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pFA6a-kanMX6.gb
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LOCUS pFA6a-kanMX6 3938 bp ds-DNA circular SYN 16-JUN-2022
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS pFA6a-kanMX6.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 3938)
AUTHORS Bahler J, Wu JQ, Longtine MS, Shah NG, McKenzie A 3rd, Steever AB,
Wach A, Philippsen P, Pringle JR
TITLE Heterologous modules for efficient and versatile PCR-based gene
targeting in Schizosaccharomyces pombe.
JOURNAL Yeast. 1998 Jul;14(10):943-51.
PUBMED 9717240
REFERENCE 2 (bases 1 to 3938)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jun 16, 2022 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3938
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(35..52)
/label="L4440"
/note="L4440 vector, forward primer"
rep_origin complement(206..794)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(286..305)
/label="pBR322ori-F"
/note="pBR322 origin, forward primer"
CDS complement(965..1825)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 1588..1607
/label="Amp-R"
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1826..1930)
/gene="bla"
/label="AmpR promoter"
primer_bind 1998..2016
/label="pBRforEco"
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(2054..2076)
/label="pGEX 3'"
/note="pGEX vectors, reverse primer"
primer_bind 2176..2195
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 2276..2294
/label="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 2276..2293
/label="SP6"
/note="SP6 promoter, forward primer"
gene 2407..3763
/label="kanMX"
/note="yeast selectable marker conferring kanamycin
resistance (Wach et al., 1994)"
promoter 2407..2750
/label="TEF promoter"
/note="Ashbya gossypii TEF promoter"
CDS 2751..3560
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label="KanR"
/note="confers resistance to kanamycin"
/translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind complement(2818..2837)
/label="Kan-R"
/note="Kanamycin resistance gene, reverse primer"
terminator 3566..3763
/label="TEF terminator"
/note="Ashbya gossypii TEF terminator"
primer_bind complement(3867..3886)
/label="T7"
/note="T7 promoter, forward primer"
promoter complement(3868..3886)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
61 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
121 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
181 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
241 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
301 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
361 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
421 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
481 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
541 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
601 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
661 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
721 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
781 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
841 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
901 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
961 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
1021 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
1081 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
1141 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
1201 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
1261 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
1321 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
1381 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
1441 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
1501 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
1561 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
1621 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
1681 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
1741 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
1801 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
1861 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
1921 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
1981 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
2041 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
2101 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
2161 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atggacatat
2221 tgtcgttaga acgcggctac aattaataca taaccttatg tatcatacac atacgattta
2281 ggtgacacta tagaacgcgg ccgccagctg aagcttcgta cgctgcaggt cgacggatcc
2341 ccgggttaat taaggcgcgc cagatctgtt tagcttgcct cgtccccgcc gggtcacccg
2401 gccagcgaca tggaggccca gaataccctc cttgacagtc ttgacgtgcg cagctcaggg
2461 gcatgatgtg actgtcgccc gtacatttag cccatacatc cccatgtata atcatttgca
2521 tccatacatt ttgatggccg cacggcgcga agcaaaaatt acggctcctc gctgcagacc
2581 tgcgagcagg gaaacgctcc cctcacagac gcgttgaatt gtccccacgc cgcgcccctg
2641 tagagaaata taaaaggtta ggatttgcca ctgaggttct tctttcatat acttcctttt
2701 aaaatcttgc taggatacag ttctcacatc acatccgaac ataaacaacc atgggtaagg
2761 aaaagactca cgtttcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt
2821 ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga
2881 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta
2941 cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc
3001 attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc ggcaaaacag
3061 cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag
3121 tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg
3181 tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt
3241 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataagcttt
3301 tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt
3361 ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat
3421 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
3481 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga
3541 tgctcgatga gtttttctaa tcagtactga caataaaaag attcttgttt tcaagaactt
3601 gtcatttgta tagttttttt atattgtagt tgttctattt taatcaaatg ttagcgtgat
3661 ttatattttt tttcgcctcg acatcatctg cccagatgcg aagttaagtg cgcagaaagt
3721 aatatcatgc gtcaatcgta tgtgaatgct ggtcgctata ctgctgtcga ttcgatacta
3781 acgccgccat ccagtttaaa cgagctcgaa ttcatcgatg atatcagatc cactagtggc
3841 ctatgcggcc gcggatctgc cggtctccct atagtgagtc gtattaattt cgataagcca
3901 ggttaacctg cattaatgaa tcggccaacg cgcgggga
//