LOCUS Exported 6427 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Express gene of interest under truncated EF-1 promoter. ACCESSION . VERSION . KEYWORDS pCDH-EF1s SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6427) TITLE express gene of interest under truncated EF-1 promoter JOURNAL Unpublished REFERENCE 2 (bases 1 to 6427) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6427 /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 1804..1920 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 1961..2172 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" LTR 2185..2453 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" misc_feature 2522..3110 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2575..2595) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2993..3004) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 3184..3417 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 3489..3610 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3526..3545) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3580..3599 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 3629..3764 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3691..3710 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(3802..3818) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3802..3818) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3815..3837) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3826..3842 /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(3850..3880) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3895..3916 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4033..4050) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4204..4792) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4284..4303) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4963..5823) /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 5586..5605 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5824..5928) /gene="bla" /label=AmpR promoter primer_bind 5996..6014 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(6052..6074) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 6174..6193 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 6387..6409 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 6401..6418 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 6402..6418 /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 ggtatcggtt 1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat 1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt 1921 atcgatacaa ggatctgcga tcgctccggt gcccgtcagt gggcagagcg cacatcgccc 1981 acagtccccg agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg 2041 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg 2101 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc 2161 gccagaacac agctgaagct tcgaggggct cgcatctctc cttcacgcgc ccgccgccct 2221 acctgaggcc gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc 2281 tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt 2341 ccggcgctcc cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg 2401 cttgctcaac tctacgtctt tgtttcgttt tctgttctgc gccgttacag atccaagctg 2461 tgaccggcgc ctacgctaga cggatccgct agcgtttaaa cgcggccgcg aattcgtcga 2521 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 2581 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 2641 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 2701 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 2761 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 2821 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 2881 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct 2941 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 3001 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 3061 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt 3121 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg 3181 gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt 3241 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc 3301 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg 3361 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta 3421 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga 3481 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 3541 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 3601 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca 3661 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc 3721 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga 3781 cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 3841 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 3901 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 3961 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 4021 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 4081 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 4141 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 4201 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 4261 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 4321 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 4381 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 4441 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 4501 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 4561 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 4621 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 4681 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 4741 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 4801 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 4861 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 4921 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 4981 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 5041 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 5101 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 5161 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 5221 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 5281 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 5341 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 5401 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 5461 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 5521 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 5581 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 5641 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 5701 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 5761 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 5821 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 5881 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 5941 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 6001 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 6061 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 6121 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 6181 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 6241 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 6301 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 6361 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 6421 caagctg //