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miniTn4001-Puro-1 vector (V009844#)

Basic Vector Information

Basic Vector Information
Vector Name miniTn4001-Puro-1 Antibiotic Resistance Ampicillin
Length 5311 bp Type Cloning vector
Source Paetzold B, Carolis C, Ferrar T, Serrano L, Lluch-Senar M.

miniTn4001-Puro-1 vector Vector Map

SK primer T3 promoter M13 rev lac operator lac promoter CAP binding site ori beta lactamase AmpR promoter f1 ori M13 fwd T7 promoter transposase KS primer vertebrate consensus sequence for strong initiation of translation (Kozak, 1987) puromycin N-acetyltransferase

miniTn4001-Puro-1 vector Sequence

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LOCUS       Exported                5311 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector miniTn4001-Puro-1, complete sequence.
ACCESSION   KC816623
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5311)
  AUTHORS   Paetzold B, Carolis C, Ferrar T, Serrano L, Lluch-Senar M.
  TITLE     In situ overlap and sequence synthesis during DNA assembly
  JOURNAL   ACS Synth Biol 2 (12), 750-755 (2013)
  PUBMED    24161008
REFERENCE   2  (bases 1 to 5311)
  AUTHORS   Paetzold B.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-MAR-2013) EMBL-CRG Systems Biology Research Unit, 
            Centre for Genomic Regulation, Dr. Aiguader 88, Barcelona 08003, 
            Spain
REFERENCE   3  (bases 1 to 5311)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5311
                     /organism="Cloning vector miniTn4001-Puro-1"
                     /mol_type="other DNA"
                     /db_xref="taxon:1471065"
     primer_bind     complement(2..18)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     mobile_element  complement(27..52)
                     /mobile_element_type="insertion sequence:IS256"
     promoter        complement(74..92)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(113..129)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    137..153
                     /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(161..191)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    206..227
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(515..1103)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1274..2134)
                     /codon_start=1
                     /product="beta lactamase"
                     /EC_number="3.5.2.6"
                     /label=beta lactamase
                     /note="ampicillin resistance protein"
                     /protein_id="AHL28655.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2135..2239)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2265..2720)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     2862..2878
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2888..2906
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3008..4180
                     /codon_start=1
                     /product="transposase"
                     /label=transposase
                     /protein_id="AHL28656.1"
                     /translation="MTQVHFTLKSEEIQSIIEYSVKDDVSKNILTTVFNQLMENQRTEY
                     IQAKEYERTENRQSQRNGYYERSFTTRVGTLELKVPRTRDGHFSPTVFERYQRNEKALM
                     ASMLEMYVSGVSTRKVSKIVEELCGKSVSKSFVSSLTEQLEPMVNEWQNRLLSEKNYPY
                     LMTDVLYIKVREENRVLSKSCHIAIGITKDGDREIIGFMIQSGESEETWTTFFEYLKER
                     GLQGTELVISDAHKGLVSAIRKSFTNVSWQRCQVHFLRNIFTTIPKKNSKSFREAVKGI
                     FKFTDINLAREAKNRLIHDYIDQPKYSKACASLDDGFEDAFQYTVQGNSHNRLKSTNLI
                     ERLNQEVRRREKIIRIFPNQTSANRLIGAVLMDLHDEWIYSSRKYINFDK"
     mobile_element  4185..4210
                     /mobile_element_type="insertion sequence:IS256"
     primer_bind     4222..4238
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      4287..4296
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4627..5226
                     /codon_start=1
                     /product="puromycin N-acetyltransferase"
                     /EC_number="2.3.-.-"
                     /label=puromycin N-acetyltransferase
                     /protein_id="AHL28657.1"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"

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