LOCUS       Exported                8959 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pKRCPN1, complete sequence.
ACCESSION   KT991388
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8959)
  AUTHORS   Monson RE, Drew AM, Smith D, Matilla M, Roberts K, Richardson E, 
            Williamson N, Ramsay J, Welch M, Salmond GPC.
  TITLE     A plasmid-transposon hybrid mutagenesis system effective in a broad 
            range of Enterobacteria
  JOURNAL   Front Microbiol (2016) In press
REFERENCE   2  (bases 1 to 8959)
  AUTHORS   Monson RE, Drew AM, Smith D, Matilla M, Roberts K, Richardson E, 
            Williamson N, Ramsay J, Welch M, Salmond GPC.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-NOV-2015) Biochemistry, University of Cambridge, 
            Tennis Court Road, Cambridge CB2 1QW, United Kingdom
REFERENCE   3  (bases 1 to 8959)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8959
                     /organism="Cloning vector pKRCPN1"
                     /mol_type="other DNA"
                     /db_xref="taxon:1767046"
     CDS             complement(103..1293)
                     /codon_start=1
                     /gene="tetA"
                     /product="TetA"
                     /label=tetA
                     /note="tetracycline resistance"
                     /protein_id="ALS31074.1"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     gene            complement(103..1293)
                     /gene="tetA"
                     /label=tetA
     promoter        complement(1341..1369)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             1672..3177
                     /codon_start=1
                     /gene="tnp"
                     /product="transposase"
                     /label=tnp
                     /protein_id="ALS31075.1"
                     /translation="MSTNLSVIKNPRVQSDQRRLVRRPHMITSALHRAADWAKSVFSSA
                     ALGDPRRTARLVNVAAQLAKYSGKSITISSEGSKAAQEGAYRFIRNPNVSAEAIRKAGA
                     MQTVKLAQEFPELLAIEDTTSLSYRHQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFR
                     TVGLLHQEWWMRPDDPADADEKESGKWLAAAATSRLRMGSMMSNVIAVCDREADIHAYL
                     QDKLAHNERFVVRSKHPRKDVESGLYLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNR
                     PARKASLSLRSGRITLKQGNITLNAVLAEEINPPKGETPLKWLLLTSEPVESLAQALRV
                     IDIYTHRWRIEEFHKAWKTGAGAERQRMEEPDNLERMVSILSFVAVRLLQLRESFTPPQ
                     ALRAQGLLKEAEHVESQSAETVLTPDECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLG
                     GFMDSKRTGIASWGALWEGWEALQSKLDGFLAAKDLMAQGIKI"
     gene            1672..3177
                     /gene="tnp"
                     /label=tnp
     misc_feature    3186..3204
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     primer_bind     3431..3447
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3443..6517
                     /codon_start=1
                     /gene="lacZ"
                     /product="LacZ"
                     /label=lacZ
                     /note="beta-galactosidase"
                     /protein_id="ALS31076.1"
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYG
                     QDSRLPSEFDLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQ
                     ISDFHVATRFNDDFSRAVLEAEVQMCGKLRDYLRVTVSLWQGETQVASGTAPFGGEIID
                     ERGGYADRVTLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIEN
                     GLLLLNGKPLLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLW
                     YTLCDRYGLYVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWS
                     LGNESGHGANHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVP
                     KWSIKKWLSLPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSL
                     IKYDENGNPWSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQ
                     TIEVTSEYLFRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQ
                     LWLTVRVVQPNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELG
                     NKRWQFNRQSGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWK
                     AAGHYQAEAALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDV
                     EVASDTPHPARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPY
                     VFPSENGLRCGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDG
                     FHMGIGGDDSWSPSVSAELQLSAGRYHYQLVWCQK"
     gene            3443..6517
                     /gene="lacZ"
                     /label=lacZ
     primer_bind     complement(3463..3479)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6593..6981
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             7213..8028
                     /codon_start=1
                     /gene="aph"
                     /product="Aph"
                     /label=aph
                     /note="aminoglycoside 3-phosphotransferase"
                     /protein_id="ALS31077.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            7213..8028
                     /gene="aph"
                     /label=aph
     misc_feature    8390..8408
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     oriT            8647..8756
                     /note="incP origin of transfer"