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LOCUS pQLinkH 4901 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 4901)
AUTHORS Biofeng
TITLE Direct Submission
JOURNAL Exported Jul 12, 2023 from SnapGene 6.0.2
https://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4901
/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 147..167
/codon_start=1
/product="6xHis affinity tag"
/label=7xHis
/translation="HHHHHHH"
protein_bind 180..204
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 204..224
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
protein_bind complement(262..286)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
terminator 308..402
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS 496..1155
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
terminator 1220..1306
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(1399..2481)
/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(2482..2559)
/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 2751..2891
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(3077..3665)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3836..4696)
/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(4697..4801)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ctcgagctta attaacaaca ccatttgtcg agaaatcata aaaaatttat ttgctttgtg
61 agcggataac aattataata gattcaattg tgagcggata acaatttcac acagaattca
121 ttaaagagga gaaattaact atgaaacatc accatcacca tcaccatggt gctgcaggta
181 caagtttgta caaaaaagca ggcgagaatc tttattttca gggatccagt cttcgcatgc
241 atgaagacgc ggccgcctag gacccagctt tcttgtacaa agtggtaagc ttaattagct
301 gagcttggac tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc
361 agaacgctcg gttgccgccg ggcgtttttt attggtgaga atccaagtaa caacaccatt
421 taaatggagt ggttacaaat ggagtggtta attaaggcta gcttggcgag attttcagga
481 gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa
541 tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag
601 accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt
661 tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga atttcgtatg
721 gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc
781 catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag
841 tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct
901 aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt
961 tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa
1021 tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt
1081 tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg
1141 cagggcgggg cgtaattttt ttaaggcagt tattggtgcc cttaaacgcc tggggtaatg
1201 actctctagc ttgaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
1261 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cctctagatt
1321 acgtgcagtc gatgataagc tgtcaaacat gagaattgtg cctaatgagt gagctaactt
1381 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
1441 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt
1501 ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga
1561 gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt
1621 ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat
1681 atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg
1741 atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg
1801 ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt
1861 gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg
1921 gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg
1981 cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag
2041 aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag
2101 cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt
2161 acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc
2221 ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt
2281 ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat
2341 gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct
2401 ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc
2461 gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca
2521 tgccataccg cgaaaggttt tgcaccattc gatggtgtcg gaatttcggg cagcgttggg
2581 tcctggccac gggtgcgcat gatctagagc tgcctcgcgc gtttcggtga tgacggtgaa
2641 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
2701 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg
2761 acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga
2821 ttgtactgag agtgcaccac atgcggtgtg aaataccgca cagatgcgta aggagaaaat
2881 accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
2941 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
3001 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
3061 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
3121 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
3181 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
3241 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
3301 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
3361 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
3421 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
3481 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
3541 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
3601 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
3661 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
3721 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
3781 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
3841 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
3901 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
3961 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
4021 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
4081 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
4141 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
4201 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
4261 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
4321 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
4381 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
4441 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
4501 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
4561 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
4621 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
4681 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
4741 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
4801 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
4861 cctataaaaa taggcgtatc acgaggccct ttcgtcttca c
//
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