LOCUS Exported 6526 bp ds-DNA circular SYN 10-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 18
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6526)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Saturday, September 10, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbioe.com
FEATURES Location/Qualifiers
source 1..6526
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 1..1165
/note="2u ori"
/note="yeast 2u plasmid origin of replication"
promoter 1447..1551
/gene="bla"
/note="AmpR promoter"
CDS 1552..2412
/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"
rep_origin 2583..3171
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
terminator 3432..3702
/gene="S. cerevisae PGK1"
/note="PGK1 terminator"
/note="transcription terminator for 3-phosphoglycerate
kinase gene"
CDS complement(3828..3848)
/codon_start=1
/product="nuclear localization signal of SV40 large T
antigen"
/note="SV40 NLS"
/translation="PKKKRKV"
CDS complement(3870..3896)
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/note="HA"
/translation="YPYDVPDYA"
promoter complement(3918..4268)
/gene="S. cerevisiae MET17"
/gene_synonym="MET15; MET25"
/note="MET17 promoter"
/note="expression is repressed by methionine"
promoter 4498..4528
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 4536..4552
/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)."
primer_bind 4560..4576
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
promoter 4590..4691
/gene="S. cerevisiae TRP1"
/note="TRP1 promoter"
CDS 4692..5366
/codon_start=1
/gene="S. cerevisiae TRP1"
/product="phosphoribosylanthranilate isomerase, required
for tryptophan biosynthesis"
/note="TRP1"
/note="yeast auxotrophic marker"
/translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
KK"
terminator 5419..5606
/gene="S. cerevisiae ADH1"
/note="ADH1 terminator"
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
CDS complement(5653..6093)
/codon_start=1
/gene="Saccharomyces cerevisiae GAL4 (truncated)"
/product="DNA binding domain of the GAL4 transcriptional
activator"
/note="GAL4 DNA binding domain"
/translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
promoter complement(6121..6517)
/gene="S. cerevisiae ADH1"
/note="ADH1 promoter"
/note="promoter for alcohol dehydrogenase 1"
ORIGIN
1 aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc gctaattttt
61 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa agcgctattt
121 taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa
181 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc
241 tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc
301 gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg ctctataatg
361 cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg
421 tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta
481 gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat
541 gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag
601 aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca
661 ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa
721 gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg
781 agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga
841 tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct cgttacagtc
901 cggtgcgttt ttggtttttt gaaagtgcgt cttcagagcg cttttggttt tcaaaagcgc
961 tctgaagttc ctatactttc tagctagaga ataggaactt cggaatagga acttcaaagc
1021 gtttccgaaa acgagcgctt ccgaaaatgc aacgcgagct gcgcacatac agctcactgt
1081 tcacgtcgca cctatatctg cgtgttgcct gtatatatat atacatgaga agaacggcat
1141 agtgcgtgtt tatgcttaaa tgcgttatgg tgcactctca gtacaatctg ctctgatgcc
1201 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
1261 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
1321 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
1381 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
1441 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
1501 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
1561 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
1621 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
1681 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
1741 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
1801 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
1861 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
1921 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
1981 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
2041 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
2101 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
2161 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
2221 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
2281 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
