LOCUS Exported 7139 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Express gene of interest under EF-1 promoter. ACCESSION . VERSION . KEYWORDS pCDH-EF1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7139) TITLE lenti vector JOURNAL Unpublished REFERENCE 2 (bases 1 to 7139) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7139 /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..1921 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2019..3197 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 2250..3188 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 3145..3165 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" misc_feature 3234..3822 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3287..3307) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3705..3716) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 3896..4129 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 4201..4322 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4238..4257) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4292..4311 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 4341..4476 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4403..4422 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(4514..4530) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4514..4530) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4527..4549) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4538..4554 /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(4562..4592) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4607..4628 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4745..4762) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4916..5504) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4996..5015) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5675..6535) /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 6298..6317 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6536..6640) /gene="bla" /label=AmpR promoter primer_bind 6708..6726 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(6764..6786) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 6886..6905 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 7099..7121 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 7113..7130 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 7114..7130 /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 attacaaaaa ttcaaaattt 1921 tatcgatatc gatagctttg caaagatgga taaagtttta aacagagagg aatctttgca 1981 gctaatggac cttctaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg 2041 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaaccg 2101 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc 2161 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt 2221 ttcgcaacgg gtttgccgcc agaacacagg taagtgccgt gtgtggttcc cgcgggcctg 2281 gcctctttac gggttatggc ccttgcgtgc cttgaattac ttccacctgg ctgcagtacg 2341 tgattcttga tcccgagctt cgggttggaa gtgggtggga gagttcgagg ccttgcgctt 2401 aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg 2461 tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt 2521 aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg 2581 gccaagatct gcacactggt atttcggttt ttggggccgc gggcggcgac ggggcccgtg 2641 cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac 2701 gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg 2761 ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc 2821 cgcttcccgg ccctgctgca gggagctcaa aatggaggac gcggcgctcg ggagagcggg 2881 cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg 2941 actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctcgagc ttttggagta 3001 cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg 3061 tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa tttgcccttt 3121 ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc 3181 catttcaggt gtcgtgagga attggatccc cgggcggccg cgaattcgtc gacaatcaac 3241 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta 3301 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt 3361 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg 3421 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg 3481 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca 3541 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca 3601 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg 3661 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag 3721 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc 3781 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggtacc tttaagacca 3841 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa 3901 gggctaattc actcccaacg aaaataagat ctgctttttg cttgtactgg gtctctctgg 3961 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct 4021 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt 4081 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca 4141 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg 4201 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca 4261 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct 4321 tatcatgtct ggctctagct atcccgcccc taactccgcc cagttccgcc cattctccgc 4381 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc 4441 tattccagaa gtagtgagga ggcttttttg gaggcctaga cttttgcaga gacggcccaa 4501 attcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac 4561 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac 4621 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc 4681 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg 4741 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc 4801 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt 4861 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 4921 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa 4981 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc 5041 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg 5101 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc 5161 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc 5221 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca 5281 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 5341 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 5401 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt 5461 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 5521 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 5581 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa 5641 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac 5701 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 5761 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc 5821 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca 5881 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta 5941 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg 6001 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc 6061 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg 6121 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt 6181 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt 6241 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata 6301 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc 6361 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac 6421 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa 6481 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct 6541 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat 6601 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc 6661 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca 6721 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc 6781 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg 6841 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga 6901 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat 6961 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc 7021 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt 7081 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gccaagctg //