LOCUS Exported 7304 bp ds-DNA circular SYN 10-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 19
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7304)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Saturday, September 10, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..7304
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 30..734
/gene="S. cerevisiae ADH1"
/note="ADH1 promoter"
/note="promoter for alcohol dehydrogenase 1"
CDS 762..1202
/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 1213..1231
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1251..1280
/codon_start=1
/product="Myc (human c-Myc oncogene) epitope tag"
/note="Myc"
/translation="EQKLISEEDL"
terminator 1338..1385
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
terminator 1412..1599
/gene="S. cerevisiae ADH1"
/note="ADH1 terminator"
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
primer_bind complement(1623..1639)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 1647..1663
/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(1671..1701)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
rep_origin complement(2025..2613)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
polyA_signal 2942..2989
/note="HSV TK poly(A) signal"
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
CDS complement(3222..4016)
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/note="NeoR/KanR"
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHDDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
promoter complement(4017..4121)
/gene="bla"
/note="AmpR promoter"
rep_origin 4148..5490
/note="2u ori"
/note="yeast 2u plasmid origin of replication"
protein_bind complement(5111..5158)
/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)."
promoter 5749..6030
/gene="S. cerevisiae TRP1"
/note="TRP1 promoter"
CDS 6031..6705
/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"
rep_origin complement(6808..7263)
/direction=LEFT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
ORIGIN
1 cggtgcgggc ctcttcgcta ttacgccaga tccttttgtt gtttccgggt gtacaatatg
61 gacttcctct tttctggcaa ccaaacccat acatcgggat tcctataata ccttcgttgg
121 tctccctaac atgtaggtgg cggaggggag atatacaata gaacagatac cagacaagac
181 ataatgggct aaacaagact acaccaatta cactgcctca ttgatggtgg tacataacga
241 actaatactg tagccctaga cttgatagcc atcatcatat cgaagtttca ctaccctttt
301 tccatttgcc atctattgaa gtaataatag gcgcatgcaa cttcttttct ttttttttct
361 tttctctctc ccccgttgtt gtctcaccat atccgcaatg acaaaaaaaa tgatggaaga
421 cactaaagga aaaaattaac gacaaagaca gcaccaacag atgtcgttgt tccagagctg
481 atgaggggta tctcgaagca cacgaaactt tttccttcct tcattcacgc acactactct
541 ctaatgagca acggtatacg gccttccttc cagttacttg aatttgaaat aaaaaaagtt
601 tgctgtcttg ctatcaagta taaatagacc tgcaattatt aatcttttgt ttcctcgtca
661 ttgttctcgt tccctttctt ccttgtttct ttttctgcac aatatttcaa gctataccaa
721 gcatacaatc aactccaagc ttgaagcaag cctcctgaaa gatgaagcta ctgtcttcta
781 tcgaacaagc atgcgatatt tgccgactta aaaagctcaa gtgctccaaa gaaaaaccga
841 agtgcgccaa gtgtctgaag aacaactggg agtgtcgcta ctctcccaaa accaaaaggt
901 ctccgctgac tagggcacat ctgacagaag tggaatcaag gctagaaaga ctggaacagc
961 tatttctact gatttttcct cgagaagacc ttgacatgat tttgaaaatg gattctttac
1021 aggatataaa agcattgtta acaggattat ttgtacaaga taatgtgaat aaagatgccg
1081 tcacagatag attggcttca gtggagactg atatgcctct aacattgaga cagcatagaa
1141 taagtgcgac atcatcatcg gaagagagta gtaacaaagg tcaaagacag ttgactgtat
1201 cgccggaatt tgtaatacga ctcactatag ggcgagccgc catcatggag gagcagaagc
1261 tgatctcaga ggaggacctg catatggcca tggaggccga attcccgggg atccgtcgac
1321 ctgcagcggc cgcataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
1381 ttttgcgcgc ttgcagccaa gctaattccg ggcgaatttc ttatgattta tgatttttat
1441 tattaaataa gttataaaaa aaataagtgt atacaaattt taaagtgact cttaggtttt
1501 aaaacgaaaa ttcttattct tgagtaactc tttcctgtag gtcaggttgc tttctcaggt
1561 atagcatgag gtcgctctta ttgaccacac ctctaccggc atgcaagctt ggcgtaatca
1621 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
1681 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgaggtaact cacattaatt
1741 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
1801 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
1861 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
1921 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
1981 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
2041 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
2101 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
2161 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
2221 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
2281 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
2341 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
2401 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
2461 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
2521 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
2581 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
2641 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
2701 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
2761 tatgagtaac ctgaggctat ggcagggcct gccgccccga cgttggctgc gagccctggg