2341 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
2401 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
2461 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
2521 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
2581 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
2641 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
2701 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
2761 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
2821 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
2881 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
2941 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
3001 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
3061 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
3121 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
3181 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
3241 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
3301 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
3361 atacgcaaac gcctctcccc gcgcgttggc cgattcatta atgcaggaag atccgaggcc
3421 tagctttaac gaacgcagaa ttttcgagtt attaaactta aaatacgctg aacccgaaca
3481 tagaaatatc gaatgggaaa aaaaaactgc ataaaggcat taaaagagga gcgaattttt
3541 ttttaataaa aatcttaata atcattaaaa gataaataat agtctatata tacgtatata
3601 aataaaaaat attcaaaaaa taaaataaac tattatttta gcgtaaagga tggggaaaga
3661 gaaaagaaaa aaattgatct atcgatttca attcaattca atttatttct tttcggataa
3721 gaaagcaaca cctggcaatt ccttaccttc caataattcc aaagaagcac caccaccagt
3781 agagacatgg gagatcagcc cgaagatctt cgggctaatg cggccgccac ctttctcttc
3841 ttcttaggac tagaaggtcc tcccaggctg gcatagtcag gcacgtcata aggatagcta
3901 gaagccatat ggcccattgt atggatgggg gtaatagaat tgtatctatg tatctgacga
3961 ccctgtatta cacgaaggaa atagtagatg aaaagacaag agagcaagaa aaaggaaaaa
4021 cttcctttct aacagacgct ttacttaacc ttatatatat cttatttttt tcatcgagcg
4081 tgttcaattg gacaaggtgc caccttttcg acacttcggt tattatgtta cacagttttc
4141 atgaggatgg cgccttgact aacttaatta gtcatttgcc aaccaccaca gttccccaat
4201 atcgacagct tcacgtgcca tttgccattt tgcgatcatg tgacctcaag agaaaaagca
4261 aaaaataatg agagctaagg gattcgaacc cttgcatccg aagatatcag agattttaga
4321 ggttaaatcc acctaaatct gacgccttaa accactcggc caaactccca ttgtatgttg
4381 gaagaattag aagctaggcc tcggatcttc ctggcacgac aggtttcccg actggaaagc
4441 gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact cattaggcac cccaggcttt
4501 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
4561 aggaaacagc tatgacatga ttacgaatta attcggtcga aaaaagaaaa ggagagggcc
4621 aagagggagg gcattggtga ctattgagca cgtgagtata cgtgattaag cacacaaagg
4681 cagcttggag tatgtctgtt attaatttca caggtagttc tggtccattg gtgaaagttt
4741 gcggcttgca gagcacagag gccgcagaat gtgctctaga ttccgatgct gacttgctgg
4801 gtattatatg tgtgcccaat agaaagagaa caattgaccc ggttattgca aggaaaattt
4861 caagtcttgt aaaagcatat aaaaatagtt caggcactcc gaaatacttg gttggcgtgt
4921 ttcgtaatca acctaaggag gatgttttgg ctctggtcaa tgattacggc attgatatcg
4981 tccaactgca tggagatgag tcgtggcaag aataccaaga gttcctcggt ttgccagtta
5041 ttaaaagact cgtatttcca aaagactgca acatactact cagtgcagct tcacagaaac
5101 ctcattcgtt tattcccttg tttgattcag aagcaggtgg gacaggtgaa cttttggatt
5161 ggaactcgat ttctgactgg gttggaaggc aagagagccc cgaaagctta cattttatgt
5221 tagctggtgg actgacgcca gaaaatgttg gtgatgcgct tagattaaat ggcgttattg
5281 gtgttgatgt aagcggaggt gtggagacaa atggtgtaaa agactctaac aaaatagcaa
5341 atttcgtcaa aaatgctaag aaataggtta ttactgagta gtatttattt aagtattgtt
5401 tgtgcacttg ccggcatgcc ggtagaggtg tggtcaataa gagcgacctc atgctatacc
5461 tgagaaagca acctgaccta caggaaagag ttactcaaga acaagaattt tcgttttaaa
5521 acctaagagt cactttaaaa tttgtataca cttatttttt ttataactta tttaataata
5581 aaaatcataa atcataagaa attcgcccgg aattagcttg gctgcaggtc gacggatccc
5641 cgggaattcc ggcgatacag tcaactgtct ttgacctttg ttactactct cttccgatga
5701 tgatgtcgca cttattctat gctgtctcaa tgttagaggc atatcagtct ccactgaagc
5761 caatctatct gtgacggcat ctttattcac attatcttgt acaaataatc ctgttaacaa
5821 tgcttttata tcctgtaaag aatccatttt caaaatcatg tcaaggtctt ctcgaggaaa
5881 aatcagtaga aatagctgtt ccagtctttc tagccttgat tccacttctg tcagatgtgc
5941 cctagtcagc ggagaccttt tggttttggg agagtagcga cactcccagt tgttcttcag
6001 acacttggcg cacttcggtt tttctttgga gcacttgagc tttttaagtc ggcaaatatc
6061 gcatgcttgt tcgatagaag acagtagctt catctttcag gaggcttgct tcaagcttgg
6121 agttgattgt atgcttggta tagcttgaaa tattgtgcag aaaaagaaac aaggaagaaa
6181 gggaacgaga acaatgacga ggaaacaaaa gattaataat tgcaggtcta tttatacttg
6241 atagcaagac agcaaacttt tttttatttc aaattcaagt aactggaagg aaggccgtat
6301 accgttgctc attagagagt agtgtgcgtg aatgaaggaa ggaaaaagtt tcgtgtgctt
6361 cgagataccc ctcatcagct ctggaacaac gacatctgtt ggtgctgtct ttgtcgttaa
6421 ttttttcctt tagtgtcttc catcattttt ttgtcattgc ggatatggtg agacaacaac
6481 gggggagaga gaaaagaaaa aaaaagaaaa gaagttgcat gcaagc
//