LOCUS Exported File 9004 bp ds-DNA circular SYN 31-1-2015
DEFINITION .
ACCESSION .
VERSION .
KEYWORDS Untitled
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9004)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2015-1-31 from MLCC
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..9004
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 1049..1295
/gene="Pichia pastoris AOX1"
/note="AOX1 terminator"
/note="transcription terminator for AOX1"
CDS complement(1708..4242)
/codon_start=1
/gene="Pichia pastoris HIS4"
/locus_tag="
"
/product="multifunctional enzyme, required for histidine
biosynthesis"
/note="PpHIS4"
/note="auxotrophic marker for Pichia pastoris"
/protein_id="
"
/translation="MTFPLLPAYASVAEFDNSLSLVGKAVFPYAADQLHNLIKFTQSTE
LQVNVQVESSVTEDQFEELIDNLLKLYNNGINEVILDLDLAERVVQRMIPGARVIYRTL
VDKVASLPANASIAVPFSSPLGDLKSFTNGGSRTVYAFSETAKLVDVTSTVASGIIPII
DARQLTTEYELSEDVKKFPVSEILLASLTTDRPDGLFTTLVADSSNYSLGLVYSSKKSI
PEAIRTQTGVYQSRRHGLWYKGATSGATQKLLGIELDCDGDCLKFVVEQTGVGFCHLER
TSCFGQSKGLRAMEATLWDRKSNAPEGSYTKRLFDDEVLLNAKIREEADELAEAKSKED
IAWECADLFYFALVRCAKYGVTLDEVERNLDMKSLKVTRRKGDAKPGYTKEQPKEESKP
KEVPSEGRIELCKIDVSKASSQEIEDALRRPIQKTEQIMELVKPIVDNVRQNGDKALLE
LTAKFDGVALKTPVLEAPFPEELMQLPDNVKRAIDLSIDNVRKFHEAQLTETLQVETCP
GVVCSRFARPIEKVGLYIPGGTAILPSTSLMLGVPAKVAGCKEIVFASPPKKDGTLTPE
VIYVAHKVGAKCIVLAGGAQAVAAMAYGTETVPKCDKIFGPGNQFVTAAKMMVQNDTSA
LCSIDMPAGPSEVLVIADKYADPDFVASDLLSQAEHGIDSQVILLAVDMTDKELARIED
AVHNQAVQLPRVEIVRKCIAHSTTLSVATYEQALEMSNQYAPEHLILQIENASSYVDQV
QHAGSVFVGAYSPESCGDYSSGTNHTLPTYGYARQYSGVNTATFQKFITSQDVTPEGLK
HIGQAVMDLAAVEGLDAHRNAVKVRMEKLGLI"
CDS complement(4656..5471)
/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 5850..6606
/note="AOX1 3' fragment"
/note="region downstream of Pichia pastoris AOX1 gene"
misc_feature 6749..6889
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(7075..7663)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7834..8694)
/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(8695..8799)
/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 tcctacgtag aattccctag
961 ggcggccgcg aattaattcg ccttagacat gactgttcct cagttcaagt tgggcactta
1021 cgagaagacc ggtcttgcta gattctaatc aagaggatgt cagaatgcca tttgcctgag
1081 agatgcaggc ttcatttttg atactttttt atttgtaacc tatatagtat aggatttttt
1141 ttgtcatttt gtttcttctc gtacgagctt gctcctgatc agcctatctc gcagctgatg
1201 aatatcttgt ggtaggggtt tgggaaaatc attcgagttt gatgtttttc ttggtatttc
1261 ccactcctct tcagagtaca gaagattaag tgagaagttc gtttgtgcaa gcttatcgat
1321 aagctttaat gcggtagttt atcacagtta aattgctaac gcagtcaggc accgtgtatg
1381 aaatctaaca atgcgctcat cgtcatcctc ggcaccgtca ccctggatgc tgtaggcata
1441 ggcttggtta tgccggtact gccgggcctc ttgcgggata tcgtccattc cgacagcatc
1501 gccagtcact atggcgtgct gctagcgcta tatgcgttga tgcaatttct atgcgcaccc
1561 gttctcggag cactgtccga ccgctttggc cgccgcccag tcctgctcgc ttcgctactt
1621 ggagccacta tcgactacgc gatcatggcg accacacccg tcctgtggat ctatcgaatc
1681 taaatgtaag ttaaaatctc taaataatta aataagtccc agtttctcca tacgaacctt
1741 aacagcattg cggtgagcat ctagaccttc aacagcagcc agatccatca ctgcttggcc
1801 aatatgtttc agtccctcag gagttacgtc ttgtgaagtg atgaacttct ggaaggttgc
1861 agtgttaact ccgctgtatt gacgggcata tccgtacgtt ggcaaagtgt ggttggtacc
1921 ggaggagtaa tctccacaac tctctggaga gtaggcacca acaaacacag atccagcgtg
1981 ttgtacttga tcaacataag aagaagcatt ctcgatttgc aggatcaagt gttcaggagc
2041 gtactgattg gacatttcca aagcctgctc gtaggttgca accgataggg ttgtagagtg
2101 tgcaatacac ttgcgtacaa tttcaaccct tggcaactgc acagcttggt tgtgaacagc
2161 atcttcaatt ctggcaagct ccttgtctgt catatcgaca gccaacagaa tcacctggga
2221 atcaatacca tgttcagctt gagacagaag gtctgaggca acgaaatctg gatcagcgta
2281 tttatcagca ataactagaa cttcagaagg cccagcaggc atgtcaatac tacacagggc
