LOCUS Exported 10933 bp ds-DNA circular SYN 22-DEC-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..10933
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 36..1204
/label=nmt1 promoter
protein_bind 1238..1362
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 1387..1417
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 1471..2151
/codon_start=1
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
DPAY"
CDS 2471..2776
/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"
protein_bind complement(2817..2941)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
CDS 2953..2976
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 2983..3006
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 3007..3024
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
terminator 3069..3256
/gene="S. cerevisiae ADH1"
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
CDS complement(4920..5714)
/codon_start=1
/gene="Schizosaccharomyces pombe ura4"
/product="orotidine 5'-phosphate decarboxylase, required
for uracil biosynthesis"
/label=S. pombe ura4
/translation="MDARVFQSYSARAEGMKNPIAKELLALMEEKQSNLSVAVDLTKKS
EILELVDKIGPYVCVIKTHIDVVEDFDQDMVEKLVALGKKHRFLIFEDRKFADIGNTVK
LQYASGVYKIASWAHITNCHTVPGEGIIQGLKEVGLPLGRGLLLLAEMSSKGSLATGSY
TEKTLEWFEKHTDFCFGFIAGRRFPNLQSDYITMSPGIGLDVKGDGLGQQYRTPEEVIV
NCGSDIIIVGRGVYGAGRNPVVEAKRYREAGWKAYQQRLSQH"
rep_origin 6323..7106
/label=ars1
/note="Schizosaccharomyces pombe autonomously replicating
sequence ars1"
rep_origin 8058..8513
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 8795..8899
/gene="bla"
/label=AmpR promoter
CDS 8900..9760
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 9931..10519
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 aagcttgcat gcctgcaggt cgatcgactc tagaggatca gaaaattatc gccataaaag
61 acagaataag tcatcagcgg ttgtttcatt tcctatattt tttttttatt tttttatttt
121 ttaataaggg aaaatttaac gtctaaggat acagaagatt gttagcacat taaagtaata
181 aaggcttaag tagtaagtgc cttagcatgt tattgtattt caaaggacat aatctaaaat
241 aataacaata tcatttctca caagttattc aattttcttt tttttttcta ataatatcaa
301 gaatgtatta tttgtttgac ataagtcaac taatttattt aatatgctgg attaatcttg
361 cagacatgta aattaacaag ttttagtcaa ataacgttga agtttcaatg aactcaaata
421 atttctcttt ttttttatat aactctaatc tgatttatat tttccgcagg gatcaactga
481 agttatgaca tttggattgg atcacttata accttggtcg ccaaataata caaaaatcag
541 cgttataaaa caaagaaggt ttttgttaag aaattaatcc tctttcttga taagaaagtt
601 gaaccgaaat tgcagatact gatatatgaa aataataccc acaattttgg gaatagcgca
661 agcctcaatt taaacaatag gtgaggacac atgataatga cctcaatgat tgttagaaga
721 aaagagcctc attacaaaat cgaaaaatga atggttgggt acaagtttcc aaaacatggt
781 aaagtggact ttgcgtatga gacgtaaata gaaaaaaaca cttgttatat gttttctaga
841 attattgttg tctctttatg gttggatgat gcaaaatagt aatttcggtt agttgctgta
901 aaacaccacg agacaaatag atatggatat ttattaaatc aggaaaaacg taactctcgg
961 ctactggatg gttcagtcac ccaacgatta ctggggagag aaaacagggc aaaagcaaag
1021 cttaaaggaa tccgattgtc attcggcaat gtgcagcgaa actaaaaacc ggataatgga
1081 cctgttaatc gaaacattga agatatataa aggaagagga atcctggcat atcatcaatt
1141 gaataagttg aattaattat ttcaatctca ttctcacttt ctgacttata gtcgctttgt
1201 taaagctagc gtcgacaggc ctggatccgg atatcaaaca agtttgtaca aaaaagctga
1261 acgagaaacg taaaatgata taaatatcaa tatattaaat tagattttgc ataaaaaaca
1321 gactacataa tactgtaaaa cacaacatat ccagtcacta tggcggccgc attaggcacc
1381 ccaggcttta cactttatgc ttccggctcg tataatgtgt ggattttgag ttaggatccg
1441 tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc
1501 gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa
1561 tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa
1621 aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat
1681 ccggaattcc gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct
1741 tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac
1801 gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac
1861 ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg
1921 gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt
1981 ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag
2041 gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag
2101 tactgcgatg agtggcaggc ggggcgtaat ctagaggatc cggcttacta aaagccagat
2161 aacagtatgc gtatttgcgc gctgattttt gcggtataag aatatatact gatatgtata
2221 cccgaagtat gtcaaaaaga ggtatgctat gaagcagcgt attacagtga cagttgacag
2281 cgacagctat cagttgctca aggcatatat gatgtcaata tctccggtct ggtaagcaca
2341 accatgcaga atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa
2401 gggatggctg aggtcgcccg gtttattgaa atgaacggct cttttgctga cgagaacagg
2461 ggctggtgaa atgcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt
2521 tgtggatgta cagagtgata ttattgacac gcccgggcga cggatggtga tccccctggc
