质粒序列
LOCUS Exported File 4972 bp ds-DNA circular SYN 12-4-2015
DEFINITION .
ACCESSION .
VERSION .
KEYWORDS Untitled
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4972)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2015-4-12 from MLCC
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..4972
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 183..211
/note="tac promoter"
/note="strong E. coli promoter; hybrid between the trp and
lac?UV5 promoters"
protein_bind 219..235
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/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 258..911
/codon_start=1
/gene="
"
/product="glutathione S-transferase from Schistosoma
japonicum"
/note="GST"
/translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
CDS 921..932
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/note="Factor Xa site"
/translation="IEGR"
promoter 1275..1379
/gene="bla"
/note="AmpR promoter"
CDS 1380..2240
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 2411..2999
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 3243..3320
/gene="lacI (mutant)"
/note="lacIq promoter"
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
CDS 3321..4403
/codon_start=1
/gene="lacI"
/product="lac repressor"
/note="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"
protein_bind 4416..4437
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4452..4482
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 4490..4506
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/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 4514..4530
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4546..4562)
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 agcttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
901 atcctccaaa atcggatctg atcgaaggtc gtgggatccc cgaattcccg ggtcgactcg
961 agcggccgca tcgtgactga ctgacgatct gcctcgcgcg tttcggtgat gacggtgaaa
1021 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
1081 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga
1141 cccagtcacg tagcgatagc ggagtgtata attcttgaag acgaaagggc ctcgtgatac
1201 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
1261 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
1321 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
1381 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
1441 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
1501 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
1561 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
1621 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
1681 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
1741 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
1801 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
1861 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
1921 ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
1981 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
2041 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
2101 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
2161 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
2221 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
2281 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
2341 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
2401 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
2461 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
2521 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
2581 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
2641 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
2701 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
2761 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
2821 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
2881 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
2941 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
3001 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
3061 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
3121 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
3181 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcataaatt
3241 ccgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
3301 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
3361 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
3421 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
3481 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
3541 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
3601 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
3661 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
3721 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
3781 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
3841 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
3901 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
3961 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
4021 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
4081 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg
4141 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
4201 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
4261 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
4321 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
4381 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
4441 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
4501 taacaatttc acacaggaaa cagctatgac catgattacg gattcactgg ccgtcgtttt
4561 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
4621 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
4681 gcgcagcctg aatggcgaat ggcgctttgc ctggtttccg gcaccagaag cggtgccgga
4741 aagctggctg gagtgcgatc ttcctgaggc cgatactgtc gtcgtcccct caaactggca
4801 gatgcacggt tacgatgcgc ccatctacac caacgtaacc tatcccatta cggtcaatcc
4861 gccgtttgtt cccacggaga atccgacggg ttgttactcg ctcacattta atgttgatga
4921 aagctggcta caggaaggcc agacgcgaat tatttttgat ggcgttggaa tt
//