LOCUS Exported 3754 bp ds-DNA circular SYN 14-DEC-2017
DEFINITION Gateway(R) Dual Selection Vector for creating an entry vector by
restriction cloning. Other reading frames are available with
pENTR(TM)2B and pENTR(TM)3C.
COMMENT To create a Kozak sequence, clone a blunt fragment starting with
ATGG (where ATG is the start codon) into the XmnI site.
FEATURES Location/Qualifiers
source 1..3754
/organism="synthetic DNA construct"
/lab_host="Escherichia coli"
/mol_type="other DNA"
terminator 103..189
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 281..308
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
protein_bind 358..457
/gene="mutant version of attL"
/label=attL1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
misc_feature 457..507
/label=MCS 1
/note="multiple cloning site, part 1"
RBS 461..466
/note="ribosome binding site"
promoter 524..554
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 608..1288
/codon_start=1
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
DPAY"
CDS 1608..1913
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
misc_feature 1948..1979
/label=MCS 2
/note="multiple cloning site, part 2"
protein_bind complement(1983..2082)
/gene="mutant version of attL"
/label=attL2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
CDS 2205..3014
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin 3104..3692
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
421 ttttttataa tgccaacttt gtacaaaaaa gcaggcttta aaggaaccaa ttcagtcgac
481 tggatccggt accgaattcg cggccgcatt aggcacccca ggctttacac tttatgcttc
541 cggctcgtat aatgtgtgga ttttgagtta ggatccgtcg agattttcag gagctaagga
601 agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg
661 taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca
721 gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc
781 ctttattcac attcttgccc gcctgatgaa tgctcatccg gaattccgta tggcaatgaa
841 agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca
901 aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca
961 catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt
1021 tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt
1081 aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac
1141 gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg
1201 cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcaggcggg
1261 gcgtaatcta gaggatccgg cttactaaaa gccagataac agtatgcgta tttgcgcgct
1321 gatttttgcg gtataagaat atatactgat atgtataccc gaagtatgtc aaaaagaggt
1381 atgctatgaa gcagcgtatt acagtgacag ttgacagcga cagctatcag ttgctcaagg
1441 catatatgat gtcaatatct ccggtctggt aagcacaacc atgcagaatg aagcccgtcg
1501 tctgcgtgcc gaacgctgga aagcggaaaa tcaggaaggg atggctgagg tcgcccggtt
1561 tattgaaatg aacggctctt ttgctgacga gaacaggggc tggtgaaatg cagtttaagg
1621 tttacaccta taaaagagag agccgttatc gtctgtttgt ggatgtacag agtgatatta
1681 ttgacacgcc cgggcgacgg atggtgatcc ccctggccag tgcacgtctg ctgtcagata
1741 aagtcccccg tgaactttac ccggtggtgc atatcgggga tgaaagctgg cgcatgatga
1801 ccaccgatat ggccagtgtg ccggtctccg ttatcgggga agaagtggct gatctcagcc
1861 accgcgaaaa tgacatcaaa aacgccatta acctgatgtt ctggggaata taaatgtcag
1921 gctcccttat acacagccag tctgcaggtc gacgaattcg cggccgcact cgagatatct
1981 agacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg
2041 caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag ctgcagctct
2101 ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa
2161 caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac
2221 gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
2281 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg
2341 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
2401 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
2461 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc
2521 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgtccc
2581 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
2641 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
2701 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat
2761 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
2821 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
2881 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
2941 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
3001 atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ttattcagat tgggccccgt
3061 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
3121 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
3181 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
3241 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
3301 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
3361 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
3421 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
3481 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
3541 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
3601 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
3661 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
3721 tcctggcctt ttgctggcct tttgctcaca tgtt
//