LOCUS Exported 9276 bp ds-DNA circular SYN 10-SEP-2016
DEFINITION synthetic circular DNA
KEYWORDS Untitled 2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9276)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..9276
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 2..937
/gene="Pichia pastoris AOX1"
/note="AOX1 promoter"
/note="inducible promoter, regulated by methanol"
CDS 949..1215
/codon_start=1
/gene="MF-alpha-1"
/product="N-terminal secretion signal from S. cerevisiae
alpha-factor"
/note="alpha-factor secretion signal"
/note="Cleavage by the Kex2 protease occurs after the
dibasic KR sequence. The EA dipeptides are then removed by
dipeptidyl aminopeptidase A."
/translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA"
terminator 1321..1567
/gene="Pichia pastoris AOX1"
/note="AOX1 terminator"
/note="transcription terminator for AOX1"
CDS complement(1980..4514)
/codon_start=1
/gene="Pichia pastoris HIS4"
/product="multifunctional enzyme, required for histidine
biosynthesis"
/note="PpHIS4"
/note="auxotrophic marker for Pichia pastoris"
/translation="MTFPLLPAYASVAEFDNSLSLVGKAVFPYAADQLHNLIKFTQSTE
LQVNVQVESSVTEDQFEELIDNLLKLYNNGINEVILDLDLAERVVQRMIPGARVIYRTL
VDKVASLPANASIAVPFSSPLGDLKSFTNGGSRTVYAFSETAKLVDVTSTVASGIIPII
DARQLTTEYELSEDVKKFPVSEILLASLTTDRPDGLFTTLVADSSNYSLGLVYSSKKSI
PEAIRTQTGVYQSRRHGLWYKGATSGATQKLLGIELDCDGDCLKFVVEQTGVGFCHLER
TSCFGQSKGLRAMEATLWDRKSNAPEGSYTKRLFDDEVLLNAKIREEADELAEAKSKED
IAWECADLFYFALVRCAKYGVTLDEVERNLDMKSLKVTRRKGDAKPGYTKEQPKEESKP
KEVPSEGRIELCKIDVSKASSQEIEDALRRPIQKTEQIMELVKPIVDNVRQNGDKALLE
LTAKFDGVALKTPVLEAPFPEELMQLPDNVKRAIDLSIDNVRKFHEAQLTETLQVETCP
GVVCSRFARPIEKVGLYIPGGTAILPSTSLMLGVPAKVAGCKEIVFASPPKKDGTLTPE
VIYVAHKVGAKCIVLAGGAQAVAAMAYGTETVPKCDKIFGPGNQFVTAAKMMVQNDTSA
LCSIDMPAGPSEVLVIADKYADPDFVASDLLSQAEHGIDSQVILLAVDMTDKELARIED
AVHNQAVQLPRVEIVRKCIAHSTTLSVATYEQALEMSNQYAPEHLILQIENASSYVDQV
QHAGSVFVGAYSPESCGDYSSGTNHTLPTYGYARQYSGVNTATFQKFITSQDVTPEGLK
HIGQAVMDLAAVEGLDAHRNAVKVRMEKLGLI"
CDS complement(4928..5743)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/note="KanR"
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
misc_feature 6122..6878
/note="AOX1 3' fragment"
/note="region downstream of Pichia pastoris AOX1 gene"
misc_feature 7021..7161
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(7347..7935)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(8106..8966)
/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"
promoter complement(8967..9071)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 agatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag
61 gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt
121 tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc
181 agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta
241 acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta
301 tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg
361 agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct
421 gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg
481 ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcgcca taccgtttgt
541 cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct
601 ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct
661 ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact
721 gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat
781 atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt
841 actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga
901 caacttgaga agatcaaaaa acaactaatt attcgaagga tccaaacgat gagatttcct
961 tcaattttta ctgcagtttt attcgcagca tcctccgcat tagctgctcc agtcaacact
1021 acaacagaag atgaaacggc acaaattccg gctgaagctg tcatcggtta ctcagattta
1081 gaaggggatt tcgatgttgc tgttttgcca ttttccaaca gcacaaataa cgggttattg
1141 tttataaata ctactattgc cagcattgct gctaaagaag aaggggtatc tctcgagaaa
1201 agagaggctg aagcttacgt agaattccct agggcggccg cgaattaatt cgccttagac
1261 atgactgttc ctcagttcaa gttgggcact tacgagaaga ccggtcttgc tagattctaa
1321 tcaagaggat gtcagaatgc catttgcctg agagatgcag gcttcatttt tgatactttt
1381 ttatttgtaa cctatatagt ataggatttt ttttgtcatt ttgtttcttc tcgtacgagc
1441 ttgctcctga tcagcctatc tcgcagctga tgaatatctt gtggtagggg tttgggaaaa
1501 tcattcgagt ttgatgtttt tcttggtatt tcccactcct cttcagagta cagaagatta
1561 agtgagaagt tcgtttgtgc aagcttatcg ataagcttta atgcggtagt ttatcacagt
1621 taaattgcta acgcagtcag gcaccgtgta tgaaatctaa caatgcgctc atcgtcatcc
