LOCUS       Exported                5221 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBCSMH031, complete sequence.
ACCESSION   KC292052
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5221)
  AUTHORS   Henriques MX, Catalao MJ, Figueiredo J, Gomes JP, Filipe SR.
  TITLE     Construction of Improved Tools for Protein Localization Studies in 
            Streptococcus pneumoniae
  JOURNAL   PLoS ONE 8 (1), E55049 (2013)
  PUBMED    23349996
REFERENCE   2  (bases 1 to 5221)
  AUTHORS   Henriques MX, Catalao MJ, Filipe SR.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-DEC-2012) Bacterial Cell Surfaces and Pathogenesis, 
            Instituto de Tecnologia Quimica e Biologica - Universidade Nova de 
            Lisboa, Av. da Republica (EAN), Oeiras 2780-157, Portugal
REFERENCE   3  (bases 1 to 5221)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5221
                     /organism="Cloning vector pBCSMH031"
                     /mol_type="other DNA"
                     /label=derived from Streptococcus pneumoniae strain R36A
                     /note="derived from Streptococcus pneumoniae strain R36A"
                     /db_xref="taxon:1286973"
     regulatory      91..119
                     /regulatory_class="promoter"
                     /note="sigA-like"
     RBS             125..133
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3 end of the 16S rRNA (Chen et al., 1994)"
     regulatory      127..132
                     /regulatory_class="ribosome_binding_site"
     CDS             142..1020
                     /codon_start=1
                     /product="iCFP"
                     /label=iCFP
                     /note="linkers at N- and C-terminal ends"
                     /protein_id="AGD93427.1"
                     /translation="MPTLEIAQKKLELGTSSAGSAAGSGTSGPAGMVSKGEELFTGVVP
                     ILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTWGVQCFS
                     RYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFK
                     EDGNILGHKLEYNYISHNVYITADKQKNGIKANFKIRHNIEDGSVQLADHYQQNTPIGD
                     GPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYKAAAHMPWIRSQ
                     ELSDRPNDWLL"
     misc_feature    142..171
                     /label=i-Tag
                     /note="i-Tag"
     CDS             235..951
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             2700..2837
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A"
                     /label=repA
                     /protein_id="AGD93428.1"
                     /translation="MKKRLTITLSESVLENLEKMAREMGLSKSAMISVALENYKKGQEK
                     "
     gene            2700..2837
                     /gene="repA"
                     /label=repA
     CDS             2898..3530
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B"
                     /label=repB
                     /protein_id="AGD93429.1"
                     /translation="MAKEKARYFTFLLYPESIPSDWELKLETLGVPMAISPLHDKDKSS
                     IKGQKYKKAHYHVLYIAKNPVTADSVRKKIKLLLGEKSLAMVQVVLNVENMYLYLTHES
                     KDAIAKKKHVYDKADIKLINNFDIDRYVTLDVEEKTELFNVVVSLIRAYTLQNIFDLYD
                     FIDENGETYGLTINLVNEVIAGKTGFMKLLFDGAYQRSKRGTKNEER"
     gene            2898..3530
                     /gene="repB"
                     /label=repB
     CDS             3781..5157
                     /codon_start=1
                     /gene="Bacillus subtilis tetL"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MNTSYSQSNLRHNQILIWLCILSFFSVLNEMVLNVSLPDIANDFN
                     KPPASTNWVNTAFMLTFSIGTAVYGKLSDQLGIKRLLLFGIIINCFGSVIGFVGHSFFS
                     LLIMARFIQGAGAAAFPALVMVVVARYIPKENRGKAFGLIGSIVAMGEGVGPAIGGMIA
                     HYIHWSYLLLIPMITIITVPFLMKLLKKEVRIKGHFDIKGIILMSVGIVFFMLFTTSYS
                     ISFLIVSVLSFLIFVKHIRKVTDPFVDPGLGKNIPFMIGVLCGGIIFGTVAGFVSMVPY
                     MMKDVHQLSTAEIGSVIIFPGTMSVIIFGYIGGILVDRRGPLYVLNIGVTFLSVSFLTA
                     SFLLETTSWFMTIIIVFVLGGLSFTKTVISTIVSSSLKQQEAGAGMSLLNFTSFLSEGT
                     GIAIVGGLLSIPLLDQRLLPMEVDQSTYLYSNLLLLFSGIIVISWLVTLNVYKHSQRDF
                     "
     CDS             3871..5157
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline resistance protein"
                     /label=tet
                     /protein_id="AGD93426.1"
                     /translation="MVLNVSLPDIANDFNKPPASTNWVNTAFMLTFSIGTAVYGKLSDQ
                     LGIKRLLLFGIIINCFGSVIGFVGHSFFSLLIMARFIQGAGAAAFPALVMVVVARYIPK
                     ENRGKAFGLIGSIVAMGEGVGPAIGGMIAHYIHWSYLLLIPMITIITVPFLMKLLKKEV
                     RIKGHFDIKGIILMSVGIVFFMLFTTSYSISFLIVSVLSFLIFVKHIRKVTDPFVDPGL
                     GKNIPFMIGVLCGGIIFGTVAGFVSMVPYMMKDVHQLSTAEIGSVIIFPGTMSVIIFGY
                     IGGILVDRRGPLYVLNIGVTFLSVSFLTASFLLETTSWFMTIIIVFVLGGLSFTKTVIS
                     TIVSSSLKQQEAGAGMSLLNFTSFLSEGTGIAIVGGLLSIPLLDQRLLPMEVDQSTYLY
                     SNLLLLFSGIIVISWLVTLNVYKHSQRDF"
     gene            3871..5157
                     /gene="tet"
                     /label=tet