LOCUS Exported 5097 bp ds-DNA circular SYN 06-JUL-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..5097
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 86..163
/gene="lacI"
/label=lacI promoter
CDS 164..1246
/codon_start=1
/gene="lacI"
/product="lac repressor"
/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"
rep_origin complement(1418..2006)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2177..3037)
/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"
promoter complement(3038..3142)
/gene="bla"
/label=AmpR promoter
rep_origin 3242..3697
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(4111..4134)
/codon_start=1
/product="recognition and cleavage site for human
rhinovirus 3C and PreScission proteases"
/label=HRV 3C site
/translation="LEVLFQGP"
CDS complement(4141..4794)
/codon_start=1
/product="glutathione S-transferase from Schistosoma
japonicum"
/label=GST
/translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
CDS complement(4798..4809)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
CDS complement(4810..4827)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
protein_bind 4998..5014
/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)."
ORIGIN
1 gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg
61 cataagggag agcgtcgaga tcccggacac catcgaatgg cgcaaaacct ttcgcggtat
121 ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt
181 atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca
241 ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa
301 ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt
361 tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg
421 cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc
481 ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta
541 tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt
601 atttcttgat gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg
661 tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc
721 gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac
781 tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt
841 tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa
901 cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc
961 ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt
1021 aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca
1081 actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag
1141 aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
1201 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
1261 atgtaagtta gctcactcat taggcaccgg gatctcgacc gatgcccttg agagccttca
1321 acccagtcag ctccttccgg tgggcgcggg gcatgactat gtgagcaaaa ggccagcaaa
1381 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
1441 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
1501 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
1561 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
1621 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
1681 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
1741 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
1801 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
1861 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
1921 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
1981 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
2041 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
2101 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
2161 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
2221 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
2281 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
2341 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
2401 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
2461 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
2521 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
2581 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
2641 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
2701 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
2761 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
2821 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
2881 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
2941 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
3001 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
3061 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
3121 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gcctgatgcg
3181 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
3241 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
3301 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
3361 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
3421 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
3481 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
3541 ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta
3601 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
3661 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
3721 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
3781 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
3841 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctgcgc
3901 attctcattg cacccaaatt tattcttcac aaaaataata atagatttta ttacgcgatc
3961 gattatttat ttcctgaaaa caaataaaaa aatccccgcc aaatggcagg gatcttagat
4021 tctgtgcttt taagcagaga ttacctatct agactgcagg tcgacaagct tgaattcgga
4081 tccctcgagg gtaccgagct ccatatgtcc cgggccctgg aacagaactt ccagatccga
4141 ttttggagga tggtcgccac caccaaacgt ggcttgccag ccctgcaaag gccatgctat
4201 atacttgctg gatttcaagt acttatcaat ttgtgggata gcttcaatac gttttttaaa
4261 acaaactaat tttgggaacg catccaggca cattgggtcc atgtataaaa caacatcaag
4321 agcgtcatac aacatgaagt caggatgggt tacatgatca ccatttaaat atgttttatg
4381 acataaacga tcttcgaaca ttttcagcat ttcaggtagc ttgctaagaa aatcaacttt
4441 gagagtttca aagtctttac tatatgcaat tctcgaaaca ccgtatctaa tatccaaaac
4501 cgctccttca agcattgaaa tctctgcacg ctcttttgga caaccaccca acatgttgtg
4561 cttgtcagct atataacgta tgatggccat agactgtgtt aatttaacat caccatcaat
4621 ataataagga agattgggaa actccaaacc caattcaaac tttttgtttc gccatttatc
4681 accttcatcg cgctcataca aatgctcttc atatttttct tcaagatatt ccaaaagaag
4741 tcgagtgggt tgcacaaggc ccttaatttt ccaataacct agtatagggg acatgtgcct
4801 accttcgata tgatgatgat gatgatgcac tttgtgattc atggtgtatt acctcttaat
4861 aattaagtgt gcctttcggc gatatggcgt gctttacaga ttttgaagcg ttaaaggaat
4921 gtgcactacg aggggtatca acgataactc ttgaagggac ttgccttact acactggata
4981 tgcgctagca catcaaattg ttatccgctc acaatttgat gtgcattaag ccacgcattg
5041 gcgggtgatg caacaattat ttttcatatt tatgattaat cggccacacc attcctt
//