1681 tcggcaccgt caccctggat gctgtaggca taggcttggt tatgccggta ctgccgggcc
1741 tcttgcggga tatcgtccat tccgacagca tcgccagtca ctatggcgtg ctgctagcgc
1801 tatatgcgtt gatgcaattt ctatgcgcac ccgttctcgg agcactgtcc gaccgctttg
1861 gccgccgccc agtcctgctc gcttcgctac ttggagccac tatcgactac gcgatcatgg
1921 cgaccacacc cgtcctgtgg atctatcgaa tctaaatgta agttaaaatc tctaaataat
1981 taaataagtc ccagtttctc catacgaacc ttaacagcat tgcggtgagc atctagacct
2041 tcaacagcag ccagatccat cactgcttgg ccaatatgtt tcagtccctc aggagttacg
2101 tcttgtgaag tgatgaactt ctggaaggtt gcagtgttaa ctccgctgta ttgacgggca
2161 tatccgtacg ttggcaaagt gtggttggta ccggaggagt aatctccaca actctctgga
2221 gagtaggcac caacaaacac agatccagcg tgttgtactt gatcaacata agaagaagca
2281 ttctcgattt gcaggatcaa gtgttcagga gcgtactgat tggacatttc caaagcctgc
2341 tcgtaggttg caaccgatag ggttgtagag tgtgcaatac acttgcgtac aatttcaacc
2401 cttggcaact gcacagcttg gttgtgaaca gcatcttcaa ttctggcaag ctccttgtct
2461 gtcatatcga cagccaacag aatcacctgg gaatcaatac catgttcagc ttgagacaga
2521 aggtctgagg caacgaaatc tggatcagcg tatttatcag caataactag aacttcagaa
2581 ggcccagcag gcatgtcaat actacacagg gctgatgtgt cattttgaac catcatcttg
2641 gcagcagtaa cgaactggtt tcctggacca aatattttgt cacacttagg aacagtttct
2701 gttccgtaag ccatagcagc tactgcctgg gcgcctcctg ctagcacgat acacttagca
2761 ccaaccttgt gggcaacgta gatgacttct ggggtaaggg taccatcctt cttaggtgga
2821 gatgcaaaaa caatttcttt gcaaccagca actttggcag gaacacccag catcagggaa
2881 gtggaaggca gaattgcggt tccaccagga atatagaggc caactttctc aataggtctt
2941 gcaaaacgag agcagactac accagggcaa gtctcaactt gcaacgtctc cgttagttga
3001 gcttcatgga atttcctgac gttatctata gagagatcaa tggctctctt aacgttatct
3061 ggcaattgca taagttcctc tgggaaagga gcttctaaca caggtgtctt caaagcgact
3121 ccatcaaact tggcagttag ttctaaaagg gctttgtcac cattttgacg aacattgtcg
3181 acaattggtt tgactaattc cataatctgt tccgttttct ggataggacg acgaagggca
3241 tcttcaattt cttgtgagga ggccttagaa acgtcaattt tgcacaattc aatacgacct
3301 tcagaaggga cttctttagg tttggattct tctttaggtt gttccttggt gtatcctggc
3361 ttggcatctc ctttccttct agtgaccttt agggacttca tatccaggtt tctctccacc
3421 tcgtccaacg tcacaccgta cttggcacat ctaactaatg caaaataaaa taagtcagca
3481 cattcccagg ctatatcttc cttggattta gcttctgcaa gttcatcagc ttcctcccta
3541 attttagcgt tcaacaaaac ttcgtcgtca aataaccgtt tggtataaga accttctgga
3601 gcattgctct tacgatccca caaggtggct tccatggctc taagaccctt tgattggcca
3661 aaacaggaag tgcgttccaa gtgacagaaa ccaacacctg tttgttcaac cacaaatttc
3721 aagcagtctc catcacaatc caattcgata cccagcaact tttgagttgc tccagatgta
3781 gcacctttat accacaaacc gtgacgacga gattggtaga ctccagtttg tgtccttata
3841 gcctccggaa tagacttttt ggacgagtac accaggccca acgagtaatt agaagagtca
3901 gccaccaaag tagtgaatag accatcgggg cggtcagtag tcaaagacgc caacaaaatt
3961 tcactgacag ggaacttttt gacatcttca gaaagttcgt attcagtagt caattgccga
4021 gcatcaataa tggggattat accagaagca acagtggaag tcacatctac caactttgcg
4081 gtctcagaaa aagcataaac agttctacta ccgccattag tgaaactttt caaatcgccc
4141 agtggagaag aaaaaggcac agcgatacta gcattagcgg gcaaggatgc aactttatca
4201 accagggtcc tatagataac cctagcgcct gggatcatcc tttggacaac tctttctgcc
4261 aaatctaggt ccaaaatcac ttcattgata ccattattgt acaacttgag caagttgtcg
4321 atcagctcct caaattggtc ctctgtaacg gatgactcaa cttgcacatt aacttgaagc
4381 tcagtcgatt gagtgaactt gatcaggttg tgcagctggt cagcagcata gggaaacacg
4441 gcttttccta ccaaactcaa ggaattatca aactctgcaa cacttgcgta tgcaggtagc
4501 aagggaaatg tcatacttga agtcggacag tgagtgtagt cttgagaaat tctgaagccg
4561 tatttttatt atcagtgagt cagtcatcag gagatcctct acgccggacg catcgtggcc
4621 gacctgcagg gggggggggg gcgctgaggt ctgcctcgtg aagaaggtgt tgctgactca
4681 taccaggcct gaatcgcccc atcatccagc cagaaagtga gggagccacg gttgatgaga
4741 gctttgttgt aggtggacca gttggtgatt ttgaactttt gctttgccac ggaacggtct
4801 gcgttgtcgg gaagatgcgt gatctgatcc ttcaactcag caaaagttcg atttattcaa
4861 caaagccgcc gtcccgtcaa gtcagcgtaa tgctctgcca gtgttacaac caattaacca
4921 attctgatta gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat
4981 tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc
5041 agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa
5101 tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag
5161 tgacgactga atccggtgag aatggcaaaa gcttatgcat ttctttccag acttgttcaa
5221 caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc
5281 gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag
5341 gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat
