1722006	SV40 promoter	SV40 enhancer and early promoter
11	BBa K4574000	SV40 ori	SV40 origin of replication
12	BBa K4574000	NeoR/KanR	aminoglycoside phosphotransferase fromTn5, confers resistance to neomycin, kanamycin, andG418
13	BBa K2543009	SV40 poly(A) signal	SV40 polyadenylation signal
14	BBa K5487110	M13 rev	common sequencing primer, one of multiple similarvariants
15	BBa K4673024	lac operator	The lac repressor binds to the lac operator to inhibittranscription in E, coli. This inhibition can berelieved by adding lactose orisopropyl-β-D-thiogalactopyranoside(IPTG).
16	BBa K1804001	lac promoter	promoter for the E. coli lac operon
17	BBa M50499	CAP binding site	CAP binding activates transcription in the presenceof cAMP.
18	BBa I50022	ori	high-copy-number colE1/pMB1/pBR322/pUc originof replication
19	BBa K1927002	AmpR	confers resistance to ampicillin, carbenicillin, andrelated antibiotics
20	BBa K3766184	AmpR promoter	promoter for the AmpR

	Part number	Name	Brief introductions
1	BBa K2287021	pRS-marker	pRS vectors,use to sequence yeast selectablemarker
2	BBa_K5142033	Amber suppressor tRNA gene tandem	This part contains nine identical genes which can be transcribed into amber suppressor tRNA with an anticondon of CUA so that it can read the amber suppressor codon UAG
3	BBa_K4278221	pBRrevBam	pBR322 vectors,tet region,downstream of BamHI,reverse primer
4	BBa_K2217024	CMV Enhancer + CMV Promoter	It is composed of CMV Enhancer as well as CMV Promoter (BBa_K2217000 and BBa_K2217006 respectively) as constitutive promoters to enhance the transcription process.
5	BBa K4818042	tet operator	bacterial operator 02 for the tetR and tetA genes
6	BBa_I712004	LNCX	Human CMV promoter, a constitutive expression promoter for use in mammalian cells.
7	BBa_K3200999	BGH-rev	Bovine growth hormone terminator, reverse primer.
8	BBa_K2591013	bGH poly(A) signal	bovine growth hormone polyadenylation signal
9	BBa K3830009	f1 ori	f1 bacteriophage origin of replication; arrowindicates direction of(+)strand synthesis
10	BBa K1722006	SV40 promoter	SV40 enhancer and early promoter
11	BBa K4574000	SV40 ori	SV40 origin of replication
12	BBa_K5142034	Azidophenylalanine-specific tyrosyl-tRNA synthetase	This part includes the coding sequence (cds) of an azidophenylalanine-specific variant of E. coli tyrosyl-tRNA synthetase (pAzpaRS).
13	BBa K1150016	HA	The human influenza hemagglutinin (HA)–tag enables the detection, isolation and purification of proteins.
14	BBa K2543009	SV40 poly(A) signal	SV40 polyadenylation signal
15	BBa K5487110	M13 rev	common sequencing primer, one of multiple similarvariants
16	BBa K4673024	lac operator	The lac repressor binds to the lac operator to inhibittranscription in E, coli. This inhibition can berelieved by adding lactose orisopropyl-β-D-thiogalactopyranoside(IPTG)
17	BBa K1804001	lac promoter	promoter for the E. coli lac operon
18	BBa M50499	CAP binding site	CAP binding activates transcription in the presenceof cAMP.
19	BBa_K4587001	L4440	L4440 vector
20	BBa I50022	ori	high-copy-number colE1/pMB1/pBR322/pUc originof replication
21	BBa K1927002	AmpR	confers resistance to ampicillin, carbenicillin, andrelated antibiotics
22	BBa K3766184	AmpR promoter	promoter for the AmpR