2581 cagtgcacgt ctgctgtcag ataaagtccc ccgtgaactt tacccggtgg tgcatatcgg
2641 ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggtct ccgttatcgg
2701 ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat
2761 gttctgggga atataaatgt caggctccct tatacacagc cagtctgcag gtcgaccata
2821 gtgactggat atgttgtgtt ttacagtatt atgtagtctg ttttttatgc aaaatctaat
2881 ttaatatatt gatatttata tcattttacg tttctcgttc agctttcttg tacaaagtgg
2941 ttcgatatca tggattataa ggatgatgac gataaaccca tggactacaa ggacgacgat
3001 gataaacatc accaccatca ccactgagat ctcgactcta gaggatccgg cgcgccactt
3061 ctaaataagc gaatttctta tgatttatga tttttattat taaataagtt ataaaaaaaa
3121 taagtgtata caaattttaa agtgactctt aggttttaaa acgaaaattc ttattcttga
3181 gtaactcttt cctgtaggtc aggttgcttt ctcaggtata gtatgaggtc gctcttattg
3241 accacacctc taccggcaga tccgctaggg ataacagggt aatatactag taagatgatt
3301 cttttatcgt cgataacgat gaggatgtag acgattatga tgagagtgat tagcttctaa
3361 aatttggtgt tttcaaacgc tatttacagt acaatctgaa aaaattggat gaacattgta
3421 aatggtagga tacgagttcc aattaatgta ttttttaagt attacactta gcttattaat
3481 tttatatata cattctacct ctgtgggttt aaagacaact cgtacacata ctcaaattat
3541 ttatgaacat tataatgcat attcgtactg tctattaaac atttttttga cattaaaaat
3601 ttcgtttact aacgtagaaa gcgctgataa ttgaggctac tattttaata tctagaaaca
3661 aagattcatt aattaattaa gtaagctcca ggatacttgt atatttcgtt aaaaaagcgc
3721 aatttcaaca attcctatga acatccatga tttcatcaca aaatcaaaca tatcaggatt
3781 taacgtgaat tatttgcttt cttgtgattt tcatctgatt tctggtcatt tacgttactg
3841 taaacaagct ttttaaaaaa aggatttttt acgcaatatt atcgatatcc caatctgtag
3901 tataaaaagt gcaacagtgt tgagtttccc gaaaccaggc atttttgcgg tagggtctga
3961 aactttcacg aaaccatttc acaacaaaac ttcttcaaca acatgtgtgc aaagaaaatc
4021 gtcgtcttac caggagacca tattggccct gaaattgttg cttctgcctt ggaggttttg
4081 aaagtcgttg agaagaagcg acctgagtta aaactcgagt ttgaagaaca caagattgga
4141 ggtgcctcta ttgatgccta tggaacccct ttgactgacg agactgtgaa ggcttgtttg
4201 gaagctgacg gtgttctttt gggtgccgtt ggtggtcctg aatggaccaa ccccaattgt
4261 cgtcctgagc aaggtttatt gaagcttcgt aagagtatgg gtgtttgggc caaccttcga
4321 ccttgcaact ttgccagcaa gtctttagtc aagtacagcc ctttgaagcc tgaaatcgtt
4381 gaaggtgtcg atttttgtgt tgtacgagaa cttactggag gttgttactt tggtgagcgc
4441 actgaggaca acggatcggg ttatgctatg gacacttggc cttacagttt ggaagaagtt
4501 tctcgtattg ctcgtttggc ggccgcgggc ccaacaccaa tgtttataac caagttttat
4561 cttgtttgtc tacatggtat tttacattca tctacataca tctttcattg gctttgtaca
4621 tagttatcat tacaagtcta aaaaaattca ctcttttctt attcaatgtc aatccaagag
4681 aaaagattgt ggtaatgttg taggagcatg tttaataaat tactatagca aattactttt
4741 tattcccaag gtgtttatct ataatagtta atattttagt cgctacataa aattttacca
4801 aagagtactt gtatactaat tctaaatgcc ttctgacata aaacgcctag gaaaacaaac
4861 gcaaacaagg catcgacttt ttcaataacc aaccaaaaaa attttacatt agtctttttt
4921 taatgctgag aaagtctttg ctgatatgcc ttccaaccag cttctctata tctcttggct
4981 tcgacaacag gattacgacc agctccatag actccacgac caacaatgat gatgtcgcta
5041 ccgcagttta caatcacttc ttcaggagta cgatattgct gtcccagccc gtctccttta
5101 acatccaagc cgataccagg ggacatagtt atgtagtcgc tttgaaggtt aggaaatcga
5161 cgaccagcta taaagccaaa gcaaaaatcg gtatgcttct caaaccattc taaggttttc
5221 tctgtgtagg aaccagtagc caaagagcct ttggaagaca tttcagccaa aagcaagaga
5281 ccacgtccca aaggtaaacc aacttctttg aggccttgta taataccctc gcctggcact
5341 gtatggcaat ttgtgatatg agcccaagaa gcaattttgt acacaccaga tgcatattgt
5401 agcttgacgg tatttccaat gtctgcgaat ttgcgatcct caaagataag aaaacgatgc
5461 tttttaccta aggccaccag tttttctacc atatcctggt cgaaatcctc gacaacgtca
5521 atatgtgtct tgataacaca gacatagggt ccaattttat ctaccaattc taagatttcg
5581 gatttcttcg tcaaatcgac cgcgactgac aagttgcttt gcttttcttc catcaaagcc
5641 aacaattcct tggcaatggg atttttcatc ccctcagctc tagctgaata gctttgaaat
5701 actctagcat ccataacttt gcttttaaac ctttaatttc gatccaagca aaaaagaggt
5761 tcttggtagg acaatacggt aagaaaacac gacatgtgca gagatgccga cgaagcatag
5821 ttaaactggg atggtaaaat caattaagaa tttataaaga caaaattgta taagtctcta
5881 aaacatctta attatacctc acagaactat ctaaaatata ttcacaaagt gcaaacatta
5941 tcatgaaaaa gaaccatttt aatttaaagc aagggcatta aggcttattt acagaatttc
6001 ttacttttgt aaagattata aggctgatta tctttttcac catgccaaaa attacacaag
6061 atagaatgga tgtttgaaat taaacgtgag tatacaaaca aatacactag gtaaatcgaa
6121 acattttttt ctccattaag taacaaattc ctatttagag aaagaatgct gagtagatta
6181 aataatctat acaaactttt ttaacacaaa tgcatacata tagccagtgg gatttgtagc
6241 taagcttccg aattcgagtc taactcctta accactcgga catagtgact tatctgacac
6301 tcattgtaaa ttaatatata tataggcatt ttgtttagtt aaaggtactt aagtaattag
6361 tataaacgaa ccaattttat aatcaggaag ttaagtgaat ggtagcacat gtcgtaaaaa
6421 ttgtgaattt ttattgaata atattttaaa tacaagcctt tctagactag gtatactcat
6481 aaacatatat gagcaaaagg atagaggaga ttacattgca tcttctacaa atttatttat
