LOCUS Exported 6630 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Lentiviral system for reduced ectopic expression.. ACCESSION . VERSION . KEYWORDS pLVX-CMV200 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6630) AUTHORS Dean KM, Roudot P, Reis CR, Welf ES, Mettlen M, Fiolka R TITLE Diagonally Scanned Light-Sheet Microscopy for Fast Volumetric Imaging of Adherent Cells. JOURNAL Biophys J. 2016 Mar 29;110(6):1456-65. doi: 10.1016/j.bpj.2016.01.029. PUBMED 27028654 REFERENCE 2 (bases 1 to 6630) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6630 /organism="synthetic DNA construct" /mol_type="other DNA" 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)" misc_feature 2402..2458 /label=MCS /note="multiple cloning site" misc_feature 2599..3187 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2652..2672) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3070..3081) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 3394..4027 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" primer_bind complement(4155..4171) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4155..4171) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4168..4190) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4179..4195 /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(4203..4233) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4248..4269 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4386..4403) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4557..5145) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4637..4656) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5316..6176) /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 5939..5958 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6177..6281) /gene="bla" /label=AmpR promoter polyA_signal 6329..6463 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6366..6385) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6420..6439 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" 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 cgatagttat taatagtaat caattacggg gtcattagtt 2221 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga 2281 ccgcccaacg acccccgccc attgacgtca ataatgacgt acgttcccat agtaacgcca 2341 atagggactt tccattgacg tcaatgggtg gagtatttac ggtccgctag cgctaactag 2401 tagatctcga gctcaagctt cgaattctgc agtcgacggt accgcgggcc cgggatccac 2461 cggatctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt 2521 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt 2581 taacgacgcg tctggaacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt 2641 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat 2701 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg 2761 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt 2821 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact 2881 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc 2941 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg 3001 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc 3061 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg 3121 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc 3181 tccccgcctg gaattaattc tgcagtcgag acctagaaaa acatggagca atcacaagta 3241 gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg 3301 tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag 3361 atcttagcca ctttttaaaa gaaaagaggg gactggaagg gctaattcac tcccaacgaa 3421 gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga 3481 actacacacc agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag 3541 taccagttga gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac 3601 accctgtgag cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg 3661 acagccgcct agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact 3721 gctgatatcg agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc 3781 tgggcgggac tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc 3841 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg 3901 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt 3961 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc 4021 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga 4081 atatcagaga gtgagaggcc ttgacattgc tagcgtttac cgtcgacctc tagctagagc 4141 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca 4201 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa 4261 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag 4321 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc 4381 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct 4441 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg 4501 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 4561 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga 4621 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 4681 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 4741 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 4801 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 4861 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac 4921 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 4981 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc 5041 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 5101 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 5161 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 5221 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 5281 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca 5341 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag 5401 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac 5461 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc 5521 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct 5581 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc 5641 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg 5701 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc 5761 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat 5821 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag 5881 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat 5941 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg 6001 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca 6061 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga 6121 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc 6181 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata 6241 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg 6301 ccacctgacg tcgacggatc gggagatcaa cttgtttatt gcagcttata atggttacaa 6361 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 6421 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc 6481 taaccaaaat catcccaaac ttcccacccc ataccctatt accactgcca attacctgtg 6541 gtttcattta ctctaaacct gtgattcctc tgaattattt tcattttaaa gaaattgtat 6601 ttgttaaata tgtactacaa acttagtagt //