质粒序列
LOCUS Exported 3996 bp ds-DNA circular SYN 08-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 6
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3996)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Thursday, September 8, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..3996
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 10..54
/note="T5 promoter"
/note="bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator"
protein_bind 30..46
/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)."
protein_bind 62..78
/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)."
RBS 97..108
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 133..690
/codon_start=1
/gene="Mus musculus Dhfr"
/product="mouse dihydrofolate reductase"
/note="DHFR"
/translation="MVRPLNSIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
PGVLSEVQEEKGIKYKFEVYEKK"
CDS 703..720
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
terminator 743..837
/note="lambda t0 terminator"
/note="transcription terminator from phage lambda"
CDS 881..1540
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
terminator 1605..1691
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
misc_feature 1846..1986
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(2172..2760)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2931..3791)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(3792..3896)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga
121 ggatccggca tcatggttcg accattgaac tcgatcgtcg ccgtgtccca aaatatgggg
181 attggcaaga acggagacct accctggcct ccgctcagga acgagttcaa gtacttccaa
241 agaatgacca caacctcttc agtggaaggt aaacagaatc tggtgattat gggtaggaaa
301 acctggttct ccattcctga gaagaatcga cctttaaagg acagaattaa tatagttctc
361 agtagagaac tcaaagaacc accacgagga gctcattttc ttgccaaaag tttggatgat
421 gccttaagac ttattgaaca accggaattg gcaagtaaag tagacatggt ttggatagtc
481 ggaggcagtt ctgtttacca ggaagccatg aatcaaccag gccaccttag actctttgtg
541 acaaggatca tgcaggaatt tgaaagtgac acgtttttcc cagaaattga tttggggaaa
601 tataaacttc tcccagaata cccaggcgtc ctctctgagg tccaggagga aaaaggcatc
661 aagtataagt ttgaagtcta cgagaagaaa ggttccagat ctcatcacca tcaccatcac
721 taagcttaat tagctgagct tggactcctg ttgatagatc cagtaatgac ctcagaactc
781 catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg tgagaatcca
841 agctagcttg gcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg
901 atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc
961 agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac
1021 cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat
1081 gaatgctcat ccggaatttc gtatggcaat gaaagacggt gagctggtga tatgggatag
1141 tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag
1201 tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta
1261 cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc
1321 caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt
1381 cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct
1441 ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga
1501 attacaacag tactgcgatg agtggcaggg cggggcgtaa tttttttaag gcagttattg
1561 gtgcccttaa acgcctgggg taatgactct ctagcttgag gcatcaaata aaacgaaagg
1621 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
1681 gtaggacaaa tccgccctct agagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
1741 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
1801 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
1861 gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta
1921 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
1981 atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
2041 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
2101 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
2161 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
2221 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
2281 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
2341 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
2401 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
2461 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
2521 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
2581 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
2641 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
2701 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
2761 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
2821 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
2881 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
2941 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
3001 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
3061 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
3121 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
3181 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
3241 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
3301 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
3361 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
3421 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
3481 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
3541 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
3601 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
3661 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
3721 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
3781 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
3841 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
3901 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
3961 aaaaataggc gtatcacgag gccctttcgt cttcac
//