LOCUS       Exported                5224 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBCSMH032, complete sequence.
ACCESSION   KC292053
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5224)
  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 5224)
  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 5224)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: Lasergene SeqMan v. 8.1.3; Lasergene
                                     SeqBuilder v. 8.1.3
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5224
                     /organism="Cloning vector pBCSMH032"
                     /mol_type="other DNA"
                     /label=derived from Streptococcus pneumoniae strain R36A
                     /note="derived from Streptococcus pneumoniae strain R36A"
                     /db_xref="taxon:1286974"
     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..1023
                     /codon_start=1
                     /product="iGFP"
                     /label=iGFP
                     /note="linkers at N- and C-terminal ends"
                     /protein_id="AGD93430.1"
                     /translation="MPTLEIAQKKLELGTSSAGSAAGSGTSGPAGMSKGEELFTGVVPI
                     LVELDGDVNGHRFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLAYGLQCFAR
                     YPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKE
                     DGNILGHKLEYNYNSHNVYIKADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDG
                     PVLLPDNHYLSTQSVLSKDPNEKRDHMILLEFVTAAGITHGMDELYKVDAAAHMPWIRS
                     QELSDRPNDWLL"
     misc_feature    142..172
                     /label=i-Tag
                     /note="i-Tag"
     CDS             235..948
                     /codon_start=1
                     /product="Aequoria victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIKADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMILLE
                     FVTAAGITHGMDELYK"
     CDS             2703..2840
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A"
                     /label=repA
                     /protein_id="AGD93431.1"
                     /translation="MKKRLTITLSESVLENLEKMAREMGLSKSAMISVALENYKKGQEK
                     "
     gene            2703..2840
                     /gene="repA"
                     /label=repA
     CDS             2901..3533
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B"
                     /label=repB
                     /protein_id="AGD93432.1"
                     /translation="MAKEKARYFTFLLYPESIPSDWELKLETLGVPMAISPLHDKDKSS
                     IKGQKYKKAHYHVLYIAKNPVTADSVRKKIKLLLGEKSLAMVQVVLNVENMYLYLTHES
                     KDAIAKKKHVYDKADIKLINNFDIDRYVTLDVEEKTELFNVVVSLIRAYTLQNIFDLYD
                     FIDENGETYGLTINLVNEVIAGKTGFMKLLFDGAYQRSKRGTKNEER"
     gene            2901..3533
                     /gene="repB"
                     /label=repB
     CDS             3784..5160
                     /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             3874..5160
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline resistance protein"
                     /label=tet
                     /protein_id="AGD93433.1"
                     /translation="MVLNVSLPDIANDFNKPPASTNWVNTAFMLTFSIGTAVYGKLSDQ
                     LGIKRLLLFGIIINCFGSVIGFVGHSFFSLLIMARFIQGAGAAAFPALVMVVVARYIPK
                     ENRGKAFGLIGSIVAMGEGVGPAIGGMIAHYIHWSYLLLIPMITIITVPFLMKLLKKEV
                     RIKGHFDIKGIILMSVGIVFFMLFTTSYSISFLIVSVLSFLIFVKHIRKVTDPFVDPGL
                     GKNIPFMIGVLCGGIIFGTVAGFVSMVPYMMKDVHQLSTAEIGSVIIFPGTMSVIIFGY
                     IGGILVDRRGPLYVLNIGVTFLSVSFLTASFLLETTSWFMTIIIVFVLGGLSFTKTVIS
                     TIVSSSLKQQEAGAGMSLLNFTSFLSEGTGIAIVGGLLSIPLLDQRLLPMEVDQSTYLY
                     SNLLLLFSGIIVISWLVTLNVYKHSQRDF"
     gene            3874..5160
                     /gene="tet"
                     /label=tet