LOCUS Exported 7693 bp ds-DNA circular SYN 29-DEC-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..7693
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 69..1411
/label=2u ori
/note="yeast 2u plasmid origin of replication"
protein_bind 401..448
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
promoter 1438..1542
/gene="bla"
/label=AmpR promoter
CDS 1543..2403
/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 2574..3162
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 3626..4032
/gene="RPR1"
/label=RPR1 promoter
/note="promoter for the S. cerevisiae RNase P RNA gene"
misc_RNA 4040..4115
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
terminator 4128..4190
/label=RPR1 terminator
/note="transcription terminator for the S. cerevisiae RNase
P RNA gene"
rep_origin 4765..5220
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 5520..5924
/gene="S. cerevisiae LEU2"
/label=LEU2 promoter
CDS 5937..7031
/codon_start=1
/gene="S. cerevisiae LEU2"
/product="3-isopropylmalate dehydrogenase, required for
leucine biosynthesis"
/label=LEU2
/note="yeast auxotrophic marker"
/translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
IGGAAIDATGVPLPDEALEASKKVDAVLLGAVAGPKWGTGSVRPEQGLLKIRKELQLYA
NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
VPEVQRITRMAAFMALQHEPPLPIWSLDKANLLASSRLWRKTVEETIKNEFPTLKVQHQ
LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
DLGGSNSTTEVGDAVAEEVKKILA"
ORIGIN
1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
61 cttagtatga tccaatatca aaggaaatga tagcattgaa ggatgagact aatccaattg
121 aggagtggca gcatatagaa cagctaaagg gtagtgctga aggaagcata cgataccccg
181 catggaatgg gataatatca caggaggtac tagactacct ttcatcctac ataaatagac
241 gcatataagt acgcatttaa gcataaacac gcactatgcc gttcttctca tgtatatata
301 tatacaggca acacgcagat ataggtgcga cgtgaacagt gagctgtatg tgcgcagctc
361 gcgttgcatt ttcggaagcg ctcgttttcg gaaacgcttt gaagttccta ttccgaagtt
421 cctattctct agaaagtata ggaacttcag agcgcttttg aaaaccaaaa gcgctctgaa
481 gacgcacttt caaaaaacca aaaacgcacc ggactgtaac gagctactaa aatattgcga
541 ataccgcttc cacaaacatt gctcaaaagt atctctttgc tatatatctc tgtgctatat
601 ccctatataa cctacccatc cacctttcgc tccttgaact tgcatctaaa ctcgacctct
661 acatttttta tgtttatctc tagtattact ctttagacaa aaaaattgta gtaagaacta
721 ttcatagagt gaatcgaaaa caatacgaaa atgtaaacat ttcctatacg tagtatatag
781 agacaaaata gaagaaaccg ttcataattt tctgaccaat gaagaatcat caacgctatc
841 actttctgtt cacaaagtat gcgcaatcca catcggtata gaatataatc ggggatgcct
901 ttatcttgaa aaaatgcacc cgcagcttcg ctagtaatca gtaaacgcgg gaagtggagt
961 caggcttttt ttatggaaga gaaaatagac accaaagtag ccttcttcta accttaacgg
1021 acctacagtg caaaaagtta tcaagagact gcattataga gcgcacaaag gagaaaaaaa
1081 gtaatctaag atgctttgtt agaaaaatag cgctctcggg atgcattttt gtagaacaaa
1141 aaagaagtat agattctttg ttggtaaaat agcgctctcg cgttgcattt ctgttctgta
1201 aaaatgcagc tcagattctt tgtttgaaaa attagcgctc tcgcgttgca tttttgtttt
1261 acaaaaatga agcacagatt cttcgttggt aaaatagcgc tttcgcgttg catttctgtt
1321 ctgtaaaaat gcagctcaga ttctttgttt gaaaaattag cgctctcgcg ttgcattttt
1381 gttctacaaa atgaagcaca gatgcttcgt tcaggtggca cttttcgggg aaatgtgcgc
1441 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
1501 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
1561 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
1621 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
1681 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
1741 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
1801 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
1861 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
1921 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
1981 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
2041 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
2101 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
2161 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
2221 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
2281 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
2341 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
2401 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
2461 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
2521 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
2581 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
2641 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
2701 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
2761 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
2821 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
2881 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
2941 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
3001 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
3061 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
3121 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
3181 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
3241 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
3301 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
