pECIA-2 vector (V007651)

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LOCUS       40924_16785        6007 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pECIA-2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6007)
  AUTHORS   Ozkan E, Carrillo RA, Eastman CL, Weiszmann R, Waghray D, Johnson 
            KG, Zinn K, Celniker SE, Garcia KC.
  TITLE     An Extracellular Interactome of Immunoglobulin and LRR Proteins 
            Reveals Receptor-Ligand Networks
  JOURNAL   Cell 154 (1), 228-239 (2013)
  PUBMED    23827685
REFERENCE   2  (bases 1 to 6007)
  AUTHORS   Ozkan E, Eastman CL, Garcia KC.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-JUL-2013) Molecular and Cellular Physiology, Stanford 
            University School of Medicine, 279 Campus Drive, Beckman Center, 
            Stanford, CA 94306, USA
REFERENCE   3  (bases 1 to 6007)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6007)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell"; 
            date: "2013"; volume: "154"; issue: "1"; pages: "228-239"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (24-JUL-2013) Molecular and Cellular Physiology, Stanford University
            School of Medicine, 279 Campus Drive, Beckman Center, Stanford, CA 
            94306, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6007
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        401..827
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     sig_peptide     845..898
                     /label=BiP signal sequence
                     /note="Drosophila BiP signal sequence"
     protein_bind    908..1032
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1057..1087
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1141..1797
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2142..2444
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2488..2612)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2638..2661
                     /codon_start=1
                     /label=HRV 3C site
                     /note="recognition and cleavage site for human rhinovirus
                     3C and PreScission proteases"
                     /translation="LEVLFQGP"
     misc_feature    2668..3348
                     /label=IgG1 heavy chain Fc fragment
                     /note="IgG1 heavy chain Fc fragment"
     CDS             3367..3408
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     CDS             3418..3435
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     polyA_signal    complement(3508..3642)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3786..3802)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3810..3826)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(3834..3864)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3879..3900)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4188..4776)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4950..5807)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5808..5912)
                     /label=AmpR promoter