LOCUS Exported 6412 bp ds-DNA circular SYN 12-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 9
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6412)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Monday, September 12, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..6412
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 201..416
/gene="S. cerevisiae URA3"
/note="URA3 promoter"
CDS 417..1220
/codon_start=1
/gene="S. cerevisiae URA3"
/product="orotidine-5'-phosphate decarboxylase, required
for uracil biosynthesis"
/note="URA3"
/note="yeast auxotrophic marker, counterselectable with
5-fluoroorotic acid (5-FOA)"
/translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
rep_origin complement(1351..1806)
/direction=LEFT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 1951..1967
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter 1977..1995
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
terminator 2014..2261
/gene="S. cerevisiae CYC1"
/note="CYC1 terminator"
/note="transcription terminator for CYC1"
primer_bind 2264..2280
/note="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2314..2330)
/note="SK primer"
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(2345..2786)
/gene="S. cerevisiae GAL1"
/note="GAL1 promoter"
/note="inducible promoter, regulated by Gal4"
protein_bind 2669..2786
/bound_moiety="Gal4"
/note="UAS"
/note="upstream activating sequence mediating
Gal4-dependent induction"
promoter complement(2810..2828)
/note="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(2849..2865)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2873..2889
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/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)."
promoter complement(2897..2927)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
rep_origin complement(3251..3839)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4010..4870)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(4871..4975)
/gene="bla"
/note="AmpR promoter"
rep_origin 5002..6344
/note="2u ori"
/note="yeast 2u plasmid origin of replication"
protein_bind complement(5965..6012)
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FRT"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
ORIGIN
1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
181 accataccac agcttttcaa ttcaattcat catttttttt ttattctttt ttttgatttc
241 ggtttctttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
301 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
361 cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaatcatgt
421 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
481 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
541 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
601 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
661 ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt aatacagtca
721 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
781 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
841 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
901 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
961 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
1021 ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga accgtggatg
1081 atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
1141 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
1201 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac
1261 aaattagagc ttcaatttaa ttatatcagt tattacccta tgcggtgtga aataccgcac
1321 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat
1381 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
1441 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
1501 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
1561 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
1621 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
1681 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
1741 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
1801 gcgcgtcgcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
1861 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
1921 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat
1981 acgactcact atagggcgaa ttgggtaccg gccgcaaatt aaagccttcg agcgtcccaa
2041 aaccttctca agcaaggttt tcagtataat gttacatgcg tacacgcgtc tgtacagaaa
2101 aaaaagaaaa atttgaaata taaataacgt tcttaatact aacataacta taaaaaaata
2161 aatagggacc tagacttcag gttgtctaac tccttccttt tcggttagag cggatgtggg
2221 gggagggcgt gaatgtaagc gtgacataac taattacatg actcgaggtc gacggtatcg
2281 ataagcttga tatcgaattc ctgcagcccg ggggatccac tagttctaga atccggggtt
2341 ttttctcctt gacgttaaag tatagaggta tattaacaat tttttgttga tacttttatt
2401 acatttgaat aagaagtaat acaaaccgaa aatgttgaaa gtattagtta aagtggttat
2461 gcagtttttg catttatata tctgttaata gatcaaaaat catcgcttcg ctgattaatt
2521 accccagaaa taaggctaaa aaactaatcg cattatcatc ctatggttgt taatttgatt
2581 cgttcatttg aaggtttgtg gggccaggtt actgccaatt tttcctcttc ataaccataa
2641 aagctagtat tgtagaatct ttattgttcg gagcagtgcg gcgcgaggca catctgcgtt
2701 tcaggaacgc gaccggtgaa gacgaggacg cacggaggag agtcttcctt cggagggctg
2761 tcacccgctc ggcggcttct aatccgtact agagctccag cttttgttcc ctttagtgag
2821 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
2881 cgctcacaat tccacacaac ataggagccg gaagcataaa gtgtaaagcc tggggtgcct
2941 aatgagtgag gtaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
3001 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
3061 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
3121 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
3181 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
3241 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
3301 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
3361 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
3421 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
3481 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
3541 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
3601 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
3661 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
3721 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
3781 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
3841 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
3901 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
3961 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
4021 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
4081 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
4141 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
4201 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
4261 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
4321 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
4381 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
4441 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
4501 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
4561 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
4621 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
4681 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
4741 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
4801 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
4861 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
4921 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
4981 ttccccgaaa agtgccacct gaacgaagca tctgtgcttc attttgtaga acaaaaatgc
5041 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
5101 tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt gtaaaacaaa
5161 aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac
5221 agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct
5281 acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc
5341 tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag
5401 gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact
5461 tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc
5521 ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt
5581 tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac
5641 tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt
5701 actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg
5761 agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca
5821 cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaata
5881 ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg tcttcagagc
5941 gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagagaata ggaacttcgg
6001 aataggaact tcaaagcgtt tccgaaaacg agcgcttccg aaaatgcaac gcgagctgcg
6061 cacatacagc tcactgttca cgtcgcacct atatctgcgt gttgcctgta tatatatata
6121 catgagaaga acggcatagt gcgtgtttat gcttaaatgc gtacttatat gcgtctattt
6181 atgtaggatg aaaggtagtc tagtacctcc tgtgatatta tcccattcca tgcggggtat
6241 cgtatgcttc cttcagcact accctttagc tgttctatat gctgccactc ctcaattgga
6301 ttagtctcat ccttcaatgc tatcatttcc tttgatattg gatcatacta agaaaccatt
6361 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tc
//