2341 tgatgtgtca ttttgaacca tcatcttggc agcagtaacg aactggtttc ctggaccaaa
2401 tattttgtca cacttaggaa cagtttctgt tccgtaagcc atagcagcta ctgcctgggc
2461 gcctcctgct agcacgatac acttagcacc aaccttgtgg gcaacgtaga tgacttctgg
2521 ggtaagggta ccatccttct taggtggaga tgcaaaaaca atttctttgc aaccagcaac
2581 tttggcagga acacccagca tcagggaagt ggaaggcaga attgcggttc caccaggaat
2641 atagaggcca actttctcaa taggtcttgc aaaacgagag cagactacac cagggcaagt
2701 ctcaacttgc aacgtctccg ttagttgagc ttcatggaat ttcctgacgt tatctataga
2761 gagatcaatg gctctcttaa cgttatctgg caattgcata agttcctctg ggaaaggagc
2821 ttctaacaca ggtgtcttca aagcgactcc atcaaacttg gcagttagtt ctaaaagggc
2881 tttgtcacca ttttgacgaa cattgtcgac aattggtttg actaattcca taatctgttc
2941 cgttttctgg ataggacgac gaagggcatc ttcaatttct tgtgaggagg ccttagaaac
3001 gtcaattttg cacaattcaa tacgaccttc agaagggact tctttaggtt tggattcttc
3061 tttaggttgt tccttggtgt atcctggctt ggcatctcct ttccttctag tgacctttag
3121 ggacttcata tccaggtttc tctccacctc gtccaacgtc acaccgtact tggcacatct
3181 aactaatgca aaataaaata agtcagcaca ttcccaggct atatcttcct tggatttagc
3241 ttctgcaagt tcatcagctt cctccctaat tttagcgttc aacaaaactt cgtcgtcaaa
3301 taaccgtttg gtataagaac cttctggagc attgctctta cgatcccaca aggtggcttc
3361 catggctcta agaccctttg attggccaaa acaggaagtg cgttccaagt gacagaaacc
3421 aacacctgtt tgttcaacca caaatttcaa gcagtctcca tcacaatcca attcgatacc
3481 cagcaacttt tgagttgctc cagatgtagc acctttatac cacaaaccgt gacgacgaga
3541 ttggtagact ccagtttgtg tccttatagc ctccggaata gactttttgg acgagtacac
3601 caggcccaac gagtaattag aagagtcagc caccaaagta gtgaatagac catcggggcg
3661 gtcagtagtc aaagacgcca acaaaatttc actgacaggg aactttttga catcttcaga
3721 aagttcgtat tcagtagtca attgccgagc atcaataatg gggattatac cagaagcaac
3781 agtggaagtc acatctacca actttgcggt ctcagaaaaa gcataaacag ttctactacc
3841 gccattagtg aaacttttca aatcgcccag tggagaagaa aaaggcacag cgatactagc
3901 attagcgggc aaggatgcaa ctttatcaac cagggtccta tagataaccc tagcgcctgg
3961 gatcatcctt tggacaactc tttctgccaa atctaggtcc aaaatcactt cattgatacc
4021 attattgtac aacttgagca agttgtcgat cagctcctca aattggtcct ctgtaacgga
4081 tgactcaact tgcacattaa cttgaagctc agtcgattga gtgaacttga tcaggttgtg
4141 cagctggtca gcagcatagg gaaacacggc ttttcctacc aaactcaagg aattatcaaa
4201 ctctgcaaca cttgcgtatg caggtagcaa gggaaatgtc atacttgaag tcggacagtg
4261 agtgtagtct tgagaaattc tgaagccgta tttttattat cagtgagtca gtcatcagga
4321 gatcctctac gccggacgca tcgtggccga cctgcagggg gggggggggc gctgaggtct
4381 gcctcgtgaa gaaggtgttg ctgactcata ccaggcctga atcgccccat catccagcca
4441 gaaagtgagg gagccacggt tgatgagagc tttgttgtag gtggaccagt tggtgatttt
4501 gaacttttgc tttgccacgg aacggtctgc gttgtcggga agatgcgtga tctgatcctt
4561 caactcagca aaagttcgat ttattcaaca aagccgccgt cccgtcaagt cagcgtaatg
4621 ctctgccagt gttacaacca attaaccaat tctgattaga aaaactcatc gagcatcaaa
4681 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
4741 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
4801 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
4861 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagc
4921 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
4981 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
5041 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
5101 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
5161 ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
5221 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
5281 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
5341 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
5401 tataaatcag catccatgtt ggaatttaat cgcggcctcg agcaagacgt ttcccgttga
5461 atatggctca taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat
5521 gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtggctt
5581 tccccccccc ccctgcaggt cggcatcacc ggcgccacag gtgcggttgc tggcgcctat
5641 atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcgcttgt
5701 ttcggcgtgg gtatggtggc aggccccgtg gccgggggac tgttgggcgc catctccttg
5761 catgcaccat tccttgcggc ggcggtgctc aacggcctca acctactact gggctgcttc
5821 ctaatgcagg agtcgcataa gggagagcgt cgagtatcta tgattggaag tatgggaatg
5881 gtgatacccg cattcttcag tgtcttgagg tctcctatca gattatgccc aactaaagca
5941 accggaggag gagatttcat ggtaaatttc tctgactttt ggtcatcagt agactcgaac
6001 tgtgagacta tctcggttat gacagcagaa atgtccttct tggagacagt aaatgaagtc
6061 ccaccaataa agaaatcctt gttatcagga acaaacttct tgtttcgaac tttttcggtg
6121 ccttgaacta taaaatgtag agtggatatg tcgggtagga atggagcggg caaatgctta
6181 ccttctggac cttcaagagg tatgtagggt ttgtagatac tgatgccaac ttcagtgaca
6241 acgttgctat ttcgttcaaa ccattccgaa tccagagaaa tcaaagttgt ttgtctacta
6301 ttgatccaag ccagtgcggt cttgaaactg acaatagtgt gctcgtgttt tgaggtcatc
6361 tttgtatgaa taaatctagt ctttgatcta aataatcttg acgagccaag gcgataaata
6421 cccaaatcta aaactctttt aaaacgttaa aaggacaagt atgtctgcct gtattaaacc
6481 ccaaatcagc tcgtagtctg atcctcatca acttgagggg cactatcttg ttttagagaa
6541 atttgcggag atgcgatatc gagaaaaagg tacgctgatt ttaaacgtga aatttatctc
6601 aagatctctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
6661 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
6721 cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg
6781 gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
6841 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc
6901 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
6961 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
7021 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
7081 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
7141 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
7201 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
7261 gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
7321 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
7381 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
7441 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
7501 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
7561 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
7621 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
7681 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
7741 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
7801 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
7861 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
7921 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
7981 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
8041 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
8101 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgctg caggcatcgt
8161 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
8221 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
8281 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
8341 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
8401 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa
8461 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
8521 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
8581 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
8641 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
8701 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
8761 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
8821 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
8881 gaggcccttt cgtcttcaag aattaattct catgtttgac agcttatcat cgataagctg
8941 actcatgttg gtattgtgaa atagacgcag atcgggaaca ctgaaaaata acagttatta
9001 ttcg
//
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