LOCUS Exported 6038 bp ds-DNA circular SYN 10-NOV-2017
DEFINITION synthetic circular DNA
REFERENCE 1 (bases 1 to 6038)
FEATURES Location/Qualifiers
source 1..6038
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 2..443
/gene="S. cerevisiae GAL1"
/label=GAL1 promoter
/note="inducible promoter, regulated by Gal4"
protein_bind 2..119
/label=UAS
/bound_moiety="Gal4"
/note="upstream activating sequence mediating
Gal4-dependent induction"
promoter 475..493
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 522..539
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 543..575
/codon_start=1
/product="leader peptide from bacteriophage T7 gene 10"
/label=T7 tag (gene 10 leader)
/note="promotes efficient translation in E. coli"
/translation="MASMTGGQQMG"
CDS 579..602
/codon_start=1
/product="Xpress(TM) epitope tag, including an enterokinase
recognition and cleavage site"
/label=Xpress(TM) tag
/translation="DLYDDDDK"
CDS 682..723
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
CDS 733..750
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
terminator 784..1031
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
rep_origin complement(1279..1867)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2038..2898)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCDTLLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
CDS complement(2994..3797)
/codon_start=1
/gene="S. cerevisiae URA3"
/product="orotidine-5'-phosphate decarboxylase, required
for uracil biosynthesis"
/label=URA3
/note="yeast auxotrophic marker, counterselectable with
5-fluoroorotic acid (5-FOA)"
/translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
promoter complement(3798..4013)
/gene="S. cerevisiae URA3"
/label=URA3 promoter
rep_origin 4617..5497
/label=2u ori
/note="yeast 2u plasmid origin of replication"
rep_origin complement(5566..6021)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
ORIGIN
1 acggattaga agccgccgag cgggtgacag ccctccgaag gaagactctc ctccgtgcgt
61 cctcgtcttc accggtcgcg ttcctgaaac gcagatgtgc ctcgcgccgc actgctccga
121 acaataaaga ttctacaata ctagctttta tggttatgaa gaggaaaaat tggcagtaac
181 ctggccccac aaaccttcaa atgaacgaat caaattaaca accataggat gataatgcga
241 ttagtttttt agccttattt ctggggtaat taatcagcga agcgatgatt tttgatctat
301 taacagatat ataaatgcaa aaactgcata accactttaa ctaatacttt caacattttc
361 ggtttgtatt acttcttatt caaatgtaat aaaagtatca acaaaaaatt gttaatatac
421 ctctatactt taacgtcaag gagaaaaaac cccggatcgg actactagca gctgtaatac
481 gactcactat agggaatatt aagcttacca tggggggttc tcatcatcat catcatcatg
541 gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac gatgacgata
601 aggtacctag gatccagtgt ggtggaattc tgcagatatc cagcacagtg gcggccgctc
661 gagtctagag ggcccttcga aggtaagcct atccctaacc ctctcctcgg tctcgattct
721 acgcgtaccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc ctagagggcc
781 gcatcatgta attagttatg tcacgcttac attcacgccc tccccccaca tccgctctaa
841 ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt tttatagtta
901 tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg tacagacgcg
961 tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg
1021 ctttaatttg caagctgcgg ccctgcatta atgaatcggc caacgcgcgg ggagaggcgg
1081 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
1141 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
1201 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aagcccagga accgtaaaaa
1261 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
1321 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
1381 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
1441 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
1501 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
1561 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
1621 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
1681 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
1741 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
1801 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
1861 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
1921 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
1981 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
2041 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
2101 tgcctgactc cccgtcgtgt agataactac gatacgggag cgcttaccat ctggccccag
2161 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
2221 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
