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载体序列
LOCUS pQLinkG2 5537 bp DNA circular SYN 12-JUL-2023
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5537)
AUTHORS Biofeng
TITLE Direct Submission
JOURNAL Exported Jul 12, 2023 from SnapGene 6.0.2
https://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5537
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 36..80
/label=T5 promoter
/note="bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator"
protein_bind 56..72
/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)."
protein_bind 88..104
/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)."
RBS 123..134
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 141..794
/codon_start=1
/product="glutathione S-transferase from Schistosoma
japonicum"
/label=GST
/translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
protein_bind 816..840
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 840..860
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
protein_bind complement(898..922)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
terminator 944..1038
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS 1132..1791
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
terminator 1856..1942
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(2035..3117)
/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(3118..3195)
/gene="lacI (mutant)"
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
misc_feature 3387..3527
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(3713..4301)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4472..5332)
/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(5333..5437)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ctcgagctta attaacaaca ccatttgtcg agaaatcata aaaaatttat ttgctttgtg
61 agcggataac aattataata gattcaattg tgagcggata acaatttcac acagaattca
121 ttaaagagga gaaattaact atgtccccta tactaggtta ttggaaaatt aagggccttg
181 tgcaacccac tcgacttctt ttggaatatc ttgaagaaaa atatgaagag catttgtatg
241 agcgcgatga aggtgataaa tggcgaaaca aaaagtttga attgggtttg gagtttccca
301 atcttcctta ttatattgat ggtgatgtta aattaacaca gtctatggcc atcatacgtt
361 atatagctga caagcacaac atgttgggtg gttgtccaaa agagcgtgca gagatttcaa
421 tgcttgaagg agcggttttg gatattagat acggtgtttc gagaattgca tatagtaaag
481 actttgaaac tctcaaagtt gattttctta gcaagctacc tgaaatgctg aaaatgttcg
541 aagatcgttt atgtcataaa acatatttga atggtgatca tgtaacccat cctgacttca
601 tgttgtatga cgctcttgat gttgttttat acatggaccc aatgtgcctg gatgcgttcc
661 caaaattagt ttgttttaaa aaacgtattg aagctatccc acaaattgat aagtacttga
721 aatccagcaa gtatatagca tggcctttgc agggctggca agccacgttt ggtggtggcg
781 accatcctcc aaaatcggat ctgggtgctg caggtacaag tttgtacaaa aaagcaggcg
841 agaatcttta ttttcaggga tccagtcttc gcatgcatga agacgcggcc gcctaggacc
901 cagctttctt gtacaaagtg gtaagcttaa ttagctgagc ttggactcct gttgatagat
961 ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg ccgccgggcg
1021 ttttttattg gtgagaatcc aagtaacaac accatttaaa tggagtggtt acaaatggag
1081 tggttaatta aggctagctt ggcgagattt tcaggagcta aggaagctaa aatggagaaa
1141 aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag
1201 gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc
1261 tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt
1321 gcccgcctga tgaatgctca tccggaattt cgtatggcaa tgaaagacgg tgagctggtg
1381 atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca
1441 tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat
1501 gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt
1561 ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg
1621 gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg
1681 ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga
1741 atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta atttttttaa
1801 ggcagttatt ggtgccctta aacgcctggg gtaatgactc tctagcttga ggcatcaaat
1861 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
1921 cgctctcctg agtaggacaa atccgccctc tagattacgt gcagtcgatg ataagctgtc
1981 aaacatgaga attgtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
2041 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
2101 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
2161 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
2221 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
2281 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga
2341 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
2401 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
2461 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
2521 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
2581 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
2641 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
2701 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
2761 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
2821 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
2881 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
2941 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
3001 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
3061 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
3121 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgca
3181 ccattcgatg gtgtcggaat ttcgggcagc gttgggtcct ggccacgggt gcgcatgatc
3241 tagagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
3301 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
3361 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
3421 gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccacatgc
3481 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt
3541 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
3601 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
3661 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
3721 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
3781 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
3841 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
3901 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
3961 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
4021 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
4081 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
4141 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
4201 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
4261 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
4321 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
4381 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
4441 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
4501 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
4561 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
4621 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
4681 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
4741 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
4801 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
4861 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
4921 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
4981 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
5041 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
5101 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
5161 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
5221 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
5281 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
5341 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
5401 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
5461 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
5521 ggccctttcg tcttcac
//
