LOCUS Exported 6677 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Express gene of interest under CB promoter. ACCESSION . VERSION . KEYWORDS pCDH-CB SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6677) TITLE lenti vector JOURNAL Unpublished REFERENCE 2 (bases 1 to 6677) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6677 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 234..414 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 458..583 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1076..1309 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1493..1537 /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 1798..1915 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" intron 2567..2663 /label=SV40 intron /note="modified SV40 intron with splice donor and acceptor sites" misc_feature 2772..3360 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2825..2845) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3243..3254) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 3434..3667 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 3739..3860 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3776..3795) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3830..3849 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 3879..4014 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3941..3960 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(4052..4068) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4052..4068) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4065..4087) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4076..4092 /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(4100..4130) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4145..4166 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4283..4300) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4454..5042) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4534..4553) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5213..6073) /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 5836..5855 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6074..6178) /gene="bla" /label=AmpR promoter primer_bind 6246..6264 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(6302..6324) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 6424..6443 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 6637..6659 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 6651..6668 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 6652..6668 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" ORIGIN 1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca 61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta 121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga 181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc 241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg 421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct 481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt 541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag 601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt 661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt 721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg 781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag 841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag 901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg 961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag 1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag 1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc 1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga 1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc 1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg 1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata 1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa 1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga 1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa 1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat 1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt 1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg 1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggttaacttt 1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc 1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga 1921 ttaattaagg tacctctggt cgttacataa cttacggtaa atggcccgcc tggctgaccg 1981 cccaacgacc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 2041 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 2101 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 2161 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctact 2221 cgaggccacg ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta 2281 tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg gggggggggc 2341 gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg 2401 cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc 2461 ggcggcccta taaaaagcga agcgcgcggc gggcgggagc gggatcagcc accgcggtgg 2521 cggccctaga gtcgatcgag gaactgaaaa accagaaagt taactggtaa gtttagtctt 2581 tttgtctttt atttcaggtc cccgatccgg tggtggtgca aatcaaagaa ctgctcctca 2641 gtggatgttg cctttacttc taggcctgta cggaagtgtt acttctgctc taaaagctgc 2701 ggaattgtac ccgcggccga tccaccggtc gggatccgct agcgtttaaa cgcggccgcg 2761 aattcgtcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta 2821 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta 2881 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt 2941 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg 3001 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt 3061 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag 3121 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc 3181 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc 3241 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc 3301 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc 3361 ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa 3421 gaaaaggggg gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct 3481 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg 3541 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt 3601 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc 3661 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga 3721 atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat 3781 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc 3841 aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca 3901 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 3961 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact 4021 tttgcagaga cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 4081 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 4141 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 4201 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 4261 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 4321 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 4381 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 4441 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 4501 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 4561 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 4621 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 4681 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 4741 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 4801 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 4861 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc 4921 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 4981 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 5041 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 5101 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 5161 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 5221 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 5281 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 5341 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 5401 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 5461 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 5521 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 5581 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 5641 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 5701 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 5761 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 5821 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 5881 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 5941 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 6001 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 6061 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 6121 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 6181 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct 6241 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa 6301 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga 6361 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact 6421 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca 6481 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt 6541 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg 6601 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga 6661 cggccagtgc caagctg //