LOCUS Exported 8858 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION For LIC cloning with N-terminal V5 tag. ACCESSION . VERSION . KEYWORDS pLV-EF1a-V5-LIC SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8858) TITLE Gerace lab plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 8858) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8858 /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)" promoter 2221..2432 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" intron 2433..3371 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 3328..3348 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" regulatory 3387..3396 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3396..3437 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" primer_bind complement(3749..3766) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 3976..3995 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 4157..4756 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(4157..4176) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 4653..4673 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 4780..5368 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4833..4853) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5251..5262) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 5576..6209 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" primer_bind complement(6338..6354) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6338..6354) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6351..6373) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6362..6378 /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(6386..6416) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6431..6452 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6569..6586) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6740..7325) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6820..6839) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7496..8356) /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 8119..8138 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8357..8461) /gene="bla" /label=AmpR promoter polyA_signal 8509..8643 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(8546..8565) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 8600..8619 /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 actacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg 2161 acagcagaga tccagtttat cgatgaggcc ctttcgtctt cactcgaggt gcccgtcagt 2221 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa 2281 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc 2341 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc 2401 tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc 2461 ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc tggctgcagt 2521 acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg 2581 cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc 2641 gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca 2701 tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg 2761 cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc 2821 gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg 2881 gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta 2941 tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat 3001 ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc tcgggagagc 3061 gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat 3121 gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg agcttttgga 3181 gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt 3241 gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc 3301 tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc 3361 ttccatttca ggtgtcgtga ggatccgccg ccatgggtaa gcctatccct aaccctctcc 3421 tcggtctcga ttctacgggg ggcggagggg gtgggcccgg gccgcgacga tgtttaaact 3481 tcgaaacgcg tgaattcctc gagggcggcc gctctagagt cgacgggccg cggtaacaat 3541 tgttaactaa cttaagctag caacggtttc cctctagcgg gatcaattcc gccccccccc 3601 cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta 3661 ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc 3721 ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat 3781 gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc 3841 ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt 3901 gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt 3961 gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag 4021 aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt 4081 tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga 4141 aaaacacgat aataccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga 4201 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca 4261 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac 4321 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt 4381 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat 4441 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc 4501 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca 4561 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg 4621 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg 4681 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg 4741 caagcccggt gcctgacccg ggcggggcgc gtctggaaca atcaacctct ggattacaaa 4801 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac 4861 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc 4921 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt 4981 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc 5041 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc 5101 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg 5161 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt 5221 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc 5281 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt 5341 cggatctccc tttgggccgc ctccccgcct ggaattaatt ctgcagtcga gacctagaaa 5401 aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag 5461 aagcacaaga ggaggaggag gtgggttttt ccagtcacac ctcaggtacc tttaagacca 5521 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaagag gggactggaa 5581 gggctaattc actcccaacg aagacaagat atccttgatc tgtggatcta ccacacacaa 5641 ggctacttcc ctgattagca gaactacaca ccagggccag gggtcagata tccactgacc 5701 tttggatggt gctacaagct agtaccagtt gagccagata aggtagaaga ggccaataaa 5761 ggagagaaca ccagcttgtt acaccctgtg agcctgcatg ggatggatga cccggagaga 5821 gaagtgttag agtggaggtt tgacagccgc ctagcatttc atcacgtggc ccgagagctg 5881 catccggagt acttcaagaa ctgctgatat cgagcttgct acaagggact ttccgctggg 5941 gactttccag ggaggcgtgg cctgggcggg actggggagt ggcgagccct cagatcctgc 6001 atataagcag ctgctttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg 6061 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt 6121 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc 6181 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta 6241 tttataactt gcaaagaaat gaatatcaga gagtgagagg ccttgacatt gctagcgttt 6301 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa 6361 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct 6421 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc 6481 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg 6541 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc 6601 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag 6661 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa 6721 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc 6781 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc 6841 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg 6901 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt 6961 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc 7021 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc 7081 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag 7141 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg 7201 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa 7261 ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat 7321 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac 7381 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt 7441 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc 7501 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg 7561 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg 7621 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc 7681 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta 7741 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg 7801 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct 7861 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta 7921 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg 7981 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga 8041 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt 8101 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca 8161 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt 8221 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt 8281 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga 8341 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt 8401 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc 8461 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatcaa cttgtttatt 8521 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt 8581 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg 8641 atcaactgga taactcaagc taaccaaaat catcccaaac ttcccacccc ataccctatt 8701 accactgcca attacctagt ggtttcattt actctaaacc tgtgattcct ctgaattatt 8761 ttcattttaa agaaattgta tttgttaaat atgtactaca aacttagtag tttttaaaga 8821 aattgtattt gttaaatatg tactacaaac ttagtagt //