质粒序列
LOCUS Exported 4405 bp ds-DNA circular SYN 29-七月-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pTrcHis2C
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4405)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2016年7月29日 from SnapGene Viewer 2.8.1
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..4405
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 193..222
/note="trc promoter"
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 230..246
/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)."
misc_feature 263..332
/note="rrnG antiterminator"
/note="antiterminator from the E. coli rrnG leader region
(Berg et al., 1989)"
CDS 383..409
/codon_start=1
/product="synthetic cistron containing a ribosome binding
site (Shine-Dalgarno sequence), for enhancing the bacterial
expression of a downstream cistron (Schoner, 1997)"
/note="minicistron"
/note="This first cistron should terminate 3 bp upstream of
the ATG for the second cistron."
/translation="MYRLNKEE"
misc_feature 411..543
/note="MCS"
CDS 470..499
/codon_start=1
/product="Myc (human c-Myc oncogene) epitope tag"
/note="Myc"
/translation="EQKLISEEDL"
CDS 515..532
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
terminator 758..844
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 936..963
/note="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 983..1074
/gene="bla"
/note="AmpR promoter"
CDS 1075..1935
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 2106..2694
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 2880..3020
/note="bom"
/note="basis of mobility region from pBR322"
promoter 3206..3283
/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 3284..4366
/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
YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
ORIGIN
1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
241 taacaatttc acacaggaaa cagcgccgct gagaaaaagc gaagcggcac tgctctttaa
301 caatttatca gacaatctgt gtgggcactc gaccggaatt atcgattaac tttattatta
361 aaaattaaag aggtatatat taatgtatcg attaaataag gaggaataaa ccatggatcc
421 gagctcgaga tctgcagctg gtaccatatg ggaattcgaa gcttacgtag aacaaaaact
481 catctcagaa gaggatctga atagcgccgt cgaccatcat catcatcatc attgagttta
541 aacggtctcc agcttggctg ttttggcgga tgagagaaga ttttcagcct gatacagatt
601 aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag tagcgcggtg
661 gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg
721 gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc
781 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
841 aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg
901 acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga cggatggcct
961 ttttgcgttt ctacaaactc tttttgttta tttttctaaa tacattcaaa tatgtatccg
1021 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
1081 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
1141 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
1201 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
1261 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
1321 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
1381 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
1441 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
1501 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
1561 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
1621 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
1681 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
1741 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
1801 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
1861 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
1921 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
1981 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
2041 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
2101 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
2161 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
2221 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
2281 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
2341 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
2401 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
2461 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
2521 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
2581 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
2641 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
2701 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
2761 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
2821 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
2881 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
2941 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
3001 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
3061 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
3121 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gcagatcaat tcgcgcgcga
3181 aggcgaagcg gcatgcattt acgttgacac catcgaatgg tgcaaaacct ttcgcggtat
3241 ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt
3301 atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca
3361 ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa
3421 ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt
3481 tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg
3541 cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc
3601 ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta
3661 tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt
3721 atttcttgat gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg
3781 tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc
3841 gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac
3901 tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt
3961 tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa
4021 cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc
4081 ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtc
4141 aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca
4201 actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag
4261 aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
4321 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
4381 atgtgagtta gcgcgaattg atctg
//