LOCUS       Exported                5246 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pETL7, complete sequence.
ACCESSION   HQ007050
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5246)
  AUTHORS   Koksharov MI, Ugarova NN.
  TITLE     Thermostabilization of luciferase by in vivo directed evolution
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5246)
  AUTHORS   Koksharov MI, Ugarova NN.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-JUL-2010) Department of Chemistry, Lomonosov Moscow 
            State University, Leninskie Gory, Bld. 1/11, GSP-3, Moscow 119991, 
            Russia
REFERENCE   3  (bases 1 to 5246)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5246
                     /organism="Cloning vector pETL7"
                     /mol_type="other DNA"
                     /db_xref="taxon:910447"
     regulatory      21..39
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             71..93
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             101..1783
                     /codon_start=1
                     /product="N- and C-terminally tagged firefly luciferase"
                     /label=N- and C-terminally tagged firefly luciferase
                     /note="Luc; derived from Luciola mingrelica firefly 
                     luciferase in INSD accession S61961; contains additional 
                     MASK peptide at the N-terminus; native C-terminal peptide 
                     AKM is changed to SGPVEHHHHHH; contains 3 additional 
                     restriction sites (BstBI, SalI, XhoI) within the luciferase
                     gene"
                     /protein_id="ADP02956.1"
                     /translation="MASKMEMEKEENVVYGPLPFYPIEEGSAGIQLHKYMHQYAKLGAI
                     AFSNALTGVDISYQEYFDITCRLAEAMKNFGMKPEEHIALCSENCEEFFIPVLAGLYIG
                     VAVAPTNEIYTLRELNHSLGIAQPTIVFSSRKGLPKVLEVQKTVTCIKKIVILDSKVNF
                     GGHDCMETFIKKHVELGFQPSSFVPIDVKNRKQHVALLMNSSGSTGLPKGVRITHEGAV
                     TRFSHAKDPIYGNQVSPGTAILTVVPFHHGFGMFTTLGYFACGYRVVMLTKFDEELFLR
                     TLQDYKCTSVILVPTLFAILNKSELIDKFDLSNLTEIASGGAPLAKEVGEAVARRFNLP
                     GVRQGYGLTETTSAFIITPEGDDKPGASGKVVPLFKVKVIDLDTKKTLGVNRRGEICVK
                     GPSLMLGYSNNPEATRETIDEEGWLHTGDIGYYDEDEHFFIVDRLKSLIKYKGYQVPPA
                     ELESVLLQHPNIFDAGVAGVPDPDAGELPGAVVVMEKGKTMTEKEIVDYVNSQVVNHKR
                     LRGGVRFVDEVPKGLTGKIDAKVIREILKKPQSGPVEHHHHHH"
     CDS             1763..1780
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1847..1894
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      1848..1894
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     rep_origin      1931..2386
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      1948..2254
                     /label=f1 origin
                     /note="f1 origin"
     promoter        2413..2517
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      2448..2476
                     /regulatory_class="promoter"
                     /note="amp promoter"
     CDS             2518..3378
                     /codon_start=1
                     /product="Bla"
                     /label=Bla
                     /note="beta-lactamase; ampicillin resistance protein"
                     /protein_id="ADP02957.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3533..4152
                     /label=pBR322 origin
                     /note="pBR322 origin"
     rep_origin      3549..4137
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    4323..4465
                     /label=bom
                     /note="basis of mobility region from pBR322"
     misc_feature    complement(4567..4758)
                     /label=Region: ROP
                     /note="Region: ROP"
     CDS             complement(4567..4758)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"