LOCUS       Exported                9499 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  TREX vector pIC20H-RL, complete sequence.
ACCESSION   JX668229
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9499)
  AUTHORS   Loeschcke A, Markert A, Wilhelm S, Wirtz A, Rosenau F, Jaeger KE, 
            Drepper T.
  TITLE     TREX: A Universal Tool for the Transfer and Expression of 
            Biosynthetic Pathways in Bacteria
  JOURNAL   ACS Synth Biol 2 (1), 22-33 (2013)
  PUBMED    23656323
REFERENCE   2  (bases 1 to 9499)
  AUTHORS   Loeschcke A, Markert A, Drepper T, Jaeger K-E.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-SEP-2012) Institute of Molecular Enzyme Technology, 
            Heinrich-Heine-University, Stetternicher Forst, Juelich, NRW 52426, 
            Germany
REFERENCE   3  (bases 1 to 9499)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9499
                     /organism="TREX vector pIC20H-RL"
                     /mol_type="other DNA"
                     /db_xref="taxon:1256640"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        142..172
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(274..295)
                     /regulatory_class="promoter"
                     /note="PT7; T7 RNA polymerase-specific promoter"
     promoter        complement(274..292)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             326..1120
                     /codon_start=1
                     /gene="aphII"
                     /product="aminoglycoside 3-phosphotransferase"
                     /label=aphII
                     /note="kanamycin resistance"
                     /protein_id="AFW89950.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            326..1120
                     /gene="aphII"
                     /label=aphII
     misc_feature    1178..1196
                     /note="OE; outside end of transposon Tn5"
     oriT            1247..2438
                     /label=origin of conjuational transfer
                     /note="origin of conjuational transfer"
     gene            2537..3803
                     /gene="tetR"
                     /label=tetR
     promoter        2537..2565
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             2613..3803
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline export protein"
                     /label=tetR
                     /protein_id="AFW89947.1"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     misc_feature    complement(3861..3879)
                     /note="OE; outside end of transposon Tn5"
     gene            3890..5383
                     /gene="tnp"
                     /label=tnp
     CDS             3953..5383
                     /codon_start=1
                     /gene="tnp"
                     /product="transposase"
                     /label=tnp
                     /protein_id="AFW89948.1"
                     /translation="MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKS
                     ITISSEGSEAMQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYR
                     HQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESG
                     KWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGL
                     YLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNA
                     VLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAER
                     QRMEEPDNLERMVSILSFVAVRLLQLRESFTLPQALRAQGLLKEAEHVESQSAETVLTP
                     DECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQS
                     KLDGFLAAKDLMAQGIKI"
     gene            complement(5483..6233)
                     /gene="aacC1"
                     /label=aacC1
     CDS             complement(5483..6016)
                     /codon_start=1
                     /gene="aacC1"
                     /product="aminoglycoside acetyltransferase"
                     /label=aacC1
                     /note="getamicin resistance"
                     /protein_id="AFW89949.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(6205..6233)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             complement(6296..7015)
                     /codon_start=1
                     /gene="yfp"
                     /product="yellow fluorescent protein"
                     /label=yfp
                     /note="fluorescence reporter"
                     /protein_id="AFW89951.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     gene            complement(6296..7015)
                     /gene="yfp"
                     /label=yfp
     regulatory      7024..7045
                     /regulatory_class="promoter"
                     /note="PT7; T7 RNA polymerase-specific promoter"
     promoter        7027..7045
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    7046..7070
                     /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)."
     misc_feature    7050..7069
                     /label=lacO, lac operator
                     /note="lacO, lac operator"
     primer_bind     complement(7103..7119)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7593..7697
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7698..8558
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="ampicillin resistance"
                     /protein_id="AFW89952.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            7698..8558
                     /gene="bla"
                     /label=bla
     rep_origin      8729..9317
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"