LOCUS       Exported               12139 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCPC52, complete sequence.
ACCESSION   MG874803
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12139)
  AUTHORS   Chang P, Wang W, Chen J.
  TITLE     Fast and convenient doxycycline-induced marker recycling in Candida 
            albicans via Cre-loxP system
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 12139)
  AUTHORS   Chang P, Wang W, Chen J.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-JAN-2018) Research Center of Bioenergy & 
            Bioremediation, Southwest University, 2 Tiansheng Road, Chongqiong, 
            Chongqing 400075, China
REFERENCE   3  (bases 1 to 12139)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12139
                     /organism="Cloning vector pCPC52"
                     /mol_type="other DNA"
                     /db_xref="taxon:2136745"
     CDS             262..1056
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=aminoglycoside phosphotransferase from Tn5
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (geneticin); NeoR/KanR"
                     /protein_id="AVR52682.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     regulatory      1257..1266
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1263..1637
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     rep_origin      1775..2363
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2651..2672
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2687..2717
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2725..2741
                     /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     2749..2765
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2783..2801
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     misc_feature    2881..4839
                     /label=5 region of CdARG4 gene
                     /note="5 region of CdARG4 gene"
     regulatory      4855..5589
                     /regulatory_class="promoter"
                     /note="ADH1 promoter from Candida albicans SC5314"
     CDS             5590..6573
                     /codon_start=1
                     /product="rTetR/GAL4AD fusion protein"
                     /label=rTetR/GAL4AD fusion protein
                     /note="Candida albicans codon-optimized"
                     /protein_id="AVR52683.1"
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGSTLFGGANFNQSGNI
                     ADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSNVHDNFMNNEITASKIDDGNNSKP
                     LSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDDEDTPPNPKKE"
     CDS             6229..6573
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     regulatory      6588..6894
                     /regulatory_class="terminator"
                     /note="ACT1 terminator from Candida albicans SC5314"
     CDS             7356..8252
                     /codon_start=1
                     /product="ATP phosphoribosyltransferase His1"
                     /label=ATP phosphoribosyltransferase His1
                     /note="HIS1 gene from Candida albicans SC5314; HIS1"
                     /protein_id="AVR52684.1"
                     /translation="MDLVNHLPDRLLFAVPKKGRLYEKCCNLLSGADIQFRRSNRLDIA
                     LSTNLPIALIFLPAADIPVFVGEGNCDLGITGLDQIKEAEQFDNIEDLLDLKFGSCKLQ
                     IQVPADGEYEKPEQLVGKKIVSSFTKLSTDYFKQLSDKPTNIRYVGGSVEASCALGVAD
                     AIVDLVESGETMKAAGLKAIETILETSAHLISSKKSKFPEMVNIIVQRLQGVLAAQEYV
                     LCNYNAPKSIQAKCLTITPGRRAATVSTLDKHSDDEEDWVAISSMVNRKEIGNVMDELK
                     KAGATDILVLEISNCRV"
     regulatory      8505..9296
                     /regulatory_class="promoter"
                     /note="tet promoter; TRE containing seven copies of the 
                     tetracycline operator tetO"
     protein_bind    8628..8898
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     CDS             9297..10328
                     /codon_start=1
                     /product="Cre recombinase"
                     /label=Cre recombinase
                     /note="Catalyzing of loxP sites recombination; 
                     codon-optimized for Candida albicans; Cre"
                     /protein_id="AVR52685.1"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     misc_feature    10535..11500
                     /label=3 region of ARG4 gene
                     /note="3 region of ARG4 gene"
     promoter        complement(11571..11589)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(11596..11612)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             11750..12052
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"