LOCUS Exported 5181 bp ds-DNA circular SYN 11-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 5
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5181)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sunday, September 11, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..5181
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 20..36
/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(44..74)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
rep_origin complement(398..986)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1157..2017)
/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(2018..2122)
/gene="bla"
/note="AmpR promoter"
rep_origin 2404..3568
/note="2u ori"
/note="yeast 2u plasmid origin of replication"
promoter 3804..4014
/gene="S. cerevisiae URA3"
/note="URA3 promoter"
CDS 4015..4818
/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"
CDS complement(4891..5181)
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/note="lacZ-alpha"
/translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIG"
primer_bind 5091..5107
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 5111..5167
/note="MCS"
/note="pUC18/19 multiple cloning site"
primer_bind complement(5177..12)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
61 gcataaagtg taaagcctgg ggtgcctaat gagtgaggta actcacatta attgcgttgc
121 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctggattaa tgaatcggcc
181 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
241 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
301 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
361 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
421 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
481 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
541 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac
601 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
661 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
721 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
781 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
841 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
901 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
961 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
1021 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
1081 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
1141 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
1201 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
1261 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
1321 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
1381 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
1441 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
1501 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
1561 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
1621 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
1681 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
1741 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
1801 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
1861 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
1921 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
1981 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
2041 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
2101 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
2161 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
2221 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
2281 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
2341 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
2401 ataacgcatt taagcataaa cacgcactat gccgttcttc tcatgtatat atatatacag
2461 gcaacacgca gatataggtg cgacgtgaac agtgagctgt atgtgcgcag ctcgcgttgc
2521 attttcggaa gcgctcgttt tcggaaacgc tttgaagttc ctattccgaa gttcctattc
2581 tctagctaga aagtatagga acttcagagc gcttttgaaa accaaaagcg ctctgaagac
2641 gcactttcaa aaaaccaaaa acgcaccgga ctgtaacgag ctactaaaat attgcgaata
2701 ccgcttccac aaacattgct caaaagtatc tctttgctat atatctctgt gctatatccc
2761 tatataacct acccatccac ctttcgctcc ttgaacttgc atctaaactc gacctctaca
2821 ttttttatgt ttatctctag tattactctt tagacaaaaa aattgtagta agaactattc
2881 atagagtgaa tcgaaaacaa tacgaaaatg taaacatttc ctatacgtag tatatagaga
2941 caaaatagaa gaaaccgttc ataattttct gaccaatgaa gaatcatcaa cgctatcact
3001 ttctgttcac aaagtatgcg caatccacat cggtatagaa tataatcggg gatgccttta
3061 tcttgaaaaa atgcacccgc agcttcgcta gtaatcagta aacgcgggaa gtggagtcag
3121 gcttttttta tggaagagaa aatagacacc aaagtagcct tcttctaacc ttaacggacc
3181 tacagtgcaa aaagttatca agagactgca ttatagagcg cacaaaggag aaaaaaagta
3241 atctaagatg ctttgttaga aaaatagcgc tctcgggatg catttttgta gaacaaaaaa
3301 gaagtataga ttctttgttg gtaaaatagc gctctcgcgt tgcatttctg ttctgtaaaa
3361 atgcagctca gattctttgt ttgaaaaatt agcgctctcg cgttgcattt ttgttttaca
3421 aaaatgaagc acagattctt cgttggtaaa atagcgcttt cgcgttgcat ttctgttctg
3481 taaaaatgca gctcagattc tttgtttgaa aaattagcgc tctcgcgttg catttttgtt
3541 ctacaaaatg aagcacagat gcttcgttaa caaagatatg ctattgaagt gcaagatgga
3601 aacgcagaaa atgaaccggg gatgcgacgt gcaagattac ctatgcaata gatgcaatag
3661 tttctccagg aaccgaaata catacattgt cttccgtaaa gcgctagact atatattatt
3721 atacaggttc aaatatacta tctgtttcag ggaaaactcc caggttcgga tgttcaaaat
3781 tcaatgatgg gtaacaagag cttttcaatt catcattttt tttttattct tttttttgat
3841 ttcggtttct ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga
3901 aggagcacag acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt
3961 gcccagtatt cttaacccaa ctgcacagaa caaaaaccga aacgaagata aatcatgtcg
4021 aaagctacat ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt
4081 aatatcatgc acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag
4141 gaattactgg agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg
4201 gatatcttga ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc
4261 aagtacaatt ttttactctt cgaagacaga aaatttgctg acattggtaa tacagtcaaa
4321 ttgcagtact ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac
4381 ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag
4441 gaacctagag gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga
4501 gaatatacta agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt
4561 attgctcaaa gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc
4621 ggtgtgggtt tagatgacaa gggagacgca ttgggtcaac agtatagaac cgtggatgat
4681 gtggtctcta caggatctga cattattatt gttggaagag gactatttgc aaagggaagg
4741 gatgctaagg tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga
4801 tgcggccagc aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa
4861 attagagctt caatttaatt atatcagtta ttaccctatg cggtgtgaaa taccgcacag
4921 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
4981 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaagg ggggatgtgc
5041 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
5101 ggccagtgcc aagcttgcat gcctgcaggt cgactctaga ggatccccgg gtaccgagct
5161 cgaattcgta atcatggtca t
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