LOCUS Ub-G76V-YFP 4959 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Ub-G76V-YFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4959)
AUTHORS Menendez-Benito V, Verhoef LG, Masucci MG, Dantuma NP
TITLE Endoplasmic reticulum stress compromises the ubiquitin-proteasome
system.
JOURNAL Hum Mol Genet. 2005 Oct 1. 14(19):2787-99.
PUBMED 16103128
REFERENCE 2 (bases 1 to 4959)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4959
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 61..364
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 365..568
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 519..539
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
CDS 638..862
/codon_start=1
/product="human ubiquitin"
/label=ubiquitin
/translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
AGKQLEDGRTLSDYNIQKESTLHLVLRLRG"
regulatory 899..908
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 905..1624
/codon_start=1
/product="enhanced YFP"
/label=EYFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(950..971)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1211..1230)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1558..1579
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 1747..1868
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1784..1803)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1838..1857
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(1875..2330)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2012..2033)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2224..2243
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2357..2461
/gene="bla"
/label=AmpR promoter
promoter 2463..2820
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2486..2506)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2671..2806
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2733..2752
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2855..3649
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(2909..2928)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3519..3538
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3837..3856)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3881..3928
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4257..4845
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4746..4765
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
601 ccggactcag atctcgagct caagcttcga attcaccatg cagatcttcg tgaagactct
661 gactggtaag accatcaccc tcgaggttga gcccagtgac accattgaga atgtcaaggc
721 aaagatccaa gataaggaag gcatccctcc tgaccagcag aggctgatct ttgctggaaa
781 acagctggaa gatgggcgca ccctgtctga ctacaacatc cagaaagagt ccaccctgca
841 cctggtactc cgtctcagag gtgtggtggg gaagcttggt cgacaggatc caccggtcgc
901 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
961 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
1021 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
1081 caccctcgtg accaccttcg gctacggcct gaagtgcttc gcccgctacc ccgaccacat
1141 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
1201 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
1261 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
1321 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
1381 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
1441 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
1501 ccactacctg agctaccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
1561 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
1621 gtaaagcggc cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt
1681 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
1741 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
1801 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
1861 gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
1921 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
1981 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
2041 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
2101 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
2161 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
2221 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
2281 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
2341 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
2401 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
2461 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
2521 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
2581 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
2641 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
2701 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
2761 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
2821 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
2881 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
2941 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
3001 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
3061 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
3121 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
3181 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
3241 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
3301 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
3361 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
3421 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
3481 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
3541 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
3601 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
3661 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
3721 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
3781 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
3841 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
3901 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
3961 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
4021 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
4081 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
4141 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
4201 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
4261 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
4321 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
4381 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
4441 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
4501 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
4561 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
4621 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
4681 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
4741 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
4801 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
4861 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
4921 tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