2281 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
2341 tgttggcatt gctacaggca tcgtggtgtc actctcgtcg tttggtatgg cttcattcag
2401 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
2461 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
2521 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
2581 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
2641 ttgcccggcg tcaatacggg ataatagtgt atcacatagc agaactttaa aagtgctcat
2701 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
2761 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
2821 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
2881 gaaatgttga atactcatac tcttcctttt tcaatgggta ataactgata taattaaatt
2941 gaagctctaa tttgtgagtt tagtatacat gcatttactt ataatacagt tttttagttt
3001 tgctggccgc atcttctcaa atatgcttcc cagcctgctt ttctgtaacg ttcaccctct
3061 accttagcat cccttccctt tgcaaatagt cctcttccaa caataataat gtcagatcct
3121 gtagagacca catcatccac ggttctatac tgttgaccca atgcgtctcc cttgtcatct
3181 aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt catctcttcc acccatgtct
3241 ctttgagcaa taaagccgat aacaaaatct ttgtcgctct tcgcaatgtc aacagtaccc
3301 ttagtatatt ctccagtaga tagggagccc ttgcatgaca attctgctaa catcaaaagg
3361 cctctaggtt cctttgttac ttcttctgcc gcctgcttca aaccgctaac aatacctggg
3421 cccaccacac cgtgtgcatt cgtaatgtct gcccattctg ctattctgta tacacccgca
3481 gagtactgca atttgactgt attaccaatg tcagcaaatt ttctgtcttc gaagagtaaa
3541 aaattgtact tggcggataa tgcctttagc ggcttaactg tgccctccat ggaaaaatca
3601 gtcaagatat ccacatgtgt ttttagtaaa caaattttgg gacctaatgc ttcaactaac
3661 tccagtaatt ccttggtggt acgaacatcc aatgaagcac acaagtttgt ttgcttttcg
3721 tgcatgatat taaatagctt ggcagcaaca ggactaggat gagtagcagc acgttcctta
3781 tatgtagctt tcgacatgat ttatcttcgt ttcctgcagg tttttgttct gtgcagttgg
3841 gttaagaata ctgggcaatt tcatgtttct tcaacactac atatgcgtat atataccaat
3901 ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg agattaccga atcaaaaaaa
3961 tttcaaagaa accgaaatca aaaaaaagaa taaaaaaaaa atgatgaatt gaattgaaaa
4021 gctagcttat cgatgataag ctgtcaaaga tgagaattaa ttccacggac tatagactat
4081 actagatact ccgtctactg tacgatacac ttccgctcag gtccttgtcc tttaacgagg
4141 ccttaccact cttttgttac tctattgatc cagctcagca aaggcagtgt gatctaagat
4201 tctatcttcg cgatgtagta aaactagcta gaccgagaaa gagactagaa atgcaaaagg
4261 cacttctaca atggctgcca tcattattat ccgatgtgac gctgcagctt ctcaatgata
4321 ttcgaatacg ctttgaggag atacagccta atatccgaca aactgtttta cagatttacg
4381 atcgtacttg ttacccatca ttgaattttg aacatccgaa cctgggagtt ttccctgaaa
4441 cagatagtat atttgaacct gtataataat atatagtcta gcgctttacg gaagacaatg
4501 tatgtatttc ggttcctgga gaaactattg catctattgc ataggtaatc ttgcacgtcg
4561 catccccggt tcattttctg cgtttccatc ttgcacttca atagcatatc tttgttaacg
4621 aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa
4681 caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg ctattttacc
4741 aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc aacgcgacga gagcgctaat
4801 ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg cgagagcgct
4861 attttaccaa caaagaatct atacttcttt tttgttctac aaaaatgcat cccgagagcg
4921 ctatttttct aacaaagcat cttagattac tttttttctc ctttgtgcgc tctataatgc
4981 agtctcttga taactttttg cactgtaggt ccgttaaggt tagaagaagg ctactttggt
5041 gtctattttc tcttccataa aaaaagcctg actccacttc ccgcgtttac tgattactag
5101 cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc gattatattc tataccgatg
5161 tggattgcgc atactttgtg aacagaaagt gatagcgttg atgattcttc attggtcaga
5221 aaattatgaa cggtttcttc tattttgtct ctatatacta cgtataggaa atgtttacat
5281 tttcgtattg ttttcgattc actctatgaa tagttcttac tacaattttt ttgtctaaag
5341 agtaatacta gagataaaca taaaaaatgt agaggtcgag tttagatgca agttcaagga
5401 gcgaaaggtg gatgggtagg ttatataggg atatagcaca gagatatata gcaaagagat
5461 acttttgagc aatgtttgtg gaagcggtat tcgcaatggg aagctccacc ccggttgata
5521 atcagaaaag ccccaaaaac aggaagattg tataagcaaa tatttaaatt gtaaacgtta
5581 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aacgaatagc
5641 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
5701 ttccagtttc caacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
5761 aaagggtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
5821 ggtcgaggtg ccgtaaagca gtaaatcgga agggtaaacg gatgccccca tttagagctt
5881 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcggggg
5941 ctagggcggt gggaagtgta ggggtcacgc tgggcgtaac caccacaccc gccgcgctta
6001 atggggcgct acagggcgcg tggggatgat ccactagt
//