5401 caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc
5461 atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca
5521 gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt
5581 tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt
5641 gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta
5701 atcgcggcct cgagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac
5761 tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt
5821 aacatcagag attttgagac acaacgtggc tttccccccc ccccctgcag gtcggcatca
5881 ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc
5941 gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg
6001 tggccggggg actgttgggc gccatctcct tgcatgcacc attccttgcg gcggcggtgc
6061 tcaacggcct caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc
6121 gtcgagtatc tatgattgga agtatgggaa tggtgatacc cgcattcttc agtgtcttga
6181 ggtctcctat cagattatgc ccaactaaag caaccggagg aggagatttc atggtaaatt
6241 tctctgactt ttggtcatca gtagactcga actgtgagac tatctcggtt atgacagcag
6301 aaatgtcctt cttggagaca gtaaatgaag tcccaccaat aaagaaatcc ttgttatcag
6361 gaacaaactt cttgtttcga actttttcgg tgccttgaac tataaaatgt agagtggata
6421 tgtcgggtag gaatggagcg ggcaaatgct taccttctgg accttcaaga ggtatgtagg
6481 gtttgtagat actgatgcca acttcagtga caacgttgct atttcgttca aaccattccg
6541 aatccagaga aatcaaagtt gtttgtctac tattgatcca agccagtgcg gtcttgaaac
6601 tgacaatagt gtgctcgtgt tttgaggtca tctttgtatg aataaatcta gtctttgatc
6661 taaataatct tgacgagcca aggcgataaa tacccaaatc taaaactctt ttaaaacgtt
6721 aaaaggacaa gtatgtctgc ctgtattaaa ccccaaatca gctcgtagtc tgatcctcat
6781 caacttgagg ggcactatct tgttttagag aaatttgcgg agatgcgata tcgagaaaaa
6841 ggtacgctga ttttaaacgt gaaatttatc tcaagatctc tgcctcgcgc gtttcggtga
6901 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
6961 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
7021 cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca
7081 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
7141 aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
7201 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
7261 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
7321 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
7381 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
7441 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
7501 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
7561 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
7621 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
7681 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
7741 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
7801 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
7861 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
7921 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
7981 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
8041 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
8101 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
8161 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
8221 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
8281 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
8341 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
8401 cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct
8461 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
8521 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
8581 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
8641 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
8701 agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa
8761 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
8821 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
8881 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
8941 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
9001 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
9061 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
9121 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca agaattaatt
9181 ctcatgtttg acagcttatc atcgataagc tgactcatgt tggtattgtg aaatagacgc
9241 agatcgggaa cactgaaaaa taacagttat tattcg
//
Caution:
1. This product is FOR RESEARCH USE ONLY!
2. The item is lyophilized form, Please take the powder plasmid by centrifugation at 5000rpm/min for 1min. Add 20μl ddH2O in to the tube of plasmid.
Search name
pPIC9k,Plasmid pPIC9k,pPIC9k vector
English