LOCUS Exported 6849 bp ds-DNA circular SYN 12-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 7
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6849)
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..6849
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(663..1757)
/codon_start=1
/gene="S. cerevisiae LEU2"
/product="3-isopropylmalate dehydrogenase, required for
leucine biosynthesis"
/note="LEU2"
/note="yeast auxotrophic marker"
/translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
IGGAAIDATGVPLPDEALEASKKVDAVLLGAVAGPKWGTGSVRPEQGLLKIRKELQLYA
NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
VPEVQRITRMAAFMALQHEPPLPIWSLDKANLLASSRLWRKTVEETIKNEFPTLKVQHQ
LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
DLGGSNSTTEVGDAVAEEVKKILA"
promoter complement(1770..2174)
/gene="S. cerevisiae LEU2"
/note="LEU2 promoter"
rep_origin complement(2474..2929)
/direction=LEFT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(2712..3290)
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/note="lacZ-alpha"
/translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
TDRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFA
GFPRQALNRGLPLGFRFSALRHLDPKKLD"
primer_bind 3074..3090
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter 3100..3118
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 3127..3234
/note="MCS"
/note="pBluescript multiple cloning site"
primer_bind 3144..3160
/note="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3194..3210)
/note="SK primer"
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(3247..3265)
/note="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(3286..3302)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 3310..3326
/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(3334..3364)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
rep_origin complement(3688..4276)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4447..5307)
/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(5308..5412)
/gene="bla"
/note="AmpR promoter"
rep_origin 5439..6781
/note="2u ori"
/note="yeast 2u plasmid origin of replication"
protein_bind complement(6402..6449)
/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 accatatcga ctacgtcgta aggccgtttc tgacagagta aaattcttga gggaactttc
241 accattatgg gaaatgcttc aagaaggtat tgacttaaac tccatcaaat ggtcaggtca
301 ttgagtgttt tttatttgtt gtattttttt ttttttagag aaaatcctcc aatatcaaat
361 taggaatcgt agtttcatga ttttctgtta cacctaactt tttgtgtggt gccctcctcc
421 ttgtcaatat taatgttaaa gtgcaattct ttttccttat cacgttgagc cattagtatc
481 aatttgctta cctgtattcc tttactatcc tcctttttct ccttcttgat aaatgtatgt
541 agattgcgta tatagtttcg tctaccctat gaacatattc cattttgtaa tttcgtgtcg
601 tttctattat gaatttcatt tataaagttt atgtacaaat atcataaaaa aagagaatct
661 ttttaagcaa ggattttctt aacttcttcg gcgacagcat caccgacttc ggtggtactg
721 ttggaaccac ctaaatcacc agttctgata cctgcatcca aaaccttttt aactgcatct
781 tcaatggcct taccttcttc aggcaagttc aatgacaatt tcaacatcat tgcagcagac
841 aagatagtgg cgatagggtc aaccttattc tttggcaaat ctggagcaga accgtggcat
901 ggttcgtaca aaccaaatgc ggtgttcttg tctggcaaag aggccaagga cgcagatggc
961 aacaaaccca aggaacctgg gataacggag gcttcatcgg agatgatatc accaaacatg
1021 ttgctggtga ttataatacc atttaggtgg gttgggttct taactaggat catggcggca
1081 gaatcaatca attgatgttg aaccttcaat gtagggaatt cgttcttgat ggtttcctcc
1141 acagtttttc tccataatct tgaagaggcc aaaagattag ctttatccaa ggaccaaata
1201 ggcaatggtg gctcatgttg tagggccatg aaagcggcca ttcttgtgat tctttgcact
1261 tctggaacgg tgtattgttc actatcccaa gcgacaccat caccatcgtc ttcctttctc
1321 ttaccaaagt aaatacctcc cactaattct ctgacaacaa cgaagtcagt acctttagca
1381 aattgtggct tgattggaga taagtctaaa agagagtcgg atgcaaagtt acatggtctt
1441 aagttggcgt acaattgaag ttctttacgg atttttagta aaccttgttc aggtctaaca
1501 ctaccggtac cccatttagg accagccaca gcacctaaca aaacggcatc aaccttcttg
1561 gaggcttcca gcgcctcatc tggaagtggg acacctgtag catcgatagc agcaccacca
1621 attaaatgat tttcgaaatc gaacttgaca ttggaacgaa catcagaaat agctttaaga
1681 accttaatgg cttcggctgt gatttcttga ccaacgtggt cacctggcaa aacgacgatc
1741 ttcttagggg cagacatagg ggcagacatt agaatggtat atccttgaaa tatatatata
1801 tattgctgaa atgtaaaagg taagaaaagt tagaaagtaa gacgattgct aaccacctat
1861 tggaaaaaac aataggtcct taaataatat tgtcaacttc aagtattgtg atgcaagcat
1921 ttagtcatga acgcttctct attctatatg aaaagccggt tccggcctct cacctttcct
1981 ttttctccca atttttcagt tgaaaaaggt atatgcgtca ggcgacctct gaaattaaca
2041 aaaaatttcc agtcatcgaa tttgattctg tgcgatagcg cccctgtgtg ttctcgttat
2101 gttgaggaaa aaaataatgg ttgctaagag attcgaactc ttgcatctta cgatacctga
2161 gtattcccac agttaactgc ggtcaagata tttcttgaat caggcgcctt agaccgctcg
2221 gccaaacaac caattacttg ttgagaaata gagtataatt atcctataaa tataacgttt
2281 ttgaacacac atgaacaagg aagtacagga caattgattt tgaagagaat gtggattttg