	Part number	Name	Brief introductions
1	BBa_K5487114	M13 fwd	common sequencing primer, one of multiple similar variants.
2	BBa_K2217024	CMV Enhancer + CMV Promoter	It is composed of CMV Enhancer as well as CMV Promoter (BBa_K2217000 and BBa_K2217006 respectively) as constitutive promoters to enhance the transcription process.
3	BBa_K4344028	Kozak sequence	The Kozak-sequence is the conserved consensus sequence next to the promoter, surrounding the AUG for the start of transcription in eukaryotes.
4	BBa_K5142040	FOLR1	the coding sequence(cds) of human folate receptor protein FOLR1
5	BBa_K1993019	T2A	T2A is a self-cleaving 2A peptide (The average length is 18–22 amino acids) that could be applied in expression of more than one gene in cells.
6	BBa_J06504	mCherry	the coding sequence (cds) of mCherry
7	BBa_K4818027	3xFLAG	The term "3xFLAG" refers to a protein tag used in molecular biology and biochemistry to facilitate the detection, purification, and characterization of proteins.
8	BBa K1722006	SV40 promoter	SV40 enhancer and early promoter
9	BBa K45740	NeoR/KanR	aminoglycoside phosphotransferase fromTn5, confers resistance to neomycin, kanamycin, andG418
10	BBa K2543009	SV40 poly(A) signal	SV40 polyadenylation signal
11	BBa_J64997	T7 consensus -10 and rest	This part has been characterized for GFP expression.
12	BBa K5487110	M13 rev	common sequencing primer, one of multiple similarvariants
13	BBa K4673024	lac operator	The lac repressor binds to the lac operator to inhibittranscription in E, coli. This inhibition can berelieved by adding lactose orisopropyl-β-D-thiogalactopyranoside(IPTG).
14	BBa K1804001	lac promoter	promoter for the E. coli lac operon
15	BBa M50499	CAP binding site	CAP binding activates transcription in the presenceof cAMP.
16	BBa I50022	ori	high-copy-number colE1/pMB1/pBR322/pUc originof replication
17	BBa K1927002	AmpR	confers resistance to ampicillin, carbenicillin, andrelated antibiotics
18	BBa K3766184	AmpR promoter	promoter for the AmpR
19	BBa K3830009	f1 ori	f1 bacteriophage origin of replication; arrowindicates direction of(+)strand synthesis

	Part number	Name	Brief introductions
1	BBa_K5487114	M13 fwd	common sequencing primer, one of multiple similar variants.
2	BBa_K2217024	CMV Enhancer + CMV Promoter	It is composed of CMV Enhancer as well as CMV Promoter (BBa_K2217000 and BBa_K2217006 respectively) as constitutive promoters to enhance the transcription process.
3	BBa_K4344028	Kozak sequence	The Kozak-sequence is the conserved consensus sequence next to the promoter, surrounding the AUG for the start of transcription in eukaryotes.
4	BBa_K5142041	FOLR2	the coding sequence (cds) of human folate receptor protein FOLR2
5	BBa_K1993019	T2A	T2A is a self-cleaving 2A peptide (The average length is 18–22 amino acids) that could be applied in expression of more than one gene in cells.
6	BBa_J06504	mCherry	the coding sequence (cds) of mCherry
7	BBa_K4818027	3xFLAG	The term "3xFLAG" refers to a protein tag used in molecular biology and biochemistry to facilitate the detection, purification, and characterization of proteins.
8	BBa K1722006	SV40 promoter	SV40 enhancer and early promoter
9	BBa K45740	NeoR/KanR	aminoglycoside phosphotransferase fromTn5, confers resistance to neomycin, kanamycin, andG418
10	BBa K2543009	SV40 poly(A) signal	SV40 polyadenylation signal
11	BBa_J64997	T7 consensus -10 and rest	This part has been characterized for GFP expression.
12	BBa K5487110	M13 rev	common sequencing primer, one of multiple similarvariants
13	BBa K4673024	lac operator	The lac repressor binds to the lac operator to inhibittranscription in E, coli. This inhibition can berelieved by adding lactose orisopropyl-β-D-thiogalactopyranoside(IPTG).
14	BBa K1804001	lac promoter	promoter for the E. coli lac operon
15	BBa M50499	CAP binding site	CAP binding activates transcription in the presenceof cAMP.
16	BBa I50022	ori	high-copy-number colE1/pMB1/pBR322/pUc originof replication
17	BBa K1927002	AmpR	confers resistance to ampicillin, carbenicillin, andrelated antibiotics
18	BBa K3766184	AmpR promoter	promoter for the AmpR
19	BBa K3830009	f1 ori	f1 bacteriophage origin of replication; arrowindicates direction of(+)strand synthesis