LOCUS Exported 6018 bp ds-DNA circular SYN 12-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 5
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6018)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Monday, September 12, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..6018
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(663..1757)
/codon_start=1
/gene="S. cerevisiae LEU2"
/product="3-isopropylmalate dehydrogenase, required for
leucine biosynthesis"
/note="LEU2"
/note="yeast auxotrophic marker"
/translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
DLGGSNSTTEVGDAVAEEVKKILA"
promoter complement(1770..2174)
/gene="S. cerevisiae LEU2"
/note="LEU2 promoter"
rep_origin complement(2474..2929)
/direction=LEFT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(2712..3287)
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/note="lacZ-alpha"
/translation="MTMITPSSELTLTKGNKSWVPGPPSRSTVSISLISNSCSPGDPLV
LERPPPRWSSNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
DRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFAG
FPRQALNRGLPLGFRFSALRHLDPKKLD"
primer_bind 3074..3090
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter 3097..3115
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 3124..3231
/note="MCS"
/note="pBluescript multiple cloning site"
primer_bind 3148..3164
/note="SK primer"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3198..3214)
/note="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(3244..3262)
/note="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(3283..3299)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 3307..3323
/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)."
promoter complement(3331..3361)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
rep_origin complement(3685..4273)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4444..5304)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5305..5409)
/gene="bla"
/note="AmpR promoter"
misc_feature 5446..5949
/note="CEN/ARS"
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
ORIGIN
1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
181 accatatcga ctacgtcgta aggccgtttc tgacagagta aaattcttga gggaactttc
241 accattatgg gaaatggttc aagaaggtat tgacttaaac tccatcaaat ggtcaggtca
301 ttgagtgttt tttatttgtt gtattttttt ttttttagag aaaatcctcc aatatcaaat
361 taggaatcgt agtttcatga ttttctgtta cacctaactt tttgtgtggt gccctcctcc
421 ttgtcaatat taatgttaaa gtgcaattct ttttccttat cacgttgagc cattagtatc
481 aatttgctta cctgtattcc tttactatcc tcctttttct ccttcttgat aaatgtatgt
541 agattgcgta tatagtttcg tctaccctat gaacatattc cattttgtaa tttcgtgtcg
601 tttctattat gaatttcatt tataaagttt atgtacaaat atcataaaaa aagagaatct
661 ttttaagcaa ggattttctt aacttcttcg gcgacagcat caccgacttc ggtggtactg
721 ttggaaccac ctaaatcacc agttctgata cctgcatcca aaaccttttt aactgcatct
781 tcaatggcct taccttcttc aggcaagttc aatgacaatt tcaacatcat tgcagcagac
841 aagatagtgg cgatagggtc aaccttattc tttggcaaat ctggagcaga accgtggcat
901 ggttcgtaca aaccaaatgc ggtgttcttg tctggcaaag aggccaagga cgcagatggc
961 aacaaaccca aggaacctgg gataacggag gcttcatcgg agatgatatc accaaacatg
1021 ttgctggtga ttataatacc atttaggtgg gttgggttct taactaggat catggcggca
1081 gaatcaatca attgatgttg aaccttcaat gtagggaatt cgttcttgat ggtttcctcc
1141 acagtttttc tccataatct tgaagaggcc aaaacattag ctttatccaa ggaccaaata
1201 ggcaatggtg gctcatgttg tagggccatg aaagcggcca ttcttgtgat tctttgcact
1261 tctggaacgg tgtattgttc actatcccaa gcgacaccat caccatcgtc ttcctttctc
1321 ttaccaaagt aaatacctcc cactaattct ctgacaacaa cgaagtcagt acctttagca
1381 aattgtggct tgattggaga taagtctaaa agagagtcgg atgcaaagtt acatggtctt
1441 aagttggcgt acaattgaag ttctttacgg atttttagta aaccttgttc aggtctaaca
1501 ctaccggtac cccatttagg accacccaca gcacctaaca aaacggcatc aaccttcttg
1561 gaggcttcca gcgcctcatc tggaagtggg acacctgtag catcgatagc agcaccacca
1621 attaaatgat tttcgaaatc gaacttgaca ttggaacgaa catcagaaat agctttaaga
1681 accttaatgg cttcggctgt gatttcttga ccaacgtggt cacctggcaa aacgacgatc
1741 ttcttagggg cagacatagg ggcagacatt agaatggtat atccttgaaa tatatatata
1801 tattgctgaa atgtaaaagg taagaaaagt tagaaagtaa gacgattgct aaccacctat
1861 tggaaaaaac aataggtcct taaataatat tgtcaacttc aagtattgtg atgcaagcat
1921 ttagtcatga acgcttctct attctatatg aaaagccggt tccggcctct cacctttcct
1981 ttttctccca atttttcagt tgaaaaaggt atatgcgtca ggcgacctct gaaattaaca
