LOCUS Exported 5797 bp ds-DNA circular SYN 25-MAY-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pRS423
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5797)
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..5797
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 68..1410
/note="2u ori"
/note="yeast 2u plasmid origin of replication"
protein_bind 400..447
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FRT"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
promoter 1437..1541
/gene="bla"
/note="AmpR promoter"
CDS 1542..2402
/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"
rep_origin 2573..3161
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 3485..3515
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 3523..3539
/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)."
primer_bind 3547..3563
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
CDS 3559..4137
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/note="lacZ-alpha"
/translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
TDRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFA
GFPRQALNRGLPLGFRFSALRHLDPKKLD"
promoter 3584..3602
/note="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
misc_feature 3615..3722
/note="MCS"
/note="pBluescript multiple cloning site"
primer_bind 3639..3655
/note="SK primer"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3689..3705)
/note="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(3731..3749)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3759..3775)
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 3920..4375
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(4635..5294)
/codon_start=1
/gene="S. cerevisiae HIS3"
/product="imidazoleglycerol-phosphate dehydratase, required
for histidine biosynthesis"
/note="HIS3"
/note="yeast auxotrophic marker"
/translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
promoter complement(5295..5481)
/gene="S. cerevisiae HIS3"
/note="HIS3 promoter"
ORIGIN
1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
61 cttagatgat ccaatatcaa aggaaatgat agcattgaag gatgagacta atccaattga
121 ggagtggcag catatagaac agctaaaggg tagtgctgaa ggaagcatac gataccccgc
181 atggaatggg ataatatcac aggaggtact agactacctt tcatcctaca taaatagacg
241 catataagta cgcatttaag cataaacacg cactatgccg ttcttctcat gtatatatat
301 atacaggcaa cacgcagata taggtgcgac gtgaacagtg agctgtatgt gcgcagctcg
361 cgttgcattt tcggaagcgc tcgttttcgg aaacgctttg aagttcctat tccgaagttc
421 ctattctcta gaaagtatag gaacttcaga gcgcttttga aaaccaaaag cgctctgaag
481 acgcactttc aaaaaaccaa aaacgcaccg gactgtaacg agctactaaa atattgcgaa
541 taccgcttcc acaaacattg ctcaaaagta tctctttgct atatatctct gtgctatatc
601 cctatataac ctacccatcc acctttcgct ccttgaactt gcatctaaac tcgacctcta
661 cattttttat gtttatctct agtattactc tttagacaaa aaaattgtag taagaactat
721 tcatagagtg aatcgaaaac aatacgaaaa tgtaaacatt tcctatacgt agtatataga
781 gacaaaatag aagaaaccgt tcataatttt ctgaccaatg aagaatcatc aacgctatca
841 ctttctgttc acaaagtatg cgcaatccac atcggtatag aatataatcg gggatgcctt
901 tatcttgaaa aaatgcaccc gcagcttcgc tagtaatcag taaacgcggg aagtggagtc
961 aggctttttt tatggaagag aaaatagaca ccaaagtagc cttcttctaa ccttaacgga
1021 cctacagtgc aaaaagttat caagagactg cattatagag cgcacaaagg agaaaaaaag
1081 taatctaaga tgctttgtta gaaaaatagc gctctcggga tgcatttttg tagaacaaaa
1141 aagaagtata gattctttgt tggtaaaata gcgctctcgc gttgcatttc tgttctgtaa
1201 aaatgcagct cagattcttt gtttgaaaaa ttagcgctct cgcgttgcat ttttgtttta
1261 caaaaatgaa gcacagattc ttcgttggta aaatagcgct ttcgcgttgc atttctgttc
1321 tgtaaaaatg cagctcagat tctttgtttg aaaaattagc gctctcgcgt tgcatttttg
1381 ttctacaaaa tgaagcacag atgcttcgtt caggtggcac ttttcgggga aatgtgcgcg
1441 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
1501 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
1561 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
1621 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
1681 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
1741 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
1801 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
1861 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
1921 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
1981 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
2041 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
2101 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
2161 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
2221 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
2281 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
2341 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
2401 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
2461 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
2521 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
2581 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
2641 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
2701 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
2761 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
2821 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
2881 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
2941 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
3001 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
3061 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
3121 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
3181 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
3241 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
3301 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
3361 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
3421 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta cctcactcat taggcacccc
3481 aggctttaca ctttatgctt ccggctccta tgttgtgtgg aattgtgagc ggataacaat
3541 ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa
3601 gggaacaaaa gctggagctc caccgcggtg gcggccgctc tagaactagt ggatcccccg
3661 ggctgcagga attcgatatc aagcttatcg ataccgtcga cctcgagggg gggcccggta
3721 cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt
3781 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
3841 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
3901 ctgaatggcg aatggcgcga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
3961 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
4021 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
4081 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
4141 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
4201 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
4261 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
4321 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aatttcctga
4381 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatagat ccgtcgagtt
4441 caagagaaaa aaaaagaaaa agcaaaaaga aaaaaggaaa gcgcgcctcg ttcagaatga
4501 cacgtataga atgatgcatt accttgtcat cttcagtatc atactgttcg tatacatact
4561 tactgacatt cataggtata catatataca catgtatata tatcgtatgc tgcagcttta
4621 aataatcggt gtcactacat aagaacacct ttggtggagg gaacatcgtt ggtaccattg
4681 ggcgaggtgg cttctcttat ggcaaccgca agagccttga acgcactctc actacggtga
4741 tgatcattct tgcctcgcag acaatcaacg tggagggtaa ttctgctagc ctctgcaaag
4801 ctttcaagaa aatgcgggat catctcgcaa gagagatctc ctactttctc cctttgcaaa
4861 ccaagttcga caactgcgta cggcctgttc gaaagatcta ccaccgctct ggaaagtgcc
4921 tcatccaaag gcgcaaatcc tgatccaaac ctttttactc cacgcgccag tagggcctct
4981 ttaaaagctt gaccgagagc aatcccgcag tcttcagtgg tgtgatggtc gtctatgtgt
5041 aagtcaccaa tgcactcaac gattagcgac cagccggaat gcttggccag agcatgtatc
5101 atatggtcca gaaaccctat acctgtgtgg acgttaatca cttgcgattg tgtggcctgt
5161 tctgctactg cttctgcctc tttttctggg aagatcgagt gctctatcgc taggggacca
5221 ccctttaaag agatcgcaat ctgaatcttg gtttcatttg taatacgctt tactagggct
5281 ttctgctctg tcatctttgc cttcgtttat cttgcctgct cattttttag tatattcttc
5341 gaagaaatca cattacttta tataatgtat aattcattat gtgataatgc caatcgctaa
5401 gaaaaaaaaa gagtcatccg ctaggggaaa aaaaaaaatg aaaatcatta ccgaggcata
5461 aaaaaatata gagtgtacta gaggaggcca agagtaatag aaaaagaaaa ttgcgggaaa
5521 ggactgtgtt atgacttccc tgactaatgc cgtgttcaaa cgatacctgg cagtgactcc
5581 tagcgctcac caagctctta aaacgggaat ttatggtgca ctctcagtac aatctgctct
5641 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
5701 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
5761 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcga
//