LOCUS Exported 8058 bp ds-DNA circular SYN 10-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 22
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8058)
AUTHORS .
TITLE Direct Submission
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..8058
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 4..1168
/note="2u ori"
/note="yeast 2u plasmid origin of replication"
promoter 1450..1554
/gene="bla"
/note="AmpR promoter"
CDS 1555..2415
/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 2586..3174
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 3414..3447
/bound_moiety="Cre recombinase"
/note="loxP"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
protein_bind 3525..3558
/bound_moiety="Cre recombinase"
/note="loxP"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
protein_bind 3613..3634
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3649..3679
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 3687..3703
/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 3711..3727
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
promoter 3769..4174
/gene="S. cerevisiae LEU2"
/note="LEU2 promoter"
CDS 4175..5269
/codon_start=1
/gene="S. cerevisiae LEU2"
/product="3-isopropylmalate dehydrogenase, required for
leucine biosynthesis"
/note="LEU2"
/note="yeast auxotrophic marker"
/translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
IGGAAIEATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
DLGGSNSTTEVGDAVAEEVKKILA"
terminator 5386..5573
/gene="S. cerevisiae ADH1"
/note="ADH1 terminator"
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
CDS complement(6092..6118)
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/note="HA"
/translation="YPYDVPDYA"
promoter complement(6137..6155)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS complement(6161..6502)
/codon_start=1
/gene="Saccharomyces cerevisiae GAL4 (truncated)"
/product="activation domain of the GAL4 transcriptional
activator"
/note="GAL4 activation domain"
/translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
EDTPPNPKKE"
CDS complement(6518..6538)
/codon_start=1
/product="nuclear localization signal of SV40 large T
antigen"
/note="SV40 NLS"
/translation="PKKKRKV"
promoter complement(6584..7288)
/gene="S. cerevisiae ADH1"
/note="ADH1 promoter"
/note="promoter for alcohol dehydrogenase 1"
ORIGIN
1 agcaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag agcgctaatt
61 tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc gaaagcgcta
121 ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc
181 taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgagag
241 cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat gcatcccgag
301 agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt gcgctctata
361 atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag aaggctactt
421 tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt ttactgatta
481 ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat attctatacc
541 gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt cttcattggt
601 cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata ggaaatgttt
661 acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat ttttttgtct
721 aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga tgcaagttca
781 aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata tatagcaaag
841 agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tattttagta gctcgttaca
901 gtccggtgcg tttttggttt tttgaaagtg cgtcttcaga gcgcttttgg ttttcaaaag
961 cgctctgaag ttcctatact ttctagctag agaataggaa cttcggaata ggaacttcaa
1021 agcgtttccg aaaacgagcg cttccgaaaa tgcaacgcga gctgcgcaca tacagctcac
1081 tgttcacgtc gcacctatat ctgcgtgttg cctgtatata tatatacatg agaagaacgg
1141 catagtgcgt gtttatgctt aaatgcgtta tggtgcactc tcagtacaat ctgctctgat
1201 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
