LOCUS Exported 5926 bp ds-DNA circular SYN 23-NOV-2017
DEFINITION synthetic circular DNA
KEYWORDS pET-49b
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5926)
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..5926
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 26..73
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(168..212)
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/label=S-Tag
/translation="KETAAAKFERQHMDS"
CDS complement(213..230)
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
CDS complement(309..332)
/codon_start=1
/product="recognition and cleavage site for human
rhinovirus 3C and PreScission proteases"
/label=HRV 3C site
/translation="LEVLFQGP"
CDS complement(342..359)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS complement(435..1088)
/codon_start=1
/product="glutathione S-transferase from Schistosoma
japonicum"
/label=GST
/translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
protein_bind 1133..1157
/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)."
promoter complement(1158..1176)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 1489..1566
/gene="lacI"
/label=lacI promoter
CDS 1567..2649
/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"
CDS 3221..3412
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
misc_feature 3514..3656
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(3842..4430)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 4552..5367
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin complement(5460..5915)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
ORIGIN
1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
121 tgtttagcag cctaggttaa ttaagcctcg agagcagcag aagtagagct gtccatgtgc
181 tggcgttcga atttagcagc agcggtttct ttactaccgc gtggcaccag agcgagctct
241 gcggccgcaa gcttgtcgac ggacgtcggg cgcgccaagg cctgtacaga attcggatcc
301 tggtacccgg gtccctgaaa gaggacttca agagccgcgg agtgatggtg atggtgatga
361 ccagaaccac tagatggagt aggagtagga ggattgttat tagaaccgcc accaccacta
421 gttgaaccat ccgattttgg aggatggtcg ccaccaccaa acgtggcttg ccagccctgc
481 aaaggccatg ctatatactt gctggatttc aagtacttat caatttgtgg gatagcttca
541 atacgttttt taaaacaaac taattttggg aacgcatcca ggcacattgg gtccatgtat
601 aaaacaacat caagagcgtc atacaacatg aagtcaggat gggttacatg atcaccattt
661 aaatatgttt tatgacataa acgatcttcg aacattttca gcatttcagg tagcttgcta
721 agaaaatcaa ctttgagagt ttcaaagtct ttactatatg caattctcga aacaccgtat
781 ctaatatcca aaaccgctcc ttcaagcatt gaaatctctg cacgctcttt tggacaacca
841 cccaacatgt tgtgcttgtc agctatataa cgtatgatgg ccatagactg tgttaattta
901 acatcaccat caatataata aggaagattg ggaaactcca aacccaattc aaactttttg
961 tttcgccatt tatcaccttc atcgcgctca tacaaatgct cttcatattt ttcttcaaga
1021 tattccaaaa gaagtcgagt gggttgcaca aggcccttaa ttttccaata acctagtata
1081 ggggacatat gtatatctcc ttcttaaagt taaacaaaat tatttctaga ggggaattgt
1141 tatccgctca caattcccct atagtgagtc gtattaattt cgcgggatcg agatcgatct
1201 cgatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg
1261 gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga
1321 gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg ttgggcgcca
1381 tctccttgca tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac ctactactgg
1441 gctgcttcct aatgcaggag tcgcataagg gagagcgtcg agatcccgga caccatcgaa
1501 tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg
1561 gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag
1621 accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg
1681 gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc
1741 aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa
1801 attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg
1861 gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc
1921 gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct
1981 gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt
2041 attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt
2101 caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg
2161 gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc
2221 gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt
2281 cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc
2341 gagtccgggc tgcgcgttgg tgcggacatc tcggtagtgg gatacgacga taccgaagac
2401 agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa
2461 accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg
2521 ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca aaccgcctct
2581 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
2641 gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact cattaggcac cgggatctcg
2701 accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac
2761 tagcatgatc gtgctcctgt cgttgaggac ccggctaggc tggcggggtt gccttactgg
2821 ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc
2881 tgcgacctga gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac
2941 gcggaagtca gcgccctgca ccattatgtt ccggatctgc atcgcaggat gctgctggct
3001 accctgtgga acacctacat ctgtattaac gaagcgctgg cattgaccct gagtgatttt
3061 tctctggtcc cgccgcatcc ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg
3121 ggcatgttca tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca tcggtatcat
3181 tacccccatg aacagaaatc ccccttacac ggaggcatca gtgaccaaac aggaaaaaac
3241 cgcccttaac atggcccgct ttatcagaag ccagacatta acgcttctgg agaaactcaa
3301 cgagctggac gcggatgaac aggcagacat ctgtgaatcg cttcacgacc acgctgatga
3361 gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
3421 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
3481 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga
3541 tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac
3601 catatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
3661 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
3721 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
3781 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
3841 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
3901 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
3961 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
4021 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
4081 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
4141 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
4201 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
4261 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
4321 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
4381 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
4441 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
4501 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat
4561 attcaacggg aaacgtcttg ctctaggccg cgattaaatt ccaacatgga tgctgattta
4621 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgattg
4681 tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat
4741 gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc
4801 atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccggc
4861 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg
4921 ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagt
4981 gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg
5041 agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat
5101 aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac
5161 cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca
5221 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta
5281 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt
5341 catttgatgc tcgatgagtt tttctaagaa ttaattcatg agcggataca tatttgaatg
5401 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
5461 aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
5521 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
5581 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
5641 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
5701 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
5761 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
5821 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
5881 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgcca
//