6541 tgccctttac tgaaaaatta aataatgagt actaaatgat aaaaagcgct cagtacaaga
6601 aagattgcaa aaatattgca ttcttcatga attaaagttg catataaggc atattgaaag
6661 taatagtact aaaacagcag ttagcgaaaa ttaatagaat tatattcgca agacaattgt
6721 gacattaaat taaaaaattg tagaattttt actatcctct ttaacgccat gagcctttat
6781 aaaaaggtta aattagtttt aacattcttt ttttgagtaa gagtttacaa tttatcaaaa
6841 cctgtgttat tatattcatt agtttcaatt tattagcatc tagagaaaaa tcaattggca
6901 gttacattgt tggaatttat gaagaaaaag aatctacaac ggagaataag ttgctgatcg
6961 ctttccctaa aattgtatat tttgctgagc ttattttgac atttcgttga agttttctct
7021 aattcgcatt cattttaagt aaaacaatga gaaataaaat tacaaaaaat acaaattaaa
7081 atacaatttt tagctataat tatagacgat gcccttgtat cccattctgt ctcgcttgcc
7141 cctacttttt atcttttata taccataatg aacgctgccg ctactaacca taccccgatt
7201 ttacatttcg gactcccaag gacgtacaaa atagaaaact atagaaaaaa ataatcagaa
7261 aatagcatgt catctctttg taaaacgcgt ttgcaagaag aaaggaaaca atggagaaga
7321 gatcatccat ttgtatgtaa attttagtaa acttgaagaa atcactaaca acttctctta
7381 cttagggatt ctatgcaaaa ccttgtaaat catctgatgg aggactcgat ttaatgaatt
7441 ggaaggttgg gcccgcggcc gcttagctga aacttccaac cctcctgctc ccgtcacatt
7501 actcgacaaa gctaatgttt tggcaacttc tcgtctttgg cgcaagactg tagctaagat
7561 ttttaaggag gaatatcctc accttacctt aaaaaaccag ctcattgact ccgcagccat
7621 gcttttggtc aagagccctc gtacacttaa cggtgttgtt ttgactgaca acttgtttgg
7681 tgacattatt tcagatgagg cttctgtcat tcctggtagc ttgggccttt tgccttctgc
7741 ctccctttcc ggtgtggtag gaaaatcaga agaaaaggtt cattgtttgg ttgagcccat
7801 tcacggtagc gctcccgagc tcgaattcac tggccgtcgt tttacaacgt cgtgactggg
7861 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
7921 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
7981 aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
8041 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
8101 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
8161 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
8221 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
8281 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
8341 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggg
8401 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
8461 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
8521 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
8581 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
8641 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
8701 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
8761 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
8821 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
8881 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
8941 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
9001 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
9061 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
9121 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
9181 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
9241 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
9301 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
9361 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
9421 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
9481 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
9541 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
9601 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
9661 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
9721 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
9781 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
9841 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
9901 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
9961 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
10021 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
10081 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
10141 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
10201 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
10261 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
10321 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
10381 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
10441 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
10501 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
10561 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
10621 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
10681 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
10741 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
10801 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
10861 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
10921 ccatgattac gcc
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