LOCUS Exported 5240 bp ds-DNA circular SYN 06-JAN-2018
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..5240
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 106..127
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 142..172
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 180..196
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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 204..220
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
CDS 216..536
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIIGSYLLPII
EF"
misc_feature 233..289
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(290..306)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 742..2088
/label=2u ori
/note="yeast 2u plasmid origin of replication"
CDS complement(2199..3002)
/codon_start=1
/gene="S. cerevisiae URA3"
/product="orotidine-5'-phosphate decarboxylase, required
for uracil biosynthesis"
/label=URA3
/note="yeast auxotrophic marker, counterselectable with
5-fluoroorotic acid (5-FOA)"
/translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
promoter complement(3003..3218)
/gene="S. cerevisiae URA3"
/label=URA3 promoter
promoter 3334..3438
/gene="bla"
/label=AmpR promoter
CDS 3439..4299
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 4470..5058
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
241 atgcctgcag gtcgactcta gaggatcccc gggtaccgag ctcgaattca ctggccgtcg
301 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
361 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
421 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt
481 gcggtatttc acaccgcata atcggatcgt acttgttacc catcattgaa ttttgaacat
541 ccgaacctgg gagttttccc tgaaacagat agtatatttg aacctgtata ataatatata
601 gtctagcgct ttacggaaga caatgtatgt atttcggttc ctggagaaac tattgcatct
661 attgcatagg taatcttgca cgtcgcatcc ccggttcatt ttctgcgttt ccatcttgca
721 cttcaatagc atatctttgt taacgaagca tctgtgcttc attttgtaga acaaaaatgc
781 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
841 tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt gtaaaacaaa
901 aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac
961 agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct
1021 acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc
1081 tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag
1141 gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact
1201 tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc
1261 ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt
1321 tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac
1381 tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt
1441 actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg
1501 agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca
1561 cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaata
1621 ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg tcttcagagc
1681 gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag aataggaact
1741 tcggaatagg aacttcaaag cgtttccgaa aacgagcgct tccgaaaatg caacgcgagc
1801 tgcgcacata cagctcactg ttcacgtcgc acctatatct gcgtgttgcc tgtatatata
1861 tatacatgag aagaacggca tagtgcgtgt ttatgcttaa atgcgtactt atatgcgtct
1921 atttatgtag gatgaaaggt agtctagtac ctcctgtgat attatcccat tccatgcggg
1981 gtatcgtatg cttccttcag cactaccctt tagctgttct atatgctgcc actcctcaat
2041 tggattagtc tcatccttca atgctatcat ttcctttgat attggatcga tccgatgata
2101 agctgtcaaa catgagaatt gggtaataac tgatataatt aaattgaagc tctaatttgt
2161 gagtttagta tacatgcatt tacttataat acagtttttt agttttgctg gccgcatctt
2221 ctcaaatatg cttcccagcc tgcttttctg taacgttcac cctctacctt agcatccctt
2281 ccctttgcaa atagtcctct tccaacaata ataatgtcag atcctgtaga gaccacatca
2341 tccacggttc tatactgttg acccaatgcg tctcccttgt catctaaacc cacaccgggt
2401 gtcataatca accaatcgta accttcatct cttccaccca tgtctctttg agcaataaag
2461 ccgataacaa aatctttgtc gctcttcgca atgtcaacag tacccttagt atattctcca
2521 gtagataggg agcccttgca tgacaattct gctaacatca aaaggcctct aggttccttt
2581 gttacttctt ctgccgcctg cttcaaaccg ctaacaatac ctgggcccac cacaccgtgt
2641 gcattcgtaa tgtctgccca ttctgctatt ctgtatacac ccgcagagta ctgcaatttg
2701 actgtattac caatgtcagc aaattttctg tcttcgaaga gtaaaaaatt gtacttggcg
2761 gataatgcct ttagcggctt aactgtgccc tccatggaaa aatcagtcaa gatatccaca
2821 tgtgttttta gtaaacaaat tttgggacct aatgcttcaa ctaactccag taattccttg
2881 gtggtacgaa catccaatga agcacacaag tttgtttgct tttcgtgcat gatattaaat
2941 agcttggcag caacaggact aggatgagta gcagcacgtt ccttatatgt agctttcgac
3001 atgatttatc ttcgtttcct gcatgttttt gttctgtgca gttgggttaa gaatactggg
3061 caatttcatg tttcttcaac actacatatg cgtatatata ccaatctaag tctgtgctcc
3121 ttccttcgtt cttccttctg ttcggagatt accgaatcaa aaaaatttca aagaaaccga
3181 aatcaaaaaa aagaataaaa aaaaaatgat gaattgaatt gaaaagctaa ttcttgaaga
3241 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
3301 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
3361 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
3421 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
3481 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
3541 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
3601 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
3661 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
3721 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
3781 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
3841 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
3901 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
3961 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
4021 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
4081 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
4141 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
4201 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
4261 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
4321 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
4381 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
4441 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
4501 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
4561 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
4621 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
4681 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
4741 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
4801 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
4861 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
4921 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
4981 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
5041 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
5101 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
5161 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
5221 gtgagcgagg aagcggaaga
//