LOCUS Exported 5441 bp ds-DNA circular SYN 26-DEC-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..5441
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(4..819)
/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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin 941..1529
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 1715..1857
/label=bom
/note="basis of mobility region from pBR322"
CDS complement(1959..2150)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
promoter 3711..3729
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
RBS 3761..3783
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 3796..3813
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 3817..4917
/codon_start=1
/gene="malE (mutated)"
/product="maltose binding protein from E. coli"
/label=MBP
/note="This version of the gene does not encode a signal
sequence, so MBP will remain in the cytosol."
/translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
WYAVRTAVINAASGRQTVDEALKDAQT"
CDS 4969..4989
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
misc_feature 4991..5049
/label=MCS
CDS 5057..5074
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
terminator 5141..5188
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
ORIGIN
1 ttcttagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
61 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
121 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
181 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
241 gactgaatcc ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg
301 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
361 ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
421 cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg
481 atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc
541 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
601 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
661 aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc
721 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
781 cggcctcgag caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt
841 tatgtaagca gacagtttta ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc
901 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
961 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
1021 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
1081 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
1141 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
1201 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
1261 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
1321 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
1381 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
1441 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
1501 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
1561 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
1621 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
1681 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
1741 atctgtgcgg tatttcacac cgcatatatg gtgcactctc agtacaatct gctctgatgc
1801 cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc
1861 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
1921 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
1981 ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag
2041 atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg
2101 cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc
2161 gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc
2221 acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa
2281 ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc
2341 gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg
2401 aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg
2461 aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt
2521 cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg
2581 gtcctcaacg acaggagcac gatcatgcgc acccgtggcc aggacccaac gctgcccgag
2641 atgcgccgcg tgcggctgct ggagatggcg gacgcgatgg atatgttctg ccaagggttg
2701 gtttgcgcat tcacagttct ccgcaagaat tgattggctc caattcttgg agtggtgaat
2761 ccgttagcga ggtgccgccg gcttccattc aggtcgaggt ggcccggctc catgcaccgc
2821 gacgcaacgc ggggaggcag acaaggtata gggcggcgcc tacaatccat gccaacccgt
2881 tccatgtgct cgccgaggcg gcataaatcg ccgtgacgat cagcggtcca gtgatcgaag
2941 ttaggctggt aagagccgcg agcgatcctt gaagctgtcc ctgatggtcg tcatctacct
3001 gcctggacag catggcctgc aacgcgggca tcccgatgcc gccggaagcg agaagaatca
3061 taatggggaa ggccatccag cctcgcgtcg cgaacgccag caagacgtag cccagcgcgt
3121 cggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag
3181 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca
3241 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct
3301 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc
3361 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgacgctctc
3421 ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg ttgagcaccg
3481 ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc
3541 ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt
3601 ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc
3661 cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat taatacgact
3721 cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac tttaagaagg
3781 agatatacca tgaaacatca ccatcaccat caccccatga aaatcgaaga aggtaaactg
3841 gtaatctgga ttaacggcga taaaggctat aacggtctcg ctgaagtcgg taagaaattc
3901 gagaaagata ccggaattaa agtcaccgtt gagcatccgg ataaactgga agagaaattc
3961 ccacaggttg cggcaactgg cgatggccct gacattatct tctgggcaca cgaccgcttt
4021 ggtggctacg ctcaatctgg cctgttggct gaaatcaccc cggacaaagc gttccaggac
4081 aagctgtatc cgtttacctg ggatgccgta cgttacaacg gcaagctgat tgcttacccg
4141 atcgctgttg aagcgttatc gctgatttat aacaaagatc tgctgccgaa cccgccaaaa
4201 acctgggaag agatcccggc gctggataaa gaactgaaag cgaaaggtaa gagcgcgctg
4261 atgttcaacc tgcaagaacc gtacttcacc tggccgctga ttgctgctga cgggggttat
4321 gcgttcaagt atgaaaacgg caagtacgac attaaagacg tgggcgtgga taacgctggc
4381 gcgaaagcgg gtctgacctt cctggttgac ctgattaaaa acaaacacat gaatgcagac
4441 accgattact ccatcgcaga agctgccttt aataaaggcg aaacagcgat gaccatcaac
4501 ggcccgtggg catggtccaa catcgacacc agcaaagtga attatggtgt aacggtactg
4561 ccgaccttca agggtcaacc atccaaaccg ttcgttggcg tgctgagcgc aggtattaac
4621 gccgccagtc cgaacaaaga gctggcaaaa gagttcctcg aaaactatct gctgactgat
4681 gaaggtctgg aagcggttaa taaagacaaa ccgctgggtg ccgtagcgct gaagtcttac
4741 gaggaagagt tggcgaaaga tccacgtatt gccgccacta tggaaaacgc ccagaaaggt
4801 gaaatcatgc cgaacatccc gcagatgtcc gctttctggt atgccgtgcg tactgcggtg
4861 atcaacgccg ccagcggtcg tcagactgtc gatgaagccc tgaaagacgc gcagactaat
4921 tcgagctcga acaacaacaa caataacaat aacaacaacc ccatgagcga gaatctttat
4981 tttcagggcg ccatggacta atgaggtacc ggatccgaat tcgagctccg tcgacaagct
5041 tgcggccgca ctcgagcacc accaccacca ccactgagat ccggctgcta acaaagcccg
5101 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
5161 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gatatccaca
5221 ggacgggtgt ggtcgccatg atcgcgtagt cgatagtggc tccaagtagc gaagcgagca
5281 ggactgggcg gcggccaaag cggtcggaca gtgctccgag aacgggtgcg catagaaatt
5341 gcatcaacgc atatagcgct agcagcacgc catagtgact ggcgatgctg tcggaatgga
5401 cgatatcccg caagaggccc ggcagtaccg gcataaccaa g
//