LOCUS       Exported                7277 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Plasmid vector pARAtrfA, complete sequence.
ACCESSION   JX524202
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7277)
  AUTHORS   Kvitko BH, McMillan IA, Schweizer HP.
  TITLE     An improved method for the oriT-directed cloning and 
            functionalization of large bacterial genomic regions
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7277)
  AUTHORS   Kvitko BH, McMillan IA, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-AUG-2012) Microbiology Immunology and Pathology, 
            Colorado State University, IDRC at Foothills Campus, Campus Delivery
            0922, Fort Collins, CO 80523-0922, USA
REFERENCE   3  (bases 1 to 7277)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7277
                     /organism="Plasmid vector pARAtrfA"
                     /mol_type="other DNA"
                     /label=RK2 replication helper vector
                     /note="RK2 replication helper vector"
                     /db_xref="taxon:1341207"
     CDS             complement(41..874)
                     /codon_start=1
                     /gene="rep-ts"
                     /product="temperature-sensitive Rep protein"
                     /label=rep-ts
                     /note="Rep-TS; pRO1600 Burkholderia variant"
                     /protein_id="AGN30565.1"
                     /translation="MASPPMVYKSNALVEAAYCLSVQEQRIVLASISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMGSTHAIRLYELLTQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKARKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     gene            complement(41..874)
                     /gene="rep-ts"
                     /label=rep-ts
     rep_origin      888..1239
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from 
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"
     promoter        1359..1406
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1425..1799
                     /codon_start=1
                     /gene="ble"
                     /product="zeocin resistance protein"
                     /label=ble
                     /note="Ble"
                     /protein_id="AGN30566.1"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     gene            1425..1799
                     /gene="ble"
                     /label=ble
     rep_origin      2077..2485
                     /label=M13 rep origin
                     /note="M13 rep origin"
     rep_origin      2080..2535
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      2451..3238
                     /label=pBR22 rep origin
                     /note="pBR22 rep origin"
     rep_origin      2646..3234
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3931..4860)
                     /codon_start=1
                     /gene="araC"
                     /product="AraC arabinose activator protein"
                     /label=araC
                     /note="pBAD promoter inducer"
                     /protein_id="AGN30567.1"
                     /translation="MQYGQLVSSLNGGSMKSMAEAQNDPLLPGYSFNAHLVAGLTPIEA
                     NGYLDFFIDRPLGMKGYILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPE
                     AREWYHQWVYFRPRAYWHEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGR
                     YSELLAINLLEQLLLRRMEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVC
                     LSPSRLSHLFRQQLGISVLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRV
                     FKKCTGASPSEFRAGCEEKVNDVAVKLS"
     gene            complement(3931..4860)
                     /gene="araC"
                     /label=araC
     CDS             complement(3931..4809)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4836..5120
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     protein_bind    5156..5180
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     RBS             5192..5212
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5222..6370
                     /codon_start=1
                     /gene="trfA1"
                     /product="replication initiation protein"
                     /label=trfA1
                     /note="TrfA1; acts on the RK2 (incP) plasmid origin"
                     /protein_id="AGN30568.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     gene            5222..6370
                     /gene="trfA1"
                     /label=trfA1
     protein_bind    complement(6387..6411)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      6655..6741
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6833..6860
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        6879..6970
                     /gene="bla"
                     /label=AmpR promoter