LOCUS       Exported               10475 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  MoClo level 2 destination vector pL2-1, complete sequence.
ACCESSION   HQ728317
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10475)
  AUTHORS   Weber E, Engler C, Gruetzner R, Werner S, Marillonnet S.
  TITLE     A modular cloning system for standardized assembly of multigene 
            constructs
  JOURNAL   PLoS ONE 6 (2), E16765 (2011)
  PUBMED    21364738
REFERENCE   2  (bases 1 to 10475)
  AUTHORS   Weber E, Engler C, Gruetzner R, Werner S, Marillonnet S.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-DEC-2010) Icon Genetics GmbH, Weinbergweg 22, 
            Halle/Saale 06120, Germany
REFERENCE   3  (bases 1 to 10475)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10475
                     /organism="MoClo level 2 destination vector pL2-1"
                     /mol_type="other DNA"
                     /db_xref="taxon:993666"
     misc_feature    1..4
                     /note="fusion site; other site"
     misc_feature    23..47
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(170..758)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(929..1723)
                     /codon_start=1
                     /gene="aphA"
                     /product="aminoglycoside 3-phosphotransferase"
                     /function="kanamycin resistance"
                     /label=aphA
                     /protein_id="ADZ30886.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            complement(929..1723)
                     /gene="aphA"
                     /label=aphA
     promoter        complement(1724..1828)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1909..4693)
                     /direction=LEFT
                     /label=pVS1 ori
                     /note="pVS1 ori"
     rep_origin      2111..2305
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(2371..3438)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     CDS             complement(3872..4501)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    4765..4789
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    4862..4865
                     /note="fusion site; other site"
     CDS             5115..6023
                     /codon_start=1
                     /gene="crtE"
                     /product="geranylgeranyl pyrophosphate synthase"
                     /label=crtE
                     /note="GGPP"
                     /protein_id="ADZ30887.1"
                     /translation="MTVCAKKHVHLTRDAAEQLLADIDRRLDQLLPVEGERDVVGAAMR
                     EGALAPGKRIRPMLLLLTARDLGCAVSHDGLLDLACAVEMVHAASLILDDMPCMDDAKL
                     RRGRPTIHSHYGEHVAILAAVALLSKAFGVIADADGLTPLAKNRAVSELSNAIGMQGLV
                     QGQFKDLSEGDKPRSAEAILMTNHFKTSTLFCASMQMASIVANASSEARDCLHRFSLDL
                     GQAFQLLDDLTDGMTDTGKDSNQDAGKSTLVNLLGPRAVEERLRQHLQLASEHLSAACQ
                     HGHATQHFIQAWFDKKLAAVS"
     gene            5115..6023
                     /gene="crtE"
                     /label=crtE
     CDS             6033..6761
                     /codon_start=1
                     /gene="crtW"
                     /product="beta-carotene ketolase"
                     /label=crtW
                     /protein_id="ADZ30888.1"
                     /translation="MSAHALPKADLTATSLIVSGGIIAAWLALHVHALWFLDAAAHPIL
                     AIANFLGLTWLSVGLFIIAHDAMHGSVVPGRPRANAAMGQLVLWLYAGFSWRKMIVKHM
                     AHHRHAGTDDDPDFDHGGPVRWYARFIGTYFGWREGLLLPVIVTVYALILGDRWMYVVF
                     WPLPSILASIQLFVFGTWLPHRPGHDAFPDRHNARSSRISDPVSLLTCFHFGGYHHEHH
                     LHPTVPWWRLPSTRTKGDTA"
     gene            6033..6761
                     /gene="crtW"
                     /label=crtW
     regulatory      6389..6398
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             6783..7931
                     /codon_start=1
                     /gene="crtY"
                     /product="lycopene cyclase"
                     /label=crtY
                     /protein_id="ADZ30889.1"
                     /translation="MQPHYDLILVGAGLANGLIALRLQQQQPDMRILLIDAAPQAGGNH
                     TWSFHHDDLTESQHRWIAPLVVHHWPDYQVRFPTRRRKLNSGYFCITSQRFAEVLQRQF
                     GPHLWMDTAVAEVNAESVRLKKGQVIGARAVIDGRGYAANSALSVGFQAFIGQEWRLSH
                     PHGLSSPIIMDATVDQQNGYRFVYSLPLSPTRLLIEDTHYIDNATLDPECARQNICDYA
                     AQQGWQLQTLLREEQGALPITLSGNADAFWQQRPLACSGLRAGLFHPTTGYSLPLAVAV
                     ADRLSALDVFTSASIHHAITHFARERWQQQGFFRMLNRMLFLAGPADSRWRVMQRFYGL
                     PEDLIARFYAGKLTLTDRLRILSGKPPVPVLAALQAIMTTHR"
     gene            6783..7931
                     /gene="crtY"
                     /label=crtY
     CDS             7943..9421
                     /codon_start=1
                     /gene="crtI"
                     /product="phytoene dehydrogenase"
                     /label=crtI
                     /protein_id="ADZ30890.1"
                     /translation="MKPTTVIGAGFGGLALAIRLQAAGIPVLLLEQRDKPGGRAYVYED
                     QGFTFDAGPTVITDPSAIEELFALAGKQLKEYVELLPVTPFYRLCWESGKVFNYDNDQT
                     RLEAQIQQFNPRDVEGYRQFLDYSRAVFKEGYLKLGTVPFLSFRDMLRAAPQLAKLQAW
                     RSVYSKVASYIEDEHLRQAFSFHSLLVGGNPFATSSIYTLIHALEREWGVWFPRGGTGA
                     LVQGMIKLFQDLGGEVVLNARVSHMETTGNKIEAVHLEDGRRFLTQAVASNADVVHTYR
                     DLLSQHPAAVKQSNKLQTKRMSNSLFVLYFGLNHHHDQLAHHTVCFGPRYRELIDEIFN
                     HDGLAEDFSLYLHAPCVTDSSLAPEGCGSYYVLAPVPHLGTANLDWTVEGPKLRDRIFA
                     YLEQHYMPGLRSQLVTHRMFTPFDFRDQLNAYHGSAFSVEPVLTQSAWFRPHNRDKTIT
                     NLYLVGAGTHPGAGIPGVIGSAKATAGLMLEDLI"
     gene            7943..9421
                     /gene="crtI"
                     /label=crtI
     CDS             9457..10347
                     /codon_start=1
                     /gene="crtB"
                     /product="phytoene synthase"
                     /label=crtB
                     /protein_id="ADZ30891.1"
                     /translation="MAVGSKSFATASKLFDAKTRRSVLMLYAWCRHCDDVIDDQTLGFQ
                     ARQPALQTPEQRLMQLEMKTRQAYAGSQMHEPAFAAFQEVAMAHDIAPAYAFDHLEGFA
                     MDVREAQYSQLDDTLRYCYHVAGVVGLMMAQIMGVRDNATLDRACDLGLAFQLTNIARD
                     IVDDAHAGRCYLPASWLEHEGLNKENYAAPENRQALSRIARRLVQEAEPYYLSATAGLA
                     GLPLRSAWAIATAKQVYRKIGVKVEQAGQQAWDQRQSTTTPEKLTLLLAASGQALTSRM
                     RAHPPRPAHLWQRPL"
     gene            9457..10347
                     /gene="crtB"
                     /label=crtB