1261 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
1321 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
1381 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
1441 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
1501 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
1561 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
1621 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
1681 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
1741 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
1801 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
1861 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
1921 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
1981 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
2041 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
2101 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
2161 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
2221 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
2281 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
2341 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
2401 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
2461 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
2521 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
2581 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
2641 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
2701 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
2761 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
2821 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
2881 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
2941 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
3001 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
3061 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
3121 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
3181 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
3241 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
3301 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
3361 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctgataactt
3421 cgtatagcat acattatacg aagttatatt aagggttccg gatcgcggcc gctcgacctg
3481 cagccaagct agcttggctg gacgtaaact cctcttcaga cctaataact tcgtatagca
3541 tacattatac gaagttatca gctggcacga caggtttccc gactggaaag cgggcagtga
3601 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
3661 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
3721 ctatgacatg attacgaatt gatcctctag ctagagtcga gatcccccaa ctgtgggaat
3781 actcaggtat cgtaagatgc aagagttcga atctcttagc aaccattatt tttttcctca
3841 acataacgag aacacacagg ggcgctatcg cacagaatca aattcgatga ctggaaattt
3901 tttgttaatt tcagaggtcg cctgacgcat ataccttttt caactgaaaa attgggagaa
3961 aaaggaaagg tgagagcgcc ggaaccggct tttcatatag aatagagaag cgttcatgac
4021 taaatgcttg catcacaata cttgaagttg acaatattat ttaaggacct attgtttttt
4081 ccaataggtg gttagcaatc gtcttacttt ctaacttttc ttacctttta catttcagca
4141 atatatatat atatttcaag gatataccat tctaatgtct gcccctaaga agatcgtcgt
4201 tttgccaggt gaccacgttg gtcaagaaat cacagccgaa gccattaagg ttcttaaagc
4261 tatttctgat gttcgttcca atgtcaagtt cgatttcgaa aatcatttaa ttggtggtgc
4321 tgctatcgaa gctacaggtg tcccacttcc agatgaggcg ctggaagcct ccaagaaggc
4381 tgatgccgtt ttgttaggtg ctgtgggtgg tcctaaatgg ggtacaggta gtgttagacc
4441 tgaacaaggt ttactaaaaa tccgtaaaga acttcaattg tacgccaact taagaccatg
4501 taactttgca tccgactctc ttttagactt atctccaatc aagccacaat ttgctaaagg
4561 tactgacttc gttgttgtca gagaattagt gggaggtatt tactttggta agagaaagga
4621 agacgatggt gatggtgtcg cttgggatag tgaacaatac accgttccag aagtgcaaag
4681 aatcacaaga atggccgctt tcatggccct acaacatgag ccaccattgc ctatttggtc
4741 cttggataaa gctaatgttt tggcctcttc aagattatgg agaaaaactg tggaggaaac
4801 catcaagaac gaatttccta cattgaaggt tcaacatcaa ttgattgatt ctgccgccat
