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LOCUS pLVX-IRES-Puro-3xHA-C 8228 bp DNA circular SYN 13-JUL-2023
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8228)
AUTHORS Biofeng
TITLE Direct Submission
JOURNAL Exported Jul 13, 2023 from SnapGene 6.0.2
https://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8228
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 1..634
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 681..806
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1303..1536
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1721..1765
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 2028..2143
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)"
enhancer 2201..2504
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2505..2708
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 2836..2916
/codon_start=1
/product="three tandem HA epitope tags"
/label=3xHA
/translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
misc_feature 2931..3504
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 3537..4136
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
misc_feature 4150..4738
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS complement(4621..4632)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 4945..5578
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
primer_bind complement(5706..5722)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 5730..5746
/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(5754..5784)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5799..5820
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(6108..6696)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6867..7727)
/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"
promoter complement(7728..7832)
/gene="bla"
/label=AmpR promoter
polyA_signal 7880..8014
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
ORIGIN
1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
2761 tccatagaag acaccgactc tactagagga tctatttccg gtgaattcct cgagactagt
2821 tctagagcgg ccgcgtaccc atatgacgtt ccagactacg cgtatccgta cgacgttccg
2881 gattacgctt acccttacga tgtacctgac tacgcttaat agggatcccg cccctctccc
2941 tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc
3001 tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc
3061 cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt
3121 ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct
3181 gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa
3241 aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt
3301 tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag
3361 gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt
3421 acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt
3481 ttcctttgaa aaacacgatg ataagcttgc cacaacccac aaggagacga ccttccatga
3541 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca
3601 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc
3661 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
3721 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
3781 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
3841 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
3901 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
3961 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
4021 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
4081 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctagacgc
4141 gtctggaaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
4201 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
4261 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
4321 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
4381 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
4441 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
4501 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
4561 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
4621 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
4681 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
4741 ggaattaatt ctgcagtcga gacctagaaa aacatggagc aatcacaagt agcaatacag
4801 cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc
4861 cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
4921 actttttaaa agaaaagagg ggactggaag ggctaattca ctcccaacga agacaagata
4981 tccttgatct gtggatctac cacacacaag gctacttccc tgattagcag aactacacac
5041 cagggccagg ggtcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg
5101 agccagataa ggtagaagag gccaataaag gagagaacac cagcttgtta caccctgtga
5161 gcctgcatgg gatggatgac ccggagagag aagtgttaga gtggaggttt gacagccgcc
5221 tagcatttca tcacgtggcc cgagagctgc atccggagta cttcaagaac tgctgatatc
5281 gagcttgcta caagggactt tccgctgggg actttccagg gaggcgtggc ctgggcggga
5341 ctggggagtg gcgagccctc agatcctgca tataagcagc tgctttttgc ctgtactggg
5401 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
5461 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
5521 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
5581 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag
5641 agtgagaggc cttgacattg ctagcgttta ccgtcgacct ctagctagag cttggcgtaa
5701 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
5761 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
5821 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
5881 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
5941 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
6001 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
6061 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
6121 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
6181 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
6241 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
6301 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
6361 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
6421 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
6481 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
6541 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
6601 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
6661 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
6721 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
6781 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
6841 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
6901 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
6961 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
7021 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
7081 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
7141 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
7201 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
7261 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
7321 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
7381 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
7441 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
7501 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
7561 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
7621 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
7681 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
7741 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
7801 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
7861 gtcgacggat cgggagatca acttgtttat tgcagcttat aatggttaca aataaagcaa
7921 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
7981 caaactcatc aatgtatctt atcatgtctg gatcaactgg ataactcaag ctaaccaaaa
8041 tcatcccaaa cttcccaccc cataccctat taccactgcc aattacctgt ggtttcattt
8101 actctaaacc tgtgattcct ctgaattatt ttcattttaa agaaattgta tttgttaaat
8161 atgtactaca aacttagtag tttttaaaga aattgtattt gttaaatatg tactacaaac
8221 ttagtagt
//