2821 ccttcacccg aacttggggg gtggggtggg gaaaaggaag aaacgcgggc gtattggccc
2881 caatggggtc tcggtggggt atcgacagag tgccagccct gggaccgaac cccgcgttta
2941 tgaacaaacg acccaacacc gtgcgtttta ttctgtcttt ttattgccgt catagcgcgg
3001 gttccttccg gtattgtctc cttccgtgtt tcagttagcc tccccctagg gtgggcgaag
3061 aactccagca tgagatcccc gcgctggagg atcatccagc cggcgtcccg gaaaacgatt
3121 ccgaagccca acctttcata gaaggcggcg gtggaatcga aatctcgtga tggcaggttg
3181 ggcgtcgctt ggtcggttca tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga
3241 aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag
3301 cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc
3361 tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt
3421 tccaccatga tattcggcaa gcaggcatcg tcatgggtca cgacgagatc ctcgccgtcg
3481 ggcatgctcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg
3541 tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga
3601 tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt
3661 gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc
3721 cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca
3781 gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcttgcagt
3841 tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac
3901 agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat
3961 agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatactc
4021 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
4081 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
4141 ccacctgaac gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg cgagagcgct
4201 aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgaaagc
4261 gctattttac caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg caacgcgaga
4321 gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg
4381 agagcgctat tttaccaaca aagaatctat acttcttttt tgttctacaa aaatgcatcc
4441 cgagagcgct atttttctaa caaagcatct tagattactt tttttctcct ttgtgcgctc
4501 tataatgcag tctcttgata actttttgca ctgtaggtcc gttaaggtta gaagaaggct
4561 actttggtgt ctattttctc ttccataaaa aaagcctgac tccacttccc gcgtttactg
4621 attactagcg aagctgcggg tgcatttttt caagataaag gcatccccga ttatattcta
4681 taccgatgtg gattgcgcat actttgtgaa cagaaagtga tagcgttgat gattcttcat
4741 tggtcagaaa attatgaacg gtttcttcta ttttgtctct atatactacg tataggaaat
4801 gtttacattt tcgtattgtt ttcgattcac tctatgaata gttcttacta caattttttt
4861 gtctaaagag taatactaga gataaacata aaaaatgtag aggtcgagtt tagatgcaag
4921 ttcaaggagc gaaaggtgga tgggtaggtt atatagggat atagcacaga gatatatagc
4981 aaagagatac ttttgagcaa tgtttgtgga agcggtattc gcaatatttt agtagctcgt
5041 tacagtccgg tgcgtttttg gttttttgaa agtgcgtctt cagagcgctt ttggttttca
5101 aaagcgctct gaagttccta tactttctag agaataggaa cttcggaata ggaacttcaa
5161 agcgtttccg aaaacgagcg cttccgaaaa tgcaacgcga gctgcgcaca tacagctcac
5221 tgttcacgtc gcacctatat ctgcgtgttg cctgtatata tatatacatg agaagaacgg
5281 catagtgcgt gtttatgctt aaatgcgtac ttatatgcgt ctatttatgt aggatgaaag
5341 gtagtctagt acctcctgtg atattatccc attccatgcg gggtatcgta tgcttccttc
5401 agcactaccc tttagctgtt ctatatgctg ccactcctca attggattag tctcatcctt
5461 caatgctatc atttcctttg atattggatc atattaagaa accattatta tcatgacatt
5521 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
5581 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
5641 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
5701 taactatgcg gcatcagagc agattgtact gagagtgcac catagatcaa cgacattact
5761 atatatataa tataggaagc atttaataga acagcatcgt aatatatgtg tactttgcag
5821 ttatgacgcc agatggcagt agtggaagat attctttatt gaaaaatagc ttgtcacctt
5881 acgtacaatc ttgatccgga gcttttcttt ttttgccgat taagaattaa ttcggtcgaa
5941 aaaagaaaag gagagggcca agagggaggg cattggtgac tattgagcac gtgagtatac
6001 gtgattaagc acacaaaggc agcttggagt atgtctgtta ttaatttcac aggtagttct
6061 ggtccattgg tgaaagtttg cggcttgcag agcacagagg ccgcagaatg tgctctagat
6121 tccgatgctg acttgctggg tattatatgt gtgcccaata gaaagagaac aattgacccg
6181 gttattgcaa ggaaaatttc aagtcttgta aaagcatata aaaatagttc aggcactccg
6241 aaatacttgg ttggcgtgtt tcgtaatcaa cctaaggagg atgttttggc tctggtcaat
6301 gattacggca ttgatatcgt ccaactgcat ggagatgagt cgtggcaaga ataccaagag
6361 ttcctcggtt tgccagttat taaaagactc gtatttccaa aagactgcaa catactactc
6421 agtgcagctt cacagaaacc tcattcgttt attcccttgt ttgattcaga agcaggtggg
6481 acaggtgaac ttttggattg gaactcgatt tctgactggg ttggaaggca agagagcccc
6541 gaaagcttac attttatgtt agctggtgga ctgacgccag aaaatgttgg tgatgcgctt
6601 agattaaatg gcgttattgg tgttgatgta agcggaggtg tggagacaaa tggtgtaaaa
6661 gactctaaca aaatagcaaa tttcgtcaaa aatgctaaga aataggttat tactgagtag
6721 tatttattta agtattgttt gtgcacttgc cgatctatgc ggtgtgaaat accgcacaga
6781 tgcgtaagga gaaaataccg catcaggaaa ttgtaaacgt taatattttg ttaaaattcg
6841 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
6901 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
6961 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
7021 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
7081 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
7141 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
7201 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
7261 cgtccattcg ccattcaggc tgcgcaactg ttgggaaggg cgat
//