LOCUS Exported 7647 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Lentiviral empty cloning vector containing IRES and GFP. ACCESSION . VERSION . KEYWORDS pCIG3 (pCMV-IRES-GFP version 3) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7647) AUTHORS Caviness K, Cicchini L, Rak M, Umashankar M, Goodrum F TITLE Complex expression of the UL136 gene of human cytomegalovirus results in multiple protein isoforms with unique roles in replication. JOURNAL J Virol. 2014 Dec;88(24):14412-25. doi: 10.1128/JVI.02711-14. Epub 2014 Oct 8. PUBMED 25297993 REFERENCE 2 (bases 1 to 7647) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7647 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 6..232 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 233..413 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 460..585 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1078..1311 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1496..1540 /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 1807..1923 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2013..2216 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2167..2187 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 2213..2237 /label=LNCX /note="Human CMV promoter, forward primer" misc_feature 2388..2960 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(2554..2571) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 2781..2800 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" regulatory 2981..2990 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2987..3706 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 2987..3703 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3032..3053) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3293..3312) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3640..3661 /label=EGFP-C /note="EGFP, forward primer" misc_feature 3726..4314 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3779..3799) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4197..4208) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 4388..4621 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 4693..4814 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4730..4749) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4784..4803 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 4854..4989 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4916..4935 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(5022..5038) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5022..5038) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5035..5057) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5046..5062 /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(5070..5100) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5115..5136 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5253..5270) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5424..6012) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5504..5523) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6183..7043) /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 6806..6825 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7044..7148) /gene="bla" /label=AmpR promoter primer_bind 7216..7234 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(7272..7294) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 7394..7413 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 7607..7629 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 7621..7638 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 7622..7638 /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 cgatacataa acgggtctct 241 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa 301 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc 361 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc 421 ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg 481 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt 541 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg 601 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa 661 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg 721 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca 781 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa 841 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa 901 agtaagacca ccgcacagca agcggccact gatcttcaga cctggaggag gagatatgag 961 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt 1021 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg 1081 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac 1141 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct 1201 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct 1261 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg 1321 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa 1381 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa 1441 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa 1501 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac 1561 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag 1621 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc 1681 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga 1741 aggtggagag agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg 1801 gttaactttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga 1861 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aattcaaaat 1921 tttatcgata ctagtattat gcccagtaca tgaccttatg ggactttcct acttggcagt 1981 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg 2041 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg 2101 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc 2161 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ttatataagc agagctcgtt 2221 tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagat 2281 tctagagcta gcgaattcgg tgatcagggt atacggctcg aggggatatc gggtttaaac 2341 ggggcgcgcc gggcgatcgc ggttcgaagg ttaattaagg ggatccgccc ctctccctcc 2401 ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata 2461 tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg 2521 tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt 2581 tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag 2641 cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc 2701 cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga 2761 tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg 2821 cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat 2881 gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc 2941 tttgaaaaac acgatgataa gcttgccaca accagatctc gccaccatgg tgagcaaggg 3001 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg 3061 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct 3121 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct 3181 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt 3241 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg 3301 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga 3361 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa 3421 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa 3481 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca 3541 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca 3601 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt 3661 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccctagagg 3721 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg 3781 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt 3841 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg 3901 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc 3961 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc 4021 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc 4081 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc 4141 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg 4201 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc 4261 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggta 4321 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag 4381 gggggactgg aagggctaat tcactcccaa cgaagataag atctgctttt tgcttgtact 4441 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca 4501 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg 4561 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc 4621 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca 4681 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 4741 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 4801 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc 4861 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt 4921 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt 4981 ttggaggcct agacttttgc agagaccaaa ttcgtaatca tgtcatagct gtttcctgtg 5041 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 5101 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 5161 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 5221 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 5281 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 5341 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 5401 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 5461 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 5521 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 5581 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 5641 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 5701 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 5761 tcgccactgg cagcagccac tggtaacagg attggcagag cgaggtatgt aggcggtgct 5821 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc 5881 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 5941 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 6001 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 6061 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 6121 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 6181 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 6241 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 6301 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 6361 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 6421 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 6481 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 6541 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 6601 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 6661 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 6721 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 6781 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 6841 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 6901 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 6961 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 7021 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 7081 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 7141 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg 7201 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat 7261 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg 7321 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc 7381 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa 7441 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg 7501 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa 7561 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt 7621 tgtaaaacga cggccagtgc caagctg //