LOCUS       Exported                6883 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTLR7-A-SEAP, complete sequence.
ACCESSION   KX035095
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6883)
  AUTHORS   Mejia JE, Azar P, Guery J-C.
  TITLE     Functional study of the rs179008 single-nucleotide polymorphism of 
            human TLR7
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6883)
  AUTHORS   Mejia JE, Azar P, Guery J-C.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-APR-2016) Centre de Physiopathologie de 
            Toulouse-Purpan, Inserm, CHU Purpan, BP 3028, Toulouse Cedex 3 
            31024, France
REFERENCE   3  (bases 1 to 6883)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6883
                     /organism="Cloning vector pTLR7-A-SEAP"
                     /mol_type="other DNA"
                     /db_xref="taxon:1932888"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1124..2302
                     /regulatory_class="promoter"
                     /note="EEF1A1; derived from human EF-1 alpha constitutive 
                     promoter and 5UTR"
     promoter        1124..2302
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1355..2293
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     5UTR           2308..2376
                     /label=derived from human TLR7
                     /note="derived from human TLR7"
     CDS             2377..3969
                     /codon_start=1
                     /product="TLR7/SEAP fusion protein"
                     /label=TLR7/SEAP fusion protein
                     /note="derived from secreted alkaline phosphatase and human
                     toll-like receptor 7 allele A rs179008"
                     /protein_id="APT68416.1"
                     /translation="MVFPMWTLKRQILILFNIILISKLLGARWFPVEEENPDFWNREAA
                     EALGAAKKLQPAQTAAKNLIIFLGDGMGVSTVTAARILKGQKKDKLGPEIPLAMDRFPY
                     VALSKTYNVDKHVPDSGATATAYLCGVKGNFQTIGLSAAARFNQCNTTRGNEVISVMNR
                     AKKAGKSVGVVTTTRVQHASPAGTYAHTVNRNWYSDADVPASARQEGCQDIATQLISNM
                     DIDVILGGGRKYMFRMGTPDPEYPDDYSQGGTRLDGKNLVQEWLAKRQGARYVWNRTEL
                     MQASLDPSVTHLMGLFEPGDMKYEIHRDSTLDPSLMEMTEAALRLLSRNPRGFFLFVEG
                     GRIDHGHHESRAYRALTETIMFDDAIERAGQLTSEEDTLSLVTADHSHVFSFGGYPLRG
                     SSIFGLAPGKARDRKAYTVLLYGNGPGYVLKDGARPDVTESESGSPEYRQQSAVPLDEE
                     THAGEDVAVFARGPQAHLVHGVQEQTFIAHVMAFAACLEPYTACDLAPPAGTTDAAHPG
                     YSRVGAAGRFEQT"
     regulatory      3961..4182
                     /regulatory_class="other"
                     /note="polyadenylation signal from SV40"
     polyA_signal    3970..4091
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    4239..688
                     /label=derived from pBC KS+
                     /note="derived from pBC KS+"
     promoter        complement(4256..4274)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4295..4311)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4319..4335
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(4343..4373)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4388..4409
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      4697..5285
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5674..6333)
                     /codon_start=1
                     /product="chloramphenicol acetyl transferase"
                     /label=chloramphenicol acetyl transferase
                     /note="confers resistance to chloramphenicol"
                     /protein_id="APT68415.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(6334..6436)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     promoter        complement(6756..6860)
                     /gene="bla"
                     /label=AmpR promoter