3361 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
3421 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt acctcactca ttaggcaccc
3481 caggctttac actttatgct tccggctcct atgttgtgtg gaattgtgag cggataacaa
3541 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
3601 agggaacaaa agctggagct cgggggatct gccaattgaa cataacatgg tagttacata
3661 tactagtaat atggttcggc acacattaaa agtataaaaa ctatctgaat tacgaattac
3721 atatattggt cataaaaatc aatcaatcat cgtgtgtttt atatgtctct tatctaagta
3781 taagaatatc catagttaat attcacttac gctacctttt aacctgtaat cattgtcaac
3841 aggatatgtt aacgacccac attgataaac gctagtattt ctttttcctc ttcttattgg
3901 ccggctgtct ctatactccc ctatagtctg tttcttttcg tttcgattgt tttacgtttg
3961 aggcctcgtg gcgcacatgg tacgctgtgg tgctcgcggc tgggaacgaa actctgggag
4021 ctgcgattgg cagaagcttg ttttagagct agaaatagca agttaaaata aggctagtcc
4081 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tctcgagcca tatccaactt
4141 ccaatttaat ctttcttttt taattttcac ttatttgcga tacagaaaga aaaaagcgat
4201 agtaactatt gaattttgtt tggatttggt tagattagat atggtttctc tttatattta
4261 catgctaaaa atgggctaca ccagagatac ataattagat atatatacgc cagtacacct
4321 tatcggccca agccttgtcc caaggcagcg ttttgttctt ggaaacgctg ccctacacgt
4381 tcgctatgct tcaagaactt ttctgagcac ttcatgatgc atgtttgttc cttattggtt
4441 agctttgatg ttgtgaagtc attgacacag tctgtgaaac atctttctac cagattagag
4501 tacaaacgca tgaaatcctt catttgcttt tgttccacta ctttttggaa ctcttgttgt
4561 tctttggtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt
4621 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
4681 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
4741 ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
4801 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
4861 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
4921 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
4981 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
5041 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
5101 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
5161 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
5221 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatcgacggt
5281 cgaggagaac ttctagtata tccacatacc taatattatt gccttattaa aaatggaatc
5341 ccaacaatta catcaaaatc cacattctct tcaaaatcaa ttgtcctgta cttccttgtt
5401 catgtgtgtt caaaaacgtt atatttatag gataattata ctctatttct caacaagtaa
5461 ttggttgttt ggccgagcgg tctaaggcgc ctgattcaag aaatatcttg accgcagtta
5521 actgtgggaa tactcaggta tcgtaagatg caagagttcg aatctcttag caaccattat
5581 ttttttcctc aacataacga gaacacacag gggcgctatc gcacagaatc aaattcgatg
5641 actggaaatt ttttgttaat ttcagaggtc gcctgacgca tatacctttt tcaactgaaa
5701 aattgggaga aaaaggaaag gtgagaggcc ggaaccggct tttcatatag aatagagaag
5761 cgttcatgac taaatgcttg catcacaata cttgaagttg acaatattat ttaaggacct
5821 attgtttttt ccaataggtg gttagcaatc gtcttacttt ctaacttttc ttacctttta
5881 catttcagca atatatatat atatttcaag gatataccat tctaatgtct gcccctatgt
5941 ctgcccctaa gaagatcgtc gttttgccag gtgaccacgt tggtcaagaa atcacagccg
6001 aagccattaa ggttcttaaa gctatttctg atgttcgttc caatgtcaag ttcgatttcg
6061 aaaatcattt aattggtggt gctgctatcg atgctacagg tgtcccactt ccagatgagg
6121 cgctggaagc ctccaagaag gttgatgccg ttttgttagg tgctgtggct ggtcctaaat
6181 ggggtaccgg tagtgttaga cctgaacaag gtttactaaa aatccgtaaa gaacttcaat
6241 tgtacgccaa cttaagacca tgtaactttg catccgactc tcttttagac ttatctccaa
6301 tcaagccaca atttgctaaa ggtactgact tcgttgttgt cagagaatta gtgggaggta
6361 tttactttgg taagagaaag gaagacgatg gtgatggtgt cgcttgggat agtgaacaat
6421 acaccgttcc agaagtgcaa agaatcacaa gaatggccgc tttcatggcc ctacaacatg
6481 agccaccatt gcctatttgg tccttggata aagctaatct tttggcctct tcaagattat
6541 ggagaaaaac tgtggaggaa accatcaaga acgaattccc tacattgaag gttcaacatc
6601 aattgattga ttctgccgcc atgatcctag ttaagaaccc aacccaccta aatggtatta
6661 taatcaccag caacatgttt ggtgatatca tctccgatga agcctccgtt atcccaggtt
6721 ccttgggttt gttgccatct gcgtccttgg cctctttgcc agacaagaac accgcatttg
6781 gtttgtacga accatgccac ggttctgctc cagatttgcc aaagaataag gttgacccta
6841 tcgccactat cttgtctgct gcaatgatgt tgaaattgtc attgaacttg cctgaagaag
6901 gtaaggccat tgaagatgca gttaaaaagg ttttggatgc aggtatcaga actggtgatt
6961 taggtggttc caacagtacc accgaagtcg gtgatgctgt cgccgaagaa gttaagaaaa
7021 tccttgctta aaaagattct ctttttttat gatatttgta cataaacttt ataaatgaaa
7081 ttcataatag aaacgacacg aaattacaaa atggaatatg ttcatagggt agacgaaact
7141 atatacgcaa tctacataca tttatcaaga aggagaaaaa ggaggatagt aaaggaatac
7201 aggtaagcaa attgatacta atggctcaac gtgataagga aaaagaattg cactttaaca
7261 ttaatattga caaggaggag ggcaccacac aaaaagttag gtgtaacaga aaatcatgaa
7321 actacgattc ctaatttgat attggaggat tttctctaaa aaaaaaaaaa tacaacaaat
7381 aaaaaacact caatgacctg accatttgat ggagtttaag tcaatacctt cttgaagcat
7441 ttcccataat ggtgaaagtt ccctcaagaa ttttactctg tcagaaacgg ccttacgacg
7501 tagtcgatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
7561 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
7621 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
7681 ccgaaacgcg cga
//