LOCUS       Exported                4342 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  BioBrick cloning vector pSB3T5-I52001, complete sequence.
ACCESSION   EU496104
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4342)
  AUTHORS   Shetty RP, Endy D, Knight TF Jr.
  TITLE     Engineering BioBrick vectors from BioBrick parts
  JOURNAL   J Biol Eng 2, 5 (2008)
  PUBMED    18410688
REFERENCE   2  (bases 1 to 4342)
  AUTHORS   Shetty RP, Endy D, Knight TF Jr.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-FEB-2008) Biological Engineering, MIT, 32 Vassar 
            Street Room 32-311, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 4342)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4342
                     /organism="BioBrick cloning vector pSB3T5-I52001"
                     /mol_type="other DNA"
                     /db_xref="taxon:508433"
     misc_feature    1..21
                     /label=BioBrick cloning site suffix (BBa_G00001)
                     /note="BioBrick cloning site suffix (BBa_G00001)"
     misc_feature    35..46
                     /note="translational stop sequence for BioBrick 
                     vector-insert insulation, has stop codons in all 6 reading 
                     frames (BBa_B0042)"
     regulatory      47..115
                     /regulatory_class="terminator"
                     /note="derived from tonB terminator (BBa_B0054); similar to
                     Escherichia coli strain MG1655 accession U00096"
     stem_loop       57..88
                     /note="derived from tonB terminator, predicted using mFold"
     terminator      57..88
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     primer_bind     complement(116..135)
                     /note="VR reverse sequencing primer annealing site 
                     (BBa_G00102)"
     regulatory      136..176
                     /regulatory_class="terminator"
                     /note="derived from ribosomal RNA rrnC operon, reverse 
                     orientation (BBa_B0062); similar to Escherichia coli strain
                     MG1655 accession U00096"
     stem_loop       complement(149..168)
                     /label=derived from ribosomal RNA rrnC operon
                     /note="derived from ribosomal RNA rrnC operon"
     rep_origin      184..1737
                     /note="p15A low to medium copy replication origin 
                     (BBa_I50032)"
     rep_origin      582..1406
                     /label=p15A replication origin
                     /note="p15A replication origin"
     rep_origin      599..1143
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     promoter        1759..1787
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             1835..3025
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline resistance protein"
                     /label=tet
                     /note="confers resistance to tetracycline"
                     /protein_id="ACB05500.1"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     gene            1835..3025
                     /gene="tet"
                     /label=tet
     regulatory      3036..3107
                     /regulatory_class="terminator"
                     /note="derived from hisI terminator (BBa_B0053); similar to
                     Escherichia coli strain MG1655 accession U00096"
     stem_loop       3044..3078
                     /label=derived from hisI terminator
                     /note="derived from hisI terminator"
     primer_bind     3108..3127
                     /note="VF2 forward sequencing primer annealing site 
                     (BBa_G00100)"
     regulatory      3128..3205
                     /regulatory_class="terminator"
                     /note="derived from bacteriophage T3 terminator, reverse 
                     orientation (BBa_B0055); similar to bacteriophage T3 gene 1
                     to gene 11 (accession number X17255)"
     terminator      3155..3184
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     stem_loop       complement(3163..3184)
     misc_feature    3206..3217
                     /note="translational stop sequence for BioBrick 
                     vector-insert insulation, has stop codons in all 6 reading 
                     frames (BBa_B0042)"
     misc_feature    3231..3252
                     /label=BioBrick cloning site prefix (BBa_G00000)
                     /note="BioBrick cloning site prefix (BBa_G00000)"
     CDS             3360..3665
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB"
                     /label=ccdB
                     /note="controlled cell death protein B; toxin protein that 
                     inhibits DNA gyrase which functions during DNA replication 
                     to catalyze negative supercoiling"
                     /protein_id="ACB05501.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            3360..3665
                     /gene="ccdB"
                     /label=ccdB
     misc_feature    3669..4342
                     /note="nonfunctional high copy replication origin derived 
                     from pUC19 accession M77789 (BBa_I50020)"
     gene            3695..4283
                     /gene="RNAII"
                     /label=RNAII
     rep_origin      3695..4283
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      3695..3700
                     /regulatory_class="minus_35_signal"
                     /gene="RNAII"
     regulatory      3716..3721
                     /regulatory_class="minus_10_signal"
                     /gene="RNAII"
     misc_RNA        3731..4283
                     /gene="RNAII"
                     /product="RNAII"
     gene            complement(3734..3877)
                     /gene="RNAI"
                     /label=RNAI
     misc_RNA        complement(3734..3841)
                     /gene="RNAI"
                     /product="RNAI"
     regulatory      complement(3850..3855)
                     /regulatory_class="minus_10_signal"
                     /gene="RNAI"
     regulatory      complement(3872..3877)
                     /regulatory_class="minus_35_signal"
                     /gene="RNAI"
     misc_difference 3929
                     /gene="RNAII"
                     /replace="g"
                     /note="G to A mutation to eliminate TspRI endonuclease 
                     site, renders BBa_I50020 nonfunctional as a replication 
                     origin"
     misc_difference 3933
                     /gene="RNAII"
                     /replace="g"
                     /note="G to A mutation to eliminate EcoP15I endonuclease 
                     site, renders BBa_I50020 nonfunctional as a replication 
                     origin"
     misc_difference 3942
                     /gene="RNAII"
                     /replace="g"
                     /note="G to A mutation to eliminate TspRI endonuclease 
                     site, renders BBa_I50020 nonfunctional as a replication 
                     origin"
     misc_difference 3971
                     /gene="RNAII"
                     /replace="c"
                     /note="C to T mutation to eliminate Nt.BstNBI endonuclease 
                     site, renders BBa_I50020 nonfunctional as a replication 
                     origin"
     misc_difference 4135
                     /gene="RNAII"
                     /replace="c"
                     /note="C to T mutation to eliminate BciVI endonuclease 
                     site, renders BBa_I50020 nonfunctional as a replication 
                     origin"