LOCUS       Exported                6885 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway vector p2RLGW7, complete sequence.
ACCESSION   KY434119
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6885)
  AUTHORS   Shen J, Liu J, Xie K, Xing F, Xiong F, Xiao J, Li X, Xiong L.
  TITLE     Translational repression by a miniature inverted-repeat transposable
            element in the 3 untranslated region
  JOURNAL   Nat Commun 8, 14651 (2017)
  PUBMED    28256530
REFERENCE   2  (bases 1 to 6885)
  AUTHORS   Shen J, Liu J, Xie K, Xing F, Xiong F, Xiao J, Li X, Xiong L.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-JAN-2017) Huazhong Agricultural University, National 
            Key Laboratory of Crop Genetic Improvement, Shizishan Street No. 1, 
            Wuhan, Hubei 430070, China
REFERENCE   3  (bases 1 to 6885)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6885
                     /organism="Gateway vector p2RLGW7"
                     /mol_type="other DNA"
                     /db_xref="taxon:1965291"
     promoter        complement(25..43)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(50..66)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      207..662
     promoter        740..844
                     /gene="bla"
                     /label=AmpR promoter
     CDS             845..1705
                     /codon_start=1
                     /product="beta lactamase"
                     /label=beta lactamase
                     /protein_id="AQX34622.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1876..2464
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2752..2773
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2788..2818
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2826..2842
                     /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     2850..2866
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2884..2902
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     regulatory      2950..3977
                     /regulatory_class="promoter"
                     /note="35S promoter"
     promoter        3562..3906
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             3999..4934
                     /codon_start=1
                     /product="renilla luciferase"
                     /label=renilla luciferase
                     /protein_id="AQX34623.1"
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     misc_feature    4941..5065
                     /note="recombination site attR1; other site"
     protein_bind    4941..5065
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        5090..5120
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             5174..5833
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=chloramphenicol acetyltransferase
                     /protein_id="AQX34624.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             6175..6480
                     /codon_start=1
                     /product="ccdB"
                     /label=ccdB
                     /protein_id="AQX34625.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     misc_feature    6521..6645
                     /note="recombination site attR2; other site"
     protein_bind    complement(6521..6645)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      6655..6880
                     /regulatory_class="terminator"
                     /note="35S terminator"