LOCUS CMV-hRad51 4422 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses the Rad51 protein in mammalian cells under CMV promoter.
ACCESSION .
VERSION .
KEYWORDS CMV-hRad51
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4422)
AUTHORS Rees HA, Yeh WH, Liu DR
TITLE Development of hRad51-Cas9 nickase fusions that mediate HDR without
double-stranded breaks.
JOURNAL Nat Commun. 2019 May 17;10(1):2212. doi: 10.1038/s41467-019-09983-4.
PUBMED 31101808
REFERENCE 2 (bases 1 to 4422)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
//www.snapgene.com
FEATURES Location/Qualifiers
source 1..4422
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 132..335
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 286..306
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 377..396
/label=T7
/note="T7 promoter, forward primer"
promoter 377..395
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 403..412
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
primer_bind complement(1477..1494)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1483..1707
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(1778..1794)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1778..1794)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1791..1813)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1802..1818
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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(1826..1856)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1871..1892
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2009..2026)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2180..2768)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2260..2279)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2939..3799)
/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 3562..3581
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3800..3904)
/gene="bla"
/label=AmpR promoter
primer_bind complement(3983..4002)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 4174..131
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
ORIGIN
1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
361 agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat ggcgatgcag
421 atgcagttgg aagcgaatgc agatactagt gtcgaggaag agtcatttgg cccgcaaccc
481 atctcgcgtt tagagcaatg tggcatcaat gcaaacgatg tgaaaaaatt agaggaagct
541 ggattccaca cggtcgaagc ggtcgcatac gcaccgaaaa aagagctgat caacatcaaa
601 ggcatcagcg aggcgaaagc cgataagatt cttgcagagg cggcgaaatt agttcccatg
661 ggatttacga cggcgactga gttccatcaa cgtcgttccg agatcattca aatcacgacc
721 ggaagcaagg agttggataa actgcttcaa ggaggcatcg aaactggctc gatcactgaa
781 atgtttggag agtttcgtac tggcaaaacc cagatctgcc acacattggc cgtcacttgc
841 cagcttccga tcgatcgtgg tgggggcgag gggaaggcca tgtacattga tactgagggg
901 acgtttcgtc ccgaacgcct tcttgcggtg gcagagcgtt acggcttgtc agggtctgat
961 gttcttgaca atgttgccta tgctcgcgca tttaataccg accaccagac gcagcttctg
1021 taccaagcaa gcgcgatgat ggtagagtct cgttacgctt tattgattgt tgactcagca
1081 acagcgttgt accgcacaga ttattctggg cgcggcgaat tgtcggcacg ccaaatgcat
1141 ttagcccgct ttttgcgtat gttgctgcgc cttgcagacg aatttggggt ggctgtggtg
1201 atcaccaacc aagttgtagc acaggtggac ggtgccgcta tgtttgccgc tgatccgaaa
1261 aaacccatcg gagggaatat tatcgcacat gcatcgacaa ctcgcttata tctgcgtaag
1321 ggccgtggag aaacacgcat ctgcaaaatc tatgactccc catgcctgcc ggaggcggaa
1381 gccatgttcg caattaacgc agatggcgta ggggacgcca aagattgatg actggtacac
1441 atcaccatca ccattgagtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
1501 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
1561 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
1621 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
1681 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcga
1741 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
1801 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
1861 agggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
1921 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
1981 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
2041 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
2101 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
2161 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
2221 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
2281 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
2341 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
2401 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
2461 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
2521 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
2581 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
2641 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
2701 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
2761 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
2821 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
2881 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
2941 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
3001 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
3061 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
3121 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
3181 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
3241 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
3301 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
3361 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
3421 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
3481 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
3541 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
3601 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
3661 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
3721 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
3781 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
3841 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
3901 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat cgatctcccg
3961 atcccctagg gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
4021 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
4081 caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc
4141 tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact agttattaat
4201 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
4261 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
4321 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
4381 atttacggta aactgcccac ttggcagtac atcaagtgta tc
//
