LOCUS Exported 4887 bp ds-DNA circular SYN 12-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4887)
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..4887
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 70..573
/note="CEN/ARS"
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
promoter 610..714
/gene="bla"
/note="AmpR promoter"
CDS 715..1575
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 1746..2334
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 2658..2688
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 2696..2712
/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 2720..2736
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
CDS 2732..3307
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/note="lacZ-alpha"
/translation="MTMITPSSELTLTKGNKSWVPGPPSRSTVSISLISNSCSPGDPLV
LERPPPRWSSNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
DRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFAG
FPRQALNRGLPLGFRFSALRHLDPKKLD"
promoter 2757..2775
/note="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
misc_feature 2788..2895
/note="MCS"
/note="pBluescript multiple cloning site"
primer_bind 2805..2821
/note="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2855..2871)
/note="SK primer"
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(2904..2922)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2929..2945)
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 3090..3545
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(3674..4477)
/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"
promoter complement(4478..4693)
/gene="S. cerevisiae URA3"
/note="URA3 promoter"
ORIGIN
1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
61 cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata
121 atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt
181 aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa
241 atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata
301 gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct
361 tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata
421 aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact
481 attttttctt taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa
541 atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg cacttttcgg
601 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
661 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
721 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
781 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
841 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
901 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
961 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
1021 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
1081 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
1141 ccgaaggagc taaccgcttt ttttcacaac atgggggatc atgtaactcg ccttgatcgt
1201 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
1261 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
1321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
1381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
1441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
1501 ggcagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
1561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
1621 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
1681 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
1741 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
1801 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
1861 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
1921 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
1981 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
2041 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
2101 acgacctaca ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc
2161 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
2221 agggagcttc caggggggaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
2281 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc cgagcctatg gaaaaacgcc
2341 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
2401 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
2461 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
2521 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
2581 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact
2641 cattaggcac cccaggcttt acactttatg cttccggctc ctatgttgtg tggaattgtg
2701 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctcggaatt
2761 aaccctcact aaagggaaca aaagctgggt accgggcccc ccctcgaggt cgacggtatc
2821 gataagcttg atatcgaatt cctgcagccc gggggatcca ctagttctag agcggccgcc
2881 accgcggtgg agctccaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt
2941 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
3001 tccccccttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
3061 gttgcgcagc ctgaatggcg aatggcgcga cgcgccctgt agcggcgcat taagcgcggc
3121 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
3181 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
3241 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
3301 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
3361 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
3421 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
3481 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
3541 aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcagggta
3601 ataactgata taattaaatt gaagctctaa tttgtgagtt tagtatacat gcatttactt
3661 ataatacagt tttttagttt tgctggccgc atcttctcaa atatgcttcc cagcctgctt
3721 ttctgtaacg ttcaccctct accttagcat cccttccctt tgcaaatagt cctcttccaa
3781 caataataat gtcagatcct gtagagacca catcatccac ggttctatac tgttgaccca
3841 atgcgtctcc cttgtcatct aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt
3901 catctcttcc acccatgtct ctttgagcaa taaagccgat aacaaaatct ttgtcgctct
3961 tcgcaatgtc aacagtaccc ttagtatatt ctccagtaga tagggagccc ttgcatgaca
4021 attctgctaa catcaaaagg cctctaggtt cctttgttac ttcttctgcc gcctgcttca
4081 aaccgctaac aatacctggg cccaccacac cgtgtgcatt cgtaatgtct gcccattctg
4141 ctattctgta tacacccgca gagtactgca atttgactgt attaccaatg tcagcaaatt
4201 ttctgtcttc gaagagtaaa aaattgtact tggcggataa tgcctttagc ggcttaactg
4261 tgccctccat ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa caaattttgg
4321 gacctaatgc ttcaactaac tccagtaatt ccttggtggt acgaacatcc aatgaagcac
4381 acaagtttgt ttgcttttcg tgcatgatat taaatagctt ggcagcaaca ggactaggat
4441 gagtagcagc acgttcctta tatgtagctt tcgacatgat ttatcttcgt ttcctgcagg
4501 tttttgttct gtgcagttgg gttaagaata ctgggcaatt tcatgtttct tcaacactac
4561 atatgcgtat atataccaat ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg
4621 agattaccga atcaaaaaaa tttcaaagaa accgaaatca aaaaaaagaa taaaaaaaaa
4681 atgatgaatt gaattgaaaa gcgtggtgca ctctcagtac aatctgctct gatgccgcat
4741 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
4801 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
4861 tttcaccgtc atcaccgaaa cgcgcga
//