LOCUS       Exported                5522 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCR2.1-sacB, complete sequence.
ACCESSION   KC577242
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5522)
  AUTHORS   Rholl D, Bruckbauer S.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-FEB-2013) Microbiology Immunology and Pathology, 
            Colorado State University, IDRC at Foothills Campus, Campus Delivery
            0922, Fort Collins, CO 80523-0922, USA
REFERENCE   2  (bases 1 to 5522)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5522
                     /organism="Cloning vector pCR2.1-sacB"
                     /mol_type="other DNA"
                     /db_xref="taxon:1332674"
     promoter        complement(70..88)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(95..111)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      253..633
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             1001..1795
                     /codon_start=1
                     /gene="nptII"
                     /product="NptII"
                     /label=nptII
                     /note="kanamycin resistance protein"
                     /protein_id="AGL73303.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SRLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVIHGDACLPNIMVENGRFSGFNDCGRLGVADRYQDIA
                     LDTRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            1001..1795
                     /gene="nptII"
                     /label=nptII
     CDS             1813..2673
                     /codon_start=1
                     /gene="bla"
                     /product="Bla"
                     /label=bla
                     /note="ampicillin resistance protein"
                     /protein_id="AGL73304.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            1813..2673
                     /gene="bla"
                     /label=bla
     rep_origin      2841..3514
                     /label=pUC oriV
                     /note="pUC oriV"
     rep_origin      2844..3432
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    3720..3741
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3756..3786
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3794..3810
                     /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)."
     primer_bind     3818..3834
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3979..5439
                     /codon_start=1
                     /gene="sacB"
                     /product="SacB"
                     /label=sacB
                     /note="counter-selection marker on sucrose; orientation 
                     unknown"
                     /protein_id="AGL73302.1"
                     /translation="MTYSKQAVSRRHKRLALSAAALTAAACMSAHAQSDGATQAFAKET
                     NQKPYKETYGISHITRHDMLQIPEQQKNEKYQVSEFDSSTIKNISSAKGLDVWDSWPLQ
                     NADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFD
                     ANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNIN
                     GVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTE
                     DGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKV
                     MKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTG
                     PYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAP
                     SFLLNIKGKKTSVVKDSILEQGQLTVNK"
     gene            3979..5439
                     /gene="sacB"
                     /label=sacB
     terminator      5450..5495
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"