LOCUS Exported 5025 bp ds-DNA circular SYN 19-JAN-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..5025
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 84..100
/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)."
CDS 271..288
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
misc_feature 289..840
/label=ProS2
CDS 850..873
/codon_start=1
/product="recognition and cleavage site for human
rhinovirus 3C and PreScission proteases"
/label=HRV 3C site
/translation="LEVLFQGP"
CDS 883..900
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
CDS 907..918
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
rep_origin complement(1329..1784)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 1884..1988
/gene="bla"
/label=AmpR promoter
CDS 1989..2849
/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 3020..3608
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3780..4862)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
promoter complement(4863..4940)
/gene="lacI"
/label=lacI promoter
ORIGIN
1 aaggaatggt gtggccgatt aatcataaat atgaaaaata attgttgcat cacccgccaa
61 tgcgtggctt aatgcacatc aaattgtgag cggataacaa tttgatgtgc tagcgcatat
121 ccagtgtagt aaggcaagtc ccttcaagag ttatcgttga tacccctcgt agtgcacatt
181 cctttaacgc ttcaaaatct gtaaagcacg ccatatcgcc gaaaggcaca cttaattatt
241 aagaggtaat acaccatgaa tcacaaagtg catcatcatc atcatcacat ggcaaatatt
301 accgttttct ataacgaaga cttccagggt aagcaggtcg atctgccgcc tggcaactat
361 acccgcgccc agttggcggc gctgggcatc gagaataata ccatcagctc ggtgaaggtg
421 ccgcctggcg tgaaggctat cctgtaccag aacgatggtt tcgccggcga ccagatcgaa
481 gtggtggcca atgccgagga gttgggcccg ctgaataata acgtctccag catccgcgtc
541 atctccgtgc ccgtgcagcc gcgcatggca aatattaccg ttttctataa cgaagacttc
601 cagggtaagc aggtcgatct gccgcctggc aactataccc gcgcccagtt ggcggcgctg
661 ggcatcgaga ataataccat cagctcggtg aaggtgccgc ctggcgtgaa ggctatcctc
721 taccagaacg atggtttcgc cggcgaccag atcgaagtgg tggccaatgc cgaggagctg
781 ggtccgctga ataataacgt ctccagcatc cgcgtcatct ccgtgccggt gcagccgagg
841 tccgcgggtc tggaagttct gttccagggg ccctccgcgg gtctggtgcc acgcggtagt
901 ggtggtatcg aaggtaggca tatggagctc ggtaccctcg agggatccga attcaagctt
961 gtcgacctgc agtctagata ggtaatctct gcttaaaagc acagaatcta agatccctgc
1021 catttggcgg ggattttttt atttgttttc aggaaataaa taatcgatcg cgtaataaaa
1081 tctattatta tttttgtgaa gaataaattt gggtgcaatg agaatgcgca ggccctttcg
1141 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
1201 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
1261 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
1321 gcaccataaa attgtaaacg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
1381 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
1441 gcccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
1501 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
1561 atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
1621 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
1681 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
1741 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtactatg gttgctttga
1801 cgtatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgtcag
1861 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
1921 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
1981 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
2041 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
2101 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
2161 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
2221 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
2281 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
2341 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
2401 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
2461 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
2521 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
2581 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
2641 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
2701 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
2761 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
2821 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
2881 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
2941 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
3001 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
3061 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
3121 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
3181 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
3241 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
3301 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
3361 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
3421 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
3481 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
3541 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
3601 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
3661 ctcacatagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
3721 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
3781 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
3841 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
3901 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
3961 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
4021 catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc
4081 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
4141 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
4201 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
4261 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
4321 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
4381 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
4441 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
4501 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
4561 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
4621 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
4681 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
4741 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
4801 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
4861 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
4921 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag
4981 gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgc
//