2341 atgtaattgt tgggattcca tttttaataa ggcaataata ttaggtatgt ggatatacta
2401 gaagttctcc tcgaccgtcg atatgcggtg tgaaataccg cacagatgcg taaggagaaa
2461 ataccgcatc aggaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt
2521 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
2581 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
2641 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
2701 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
2761 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
2821 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
2881 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc gcgccattcg
2941 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
3001 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
3061 cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc
3121 gaattgggta ccgggccccc cctcgaggtc gacggtatcg ataagcttga tatcgaattc
3181 ctgcagcccg ggggatccac tagttctaga gcggccgcca ccgcggtgga gctccagctt
3241 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
3301 tgtgtgaaat tgttatccgc tcacaattcc acacaacata ggagccggaa gcataaagtg
3361 taaagcctgg ggtgcctaat gagtgaggta actcacatta attgcgttgc gctcactgcc
3421 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
3481 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
3541 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
3601 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
3661 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
3721 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
3781 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
3841 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
3901 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
3961 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
4021 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
4081 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
4141 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
4201 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
4261 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
4321 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
4381 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
4441 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
4501 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
4561 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
4621 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
4681 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
4741 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
4801 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
4861 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
4921 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
4981 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
5041 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
5101 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
5161 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
5221 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
5281 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
5341 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
5401 aggggttccg cgcacatttc cccgaaaagt gccacctgaa cgaagcatct gtgcttcatt
5461 ttgtagaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca
5521 tttttacaga acagaaatgc aacgcgaaag cgctatttta ccaacgaaga atctgtgctt
5581 catttttgta aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag
5641 ctgcattttt acagaacaga aatgcaacgc gagagcgcta ttttaccaac aaagaatcta
5701 tacttctttt ttgttctaca aaaatgcatc ccgagagcgc tatttttcta acaaagcatc
5761 ttagattact ttttttctcc tttgtgcgct ctataatgca gtctcttgat aactttttgc
5821 actgtaggtc cgttaaggtt agaagaaggc tactttggtg tctattttct cttccataaa
5881 aaaagcctga ctccacttcc cgcgtttact gattactagc gaagctgcgg gtgcattttt
5941 tcaagataaa ggcatccccg attatattct ataccgatgt ggattgcgca tactttgtga
6001 acagaaagtg atagcgttga tgattcttca ttggtcagaa aattatgaac ggtttcttct
6061 attttgtctc tatatactac gtataggaaa tgtttacatt ttcgtattgt tttcgattca
6121 ctctatgaat agttcttact acaatttttt tgtctaaaga gtaatactag agataaacat
6181 aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt
6241 tatataggga tatagcacag agatatatag caaagagata cttttgagca atgtttgtgg
6301 aagcggtatt cgcaatattt tagtagctcg ttacagtccg gtgcgttttt ggttttttga
6361 aagtgcgtct tcagagcgct tttggttttc aaaagcgctc tgaagttcct atactttcta
6421 gagaatagga acttcggaat aggaacttca aagcgtttcc gaaaacgagc gcttccgaaa
6481 atgcaacgcg agctgcgcac atacagctca ctgttcacgt cgcacctata tctgcgtgtt
6541 gcctgtatat atatatacat gagaagaacg gcatagtgcg tgtttatgct taaatgcgta
6601 cttatatgcg tctatttatg taggatgaaa ggtagtctag tacctcctgt gatattatcc
6661 cattccatgc ggggtatcgt atgcttcctt cagcactacc ctttagctgt tctatatgct
6721 gccactcctc aattggatta gtctcatcct tcaatgctat catttccttt gatattggat
6781 catactaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
6841 cctttcgtc
//