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"