LOCUS       Exported               10070 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pIREA, complete sequence.
ACCESSION   JX993919
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10070)
  AUTHORS   Wojcik EA, Brzostek A, Bacolla A, Mackiewicz P, Vasquez KM, 
            Korycka-Machala M, Jaworski A, Dziadek J.
  TITLE     Direct and inverted repeats elicit genetic instability by both 
            exploiting and eluding DNA double-strand break repair systems in 
            mycobacteria
  JOURNAL   PLoS ONE 7 (12), E51064 (2012)
  PUBMED    23251422
REFERENCE   2  (bases 1 to 10070)
  AUTHORS   Dziadek J, Wojcik EA.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-OCT-2012) Pharmacology and Toxicology, University of 
            Texas, 1400 Barbara Jordan Blvd., Austin, TX 78759, USA
REFERENCE   3  (bases 1 to 10070)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..10070
                     /organism="Cloning vector pIREA"
                     /mol_type="other DNA"
                     /label=pMV206-Km-Hyg-IR-GmA-lacZ LOCUS pMV206-Km-
                     /note="pMV206-Km-Hyg-IR-GmA-lacZ LOCUS pMV206-Km-"
                     /db_xref="taxon:1276270"
     misc_feature    6..1218
                     /label=aph1 region of Tn903
                     /note="aph1 region of Tn903"
     CDS             120..935
                     /codon_start=1
                     /gene="aphA1"
                     /product="aminoglycoside 3-phosphotransferase"
                     /EC_number="2.7.1.95"
                     /label=aphA1
                     /note="derived from Tn903 kanamycin resistance protein"
                     /protein_id="AGB57334.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPHAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHNLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            120..935
                     /gene="aphA1"
                     /label=aphA1
     rep_origin      complement(1223..2009)
                     /direction=LEFT
                     /label=Escherichia coli ColE1 (oriE) from pUC19
                     /note="Escherichia coli ColE1 (oriE) from pUC19"
     rep_origin      1267..1855
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(2028..3927)
                     /direction=LEFT
                     /label=mycobacterial OriM region from pAL5000
                     /note="mycobacterial OriM region from pAL5000"
     CDS             4192..7248
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /EC_number="3.2.1.23"
                     /label=lacZ
                     /note="hydrolysis of terminal non-reducing beta-D-galactose
                     residues in beta-D-galactosides"
                     /protein_id="AGB57332.1"
                     /translation="MDPVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQ
                     LRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITV
                     NPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLP
                     SEFDLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHV
                     ATRFNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYA
                     DRVTLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLN
                     GKPLLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDR
                     YGLYVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESG
                     HGANHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKK
                     WLSLPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDEN
                     GNPWSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTS
                     EYLFRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVR
                     VVQPNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQF
                     NRQSGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQ
                     AEAALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDT
                     PHPARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSEN
                     GLRCGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIG
                     GDDSWSPSVSAEFQLSAGRYHYQLVWCQK"
     gene            4192..7248
                     /gene="lacZ"
                     /label=lacZ
     CDS             4201..7248
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS
                     LNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNPP
                     FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF
                     DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR
                     FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV
                     TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP
                     LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL
                     YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA
                     NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS
                     LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP
                     WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL
                     FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ
                     PNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ
                     SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA
                     ALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP
                     ARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGLR
                     CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD
                     SWSPSVSAEFQLSAGRYHYQLVWCQK"
     CDS             complement(join(7510..8432,9363..9438))
                     /codon_start=1
                     /gene="hyg"
                     /product="hygromycin-B 7-O-kinase"
                     /EC_number="2.7.1.119"
                     /label=hyg
                     /note="hygromycin resistance"
                     /protein_id="AGB57333.1"
                     /translation="MTQESLLLLDRIDSDDSYASLRNDQEFWEPLARRALEELGLPVPP
                     VLRVPGESTNPVLVGEPDPVIKLFGEHWCGPESLASESEAYAVLADAPVPVPRLLGRGE
                     LRPGTGAWPWPYLVMSRMTGTTWRSAMDGTTDRNALLALARELGRVLGRLHRVPLTGNT
                     VLTPHSEVFPELLRERRAATVEDHRGWGYLSPRLLDRLEDWLPDVDTLLAGREPRFVHG
                     DLHGTNIFVDLAATEVTGIVDFTDVYAGDSRYSLVQLHLNAFRGDREILAALLDGAQWK
                     RTEDFARELLAFTFLHDFEVFEETPLDLSGFTDPEELAQFLWGPPDTAPGA"
     gene            complement(7510..9438)
                     /gene="hyg"
                     /label=hyg
                     /note="derived from Streptomyces hygroscopicus"
     repeat_region   8411..8450
                     /rpt_family="IR-1"
     CDS             complement(8568..9101)
                     /codon_start=1
                     /gene="aacC1"
                     /product="aminoglycoside N(3)-acetyltransferase I"
                     /EC_number="2.3.1.60"
                     /label=aacC1
                     /note="gentamicin resistance"
                     /protein_id="AGB57335.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            complement(8568..9101)
                     /gene="aacC1"
                     /label=aacC1
                     /note="gentamicin resistance"
     promoter        complement(9290..9318)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     repeat_region   9343..9378
                     /rpt_family="IR-2"
     terminator      9988..10034
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      complement(9991..10034)
                     /regulatory_class="terminator"
                     /note="rrnBT1"