4861 gatcctagtt aagaacccaa cccacctaaa tggtattata atcaccagca acatgtttgg
4921 tgatatcatc tccgatgaag cctccgttat cccaggttcc ttgggtttgt tgccatctgc
4981 gtccttggcc tctttgccag acaagaacac cgcatttggt ttgtacgaac catgccacgg
5041 ttctgctcca gatttgccaa agaataaggt caaccctatc gccactatct tgtctgctgc
5101 aatgatgttg aaattgtcat tgaacttgcc tgaagaaggt aaggccattg aagatgcagt
5161 taaaaaggtt ttggatgcag gtatcagaac tggtgattta ggtggttcca acagtaccac
5221 cgaagtcggt gatgctgtcg ccgaagaagt taagaaaatc cttgcttaaa aagattctct
5281 ttttttatga tatttgtaca taaactttat aaatgaaatt cataatagaa acgacacgaa
5341 attacaaaat ggaatatgtt catagggtag gggaatttcg accggccggt agaggtgtgg
5401 tcaataagag cgacctcatg ctatacctga gaaagcaacc tgacctacag gaaagagtta
5461 ctcaagaata agaattttcg ttttaaaacc taagagtcac tttaaaattt gtatacactt
5521 atttttttta taacttattt aataataaaa atcataaatc ataagaaatt cgcttattta
5581 gaagtgtcaa caacgtatct accaacgatt tgaccctttt ccatcttttc gtaaatttct
5641 ggcaaggtag acaagccgac aaccttgatt ggagacttga ccaaacctct ggcgaagaag
5701 tccaaagctt ctgaataagc cctcgtaata tattttcatg aagaatttag gtccaaaaaa
5761 aagatgggca ttaattctag tcatttaaaa aattctatag atcagaggtt acatggccaa
5821 gattgaaact tagaggagta tagttacata aaagaaggca aaacgatgta taaatgaaag
5881 aaattgagat ggtgcacgat gcacagttga agtgaacttg cggggttttt cagtatctac
5941 gattcatctg cagctcgagc tcgatggatc ccgtatcgat gcccaccctc tagaggccga
6001 ggcggccgac atgttttttc ccgggccata atggccactc tgcgttgata ccactgcttg
6061 ggtggaattc actggcctcc atggccatat gagcgtaatc tggtacgtcg tatgggtact
6121 ccatggcggc gctcgcccta tagtgagtcg tattaaagat ctcttttttt gggtttggtg
6181 gggtatcttc atcatcgaat agatagttat atacatcatc cattgtagtg gtattaaaca
6241 tccctgtagt gattccaaac gcgttatacg cagtttggtc cgtccaacca ggtgacagtg
6301 gttttgaatt attaccatca tcaattttac tagccgtgat ttcattattc atgaagttat
6361 catgaacgtt agaggaggca attggttgtg aaagcgcttg agaatttgtt tgagttgtta
6421 tgaggttcgg accgttgcta ctgttagtga aagtgaagga caatgagcta tcagcaatat
6481 tcccactttg attaaaattg gcggcggtac ccaattcgac ctttctcttc ttttttggag
6541 gctcgggaat taattccgct ttatccatct ttgcaaagct tggagttgat tgtatgcttg
6601 gtatagcttg aaatattgtg cagaaaaaga aacaaggaag aaagggaacg agaacaatga
6661 cgaggaaaca aaagattaat aattgcaggt ctatttatac ttgatagcaa gacagcaaac
6721 ttttttttat ttcaaattca agtaactgga aggaaggccg tataccgttg ctcattagag
6781 agtagtgtgc gtgaatgaag gaaggaaaaa gtttcgtgtg cttcgagata cccctcatca
6841 gctctggaac aacgacatct gttggtgctg tctttgtcgt taattttttc ctttagtgtc
6901 ttccatcatt tttttgtcat tgcggatatg gtgagacaac aacgggggag agagaaaaga
6961 aaaaaaaaga aaagaagttg catgcgccta ttattacttc aatagatggc aaatggaaaa
7021 agggtagtga aacttcgata tgatgatggc tatcaagtct agggctacag tattagttcg
7081 ttatgtacca ccatcaatga ggcagtgtaa ttggtgtagt cttgtttagc ccattatgtc
7141 ttgtctggta tctgttctat tgtatatctc ccctccgcca cctacatgtt agggagacca
7201 acgaaggtat tataggaatc ccgatgtatg ggtttggttg ccagaaaaga ggaagtccat
7261 attgtacacc cggaaacaac aaaaggatat ccgaaatatt ccacggttta gaaaaaaatc
7321 ggaaaagagc gcggaggggt gttacccccc ttctctacta gcattggact ttaattaata
7381 tatgtgcata ggagaagtgt aaagttccct tccatattgt aacataataa agtgcacacc
7441 caaatgaatt gaaagcgtac tcaaacagac aaccatttcc agtgttgtat gtacctgtct
7501 atttatactg gtagcaaccc tattgctgtt tcctcttcaa agtactctag cggttatgcg
7561 cgtctcacct tcaaggtcat ggtcgctcta ttgttcgcac caccggcaaa ctcgcgtctc
7621 gcaagtcttg gctcattctt ctagtatact catgttgcaa atgcactcag gttctttcgg
7681 caacttaaat aatgacacca gttgtcgtgg tcgtcatcat cgcaacccca accggcattc
7741 ttattgcttc tccaatctcg ccccttagcg cagggtaaac cttggaaaat gcaggcgcaa
7801 aaaactccgc cgggcacagc ctcacgccca gcgttatcgc cgggccggca agagcgcggg
7861 tccgccacag agtcagcatg attgtgcaat tgcgtaaact cgttttttcg gcgccgcaaa
7921 gccaaataca tcatatcaac acttttcact ttatttttcg ttcgaccctt atatttgtct
7981 tttgccttca tgctccttga tttcctattt catttaccat catttcttcg atcccggatc
8041 tcgacctgca ggcatgca
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