LOCUS       Exported               10639 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Chloroplast transformation vector pAXT, complete sequence.
ACCESSION   MF580999
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10639)
  AUTHORS   Lu Y, Stegemann S, Agrawal S, Karcher D, Ruf S, Bock R.
  TITLE     Horizontal Transfer of a Synthetic Metabolic Pathway between Plant 
            Species
  JOURNAL   Curr. Biol. (2017) In press
  PUBMED    28943084
REFERENCE   2  (bases 1 to 10639)
  AUTHORS   Karcher D, Bock R.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-AUG-2017) Organelle Biology, Biotechnology and 
            Molecular Ecophysiology, Max Planck Institute of Molecular Plant 
            Physiology, Am Muehlenberg 1, Potsdam, Brandenburg 14476, Germany
REFERENCE   3  (bases 1 to 10639)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10639
                     /organism="Chloroplast transformation vector pAXT"
                     /mol_type="other DNA"
                     /db_xref="taxon:2041179"
     promoter        36..54
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      73..201
                     /regulatory_class="promoter"
                     /note="Prrn"
     5UTR           202..243
     CDS             244..1758
                     /codon_start=1
                     /gene="Lyc"
                     /product="lycopene beta-cyclase"
                     /label=Lyc
                     /note="derived from daffodil"
                     /protein_id="ATG32096.1"
                     /translation="MDTLLRTHNRLELLYPLHELAKRHFLSPSPNPQNPNFKFFSRKPY
                     QKKCRNGYIGVSSNQLLDLVPEIKKEHLEFDLPLYDPSKALTLDLAVVGGGPAGLAVAQ
                     QVSEAGLSVVSIDPNPKLIWPNNYGVWVDEFEDMDLLDCLDATWSGAIVYVDDRSTKNL
                     SRPYARVNRKNLKSKMMKKCVSNGVRFHQATVVKAMHEEEKSYLICSDGVTIDARVVLD
                     ATGFSRCLVQYDKPYNPGYQVAYGILAEVEEHPFDVDKMVFMDWRDSHLNGKAELNERN
                     AKIPTFLYAMPFSSNRIFLEETSLVARPGLKMEDIQERMVARLNHLGIRIKSIEEDERC
                     VIPMGGPLPVIPQRVVGIGGTAGMVHPSTGYMVARTLAAAPIVANSIVQYLVSDSGLSG
                     NDLSADVWKDLWPIERRRQREFFCFGMDILLKLDLEGTRRFFDAFFDLEPRYWHGFLSS
                     RLFLPELVPFGLSLFSHASNTCKLEIMAKGTLPLVNMINNLVQDRD"
     gene            244..1758
                     /gene="Lyc"
                     /label=Lyc
                     /note="NpLyc"
     regulatory      1769..1918
                     /regulatory_class="terminator"
                     /note="derived from Nicotiana tabacum rps16"
     misc_feature    1931..1980
                     /note="IEE; intercistronic expression element"
     CDS             2005..2739
                     /codon_start=1
                     /gene="crtW"
                     /product="beta-carotene ketolase"
                     /label=crtW
                     /note="derived from marine bacterium Brevundimonas"
                     /protein_id="ATG32097.1"
                     /translation="MTAAVAEPRIVPRQTWIGLTLAGMIVAGWGSLHVYGVYFHRWGTS
                     SLVIVPAIVAVQTWLSVGLFIVAHDAMHGSLAPGRPRLNAAVGRLTLGLYAGFRFDRLK
                     TAHHAHHAAPGTADDPDFYAPAPRAFLPWFLNFFRTYFGWREMAVLTALVLIALFGLGA
                     RPANLLTFWAAPALLSALQLFTFGTWLPHRHTDQPFADAHHARSSGYGPVLSLLTCFHF
                     GRHHEHHLTPWRPWWRLWRGES"
     gene            2005..2739
                     /gene="crtW"
                     /label=crtW
                     /note="BsCrtW"
     regulatory      2746..2970
                     /regulatory_class="terminator"
                     /note="derived from Nicotiana tabacum rbcL"
     misc_feature    2976..3025
                     /note="IEE; intercistronic expression element"
     CDS             3045..3530
                     /codon_start=1
                     /gene="crtZ"
                     /product="beta-carotene hydroxylase"
                     /label=crtZ
                     /note="derived from marine bacterium Brevundimonas"
                     /protein_id="ATG32098.1"
                     /translation="MAWLTWIALFLTAFLGMEAFAWIMHRYVMHGFLWSWHRSHHEPHD
                     HPLEKNDLFAVVFAAPAIVMVAVGLHLWPWALPVGLGITAYGMVYFFFHDGLVHRRFPT
                     GFSGRSGFWTRRIQAHRLHHAVRTREGCVSFGFLWVRSARALKAELAQKRGSSSSGA"
     gene            3045..3530
                     /gene="crtZ"
                     /label=crtZ
                     /note="BsCrtZ"
     regulatory      3537..3758
                     /regulatory_class="terminator"
                     /note="derived from Chlamydomonas rbcL"
     misc_feature    complement(3792..3825)
                     /label=loxP
                     /note="loxP"
     protein_bind    3792..3825
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      complement(3826..4233)
                     /regulatory_class="terminator"
                     /note="derived from Nicotiana tabacum psbA"
     CDS             complement(4234..5025)
                     /codon_start=1
                     /gene="aadA"
                     /product="streptomycin/spectinomycin adenylyltransferase"
                     /label=aadA
                     /protein_id="ATG32095.1"
                     /translation="MAEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVLEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     gene            complement(4234..5025)
                     /gene="aadA"
                     /label=aadA
     regulatory      complement(5026..5200)
                     /regulatory_class="promoter"
                     /note="Prrn; derived from Nicotiana tabacum"
     misc_feature    complement(5201..5234)
                     /label=loxP
                     /note="loxP"
     protein_bind    5201..5234
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     tRNA            complement(5427..5497)
                     /gene="trnG"
                     /product="tRNA-Gly"
                     /label=trnG
     gene            complement(5427..5497)
                     /gene="trnG"
                     /label=trnG
     misc_feature    complement(5773..5903)
                     /label=similar to psbZ
                     /note="similar to psbZ"
     promoter        complement(5917..5935)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5956..5972)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5980..5996
                     /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)."
     promoter        complement(6004..6034)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6049..6070
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6358..6946)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7117..7977)
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase"
                     /label=ampR
                     /protein_id="ATG32100.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(7117..7977)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(7978..8082)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8108..8563)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     8705..8721
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8731..8749
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    8761..9961
                     /label=similar to psaB
                     /note="similar to psaB"
     CDS             10084..10386
                     /codon_start=1
                     /gene="rps14"
                     /product="ribosomal protein S14"
                     /label=rps14
                     /protein_id="ATG32099.1"
                     /translation="MARKSLIQREKKRQKLEQKYHSIRRSSKKEISKVPSLSDKWEIYG
                     KLQSPPRNSAPTRLHRRCFLTGRPRANYRDFGLSGHILREMVHACLLPGATRSSW"
     gene            10084..10386
                     /gene="rps14"
                     /label=rps14
     tRNA            10536..10609
                     /gene="trnfM"
                     /product="tRNA-fMet"
                     /label=trnfM
     gene            10536..10609
                     /gene="trnfM"
                     /label=trnfM