2041 aaaaatttcc agtcatcgaa tttgattctg tgcgatagcg cccctgtgtg ttctcgttat
2101 gttgaggaaa aaaataatgg ttgctaagag attcgaactc ttgcatctta cgatacctga
2161 gtattcccac agttaactgc ggtcaagata tttcttgaat caggcgcctt agaccgctcg
2221 gccaaacaac caattacttg ttgagaaata gagtataatt atcctataaa tataacgttt
2281 ttgaacacac atgaacaagg aagtacagga caattgattt tgaagagaat gtggattttg
2341 atgtaattgt tgggattcca tttttaataa ggcaataata ttaggtatgt ggatatacta
2401 gaagttctcc tcgagggtcg atatgcggtg tgaaataccg cacagatgcg taaggagaaa
2461 ataccgcatc aggaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt
2521 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
2581 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
2641 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
2701 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
2761 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
2821 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
2881 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc gcgccattcg
2941 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
3001 cagctggcga aggggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
3061 cagtcacgac gttgtaaaac gacggccagt gaattgtaat acgactcact atagggcgaa
3121 ttggagctcc accgcggtgg cggccgctct agaactagtg gatcccccgg gctgcaggaa
3181 ttcgatatca agcttatcga taccgtcgac ctcgaggggg ggcccggtac ccagcttttg
3241 ttccctttag tgagggttaa ttccgagctt ggcgtaatca tggtcatagc tgtttcctgt
3301 gtgaaattgt tatccgctca caattccaca caacatagga gccggaagca taaagtgtaa
3361 agcctggggt gcctaatgag tgaggtaact cacattaatt gcgttgcgct cactgcccgc
3421 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
3481 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
3541 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
3601 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
3661 taaaaaggcc gcgttgctgg cgtttttcca taggctcggc ccccctgacg agcatcacaa
3721 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
3781 cccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
3841 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
3901 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
3961 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
4021 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
4081 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
4141 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
4201 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
4261 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
4321 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
4381 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
4441 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
4501 catagttgcc tgactgcccg tcgtgtagat aactacgata cgggagggct taccatctgg
4561 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
4621 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
4681 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
4741 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
4801 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgaaaaaa
4861 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
4921 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
4981 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
5041 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
5101 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
5161 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
5221 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
5281 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
5341 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
5401 ggttccgcgc acatttcccc gaaaagtgcc acctgggtcc ttttcatcac gtgctataaa
5461 aataattata atttaaattt tttaatataa atatataaat taaaaataga aagtaaaaaa
5521 agaaattaaa gaaaaaatag tttttgtttt ccgaagatgt aaaagactct agggggatcg
5581 ccaacaaata ctacctttta tcttgctctt cctgctctca ggtattaatg ccgaattgtt
5641 tcatcttgtc tgtgtagaag accacacacg aaaatcctgt gattttacat tttacttatc
5701 gttaatcgaa tgtatatcta tttaatctgc ttttcttgtc taataaatat atatgtaaag
5761 tacgcttttt gttgaaattt tttaaacctt tgtttatttt tttttcttca ttccgtaact
5821 cttctacctt ctttatttac tttctaaaat ccaaatacaa aacataaaaa taaataaaca
5881 cagagtaaat tcccaaatta ttccatcatt aaaagatacg aggcgcgtgt aagttacagg
5941 caagcgatcc gtcctaagaa accattatta tcatgacatt aacctataaa aataggcgta
6001 tcacgaggcc ctttcgtc
//