LOCUS Exported 9050 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Lentivirus vector for bi-cistronic expression of iRFP720 and firefly luciferase from EF1a promoter. ACCESSION . VERSION . KEYWORDS pHIV-iRFP720-E2A-Luc SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9050) AUTHORS Comenge J, Sharkey J, Fragueiro O, Wilm B, Brust M, Murray P, Levy R, Plagge A TITLE Multimodal cell tracking from systemic administration to tumour growth by combining gold nanorods and reporter genes. JOURNAL Elife. 2018 Jun 27;7. pii: 33140. doi: 10.7554/eLife.33140. PUBMED 29949503 REFERENCE 2 (bases 1 to 9050) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9050 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 414..433 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(652..756) /gene="bla" /label=AmpR promoter primer_bind complement(831..850) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 1022..1401 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1403..1601 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 1556..1576 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 1619..1799 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1846..1971 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 2468..2701 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2886..2930 /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 3228..3345 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 3408..4586 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 3639..4577 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 4534..4554 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" regulatory 4645..4654 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 4651..5598 /codon_start=1 /product="near-infrared fluorescent protein with an emission peak at 720 nm, engineered from a bacterial phytochrome (Shcherbakova and Verkhusha, 2013)" /label=iRFP720 /note="derived from Rhodopseudomonas palustris RpBphP2" /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT GFDRVMIYRFASDFSGSVIAEDRCAEVESKLGLHYPASFIPAQARRLYTINPVRIIPDI NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPNHLEFMRNIGMHGTMSISILRGERLWGL IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE" CDS 5599..5658 /codon_start=1 /product="2A peptide from equine rhinitis A virus polyprotein" /label=E2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="QCTNYALLKLAGDVESNPGP" CDS 5659..7311 /codon_start=1 /product="firefly luciferase" /label=luciferase /note="synthetic luc2 version of the luciferase gene" /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV" primer_bind complement(7314..7330) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7315..7331) /label=pBluescriptKS /note="For pBluescript vector" LTR 7840..8020 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" rep_origin complement(8082..8670) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(8162..8181) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8841..651) /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" ORIGIN 1 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt 61 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt 121 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt 181 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt 241 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct 301 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt 361 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac 421 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa 481 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa 541 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca 601 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct 661 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga 721 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc 781 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc 841 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag 901 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag 961 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt 1021 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc 1081 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc 1141 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg 1201 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat 1261 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc 1321 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta 1381 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag 1441 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt 1501 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa 1561 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg cctgtactgg 1621 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact 1681 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg 1741 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag 1801 tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg 1861 actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca 1921 aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag 1981 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa 2041 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct 2101 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt 2161 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg 2221 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa 2281 aacaaaagta agaccaccgc acagcaagcg gccggccgct gatcttcaga cctggaggag 2341 gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac 2401 cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag 2461 tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag 2521 cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga 2581 acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca 2641 tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc 2701 tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta 2761 gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca 2821 gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc 2881 aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt 2941 ttaacataac aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg 3001 taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt 3061 caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa 3121 tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg aacggatcgg 3181 cactgcgtgc gccaattctg cagacaaatg gcagtattca tccacaattt taaaagaaaa 3241 ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata 3301 caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg 3361 gacagcagag atccagtttg gttagtaccg ggcccgctct agcgtgaggc tccggtgccc 3421 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca 3481 attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact 3541 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga 3601 acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg tgtggttccc 3661 gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacctggc 3721 tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc 3781 cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg 3841 gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc 3901 tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg 3961 taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg 4021 gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga 4081 gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc 4141 cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg 4201 aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg cggcgctcgg 4261 gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg 4321 cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct 4381 tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca 4441 ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat 4501 ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt 4561 tttttcttcc atttcaggtg tcgtgagcgg ccgctgaact gaattctgca gtcgacggta 4621 ccgcgggccc gggatccacc ggtcgccacc atggcggaag gatccgtcgc caggcagcct 4681 gacctcttga cctgcgacga tgagccgatc catatccccg gtgccatcca accgcatgga 4741 ctgctgctcg ccctcgccgc cgacatgacg atcgttgccg gcagcgacaa ccttcccgaa 4801 ctcaccggac tggcgatcgg cgccctgatc ggccgctctg cggccgatgt cttcgactcg 4861 gagacgcaca accgtctgac gatcgccttg gccgagcccg gggcggccgt cggagcaccg 4921 atcactgtcg gcttcacgat gcgaaaggac gcaggcttca tcggctcctg gcatcgccat 4981 gatcagctca tcttcctcga gctcgagcct ccccagcggg acgtcgccga gccgcaggcg 5041 ttcttccgcc gcaccaacag cgccatccgc cgcctgcagg ccgccgaaac cttggaaagc 5101 gcctgcgccg ccgcggcgca agaggtgcgg aagattaccg gcttcgatcg ggtgatgatc 5161 tatcgcttcg cctccgactt cagcgggtcc gtgatcgcag aggatcggtg cgccgaggtc 5221 gagtcaaaac taggcctgca ctatcctgcc tcattcatcc cggcgcaggc ccgtcggctc 5281 tataccatca acccggtacg gatcattccc gatatcaatt atcggccggt gccggtcacc 5341 ccagacctca atccggtcac cgggcggccg attgatctta gcttcgccat cctgcgcagc 5401 gtctcgccca accatctgga gttcatgcgc aacataggca tgcacggcac gatgtcgatc 5461 tcgattttgc gcggcgagcg actgtgggga ttgatcgttt gccatcaccg aacgccgtac 5521 tacgtcgatc tcgatggccg ccaagcctgc gagctagtcg cccaggttct ggcctggcag 5581 atcggcgtga tggaagagca atgtacaaac tacgctttgt tgaaactcgc tggcgatgtt 5641 gaaagtaacc ccggtcctat ggaagatgcc aaaaacatta agaagggccc agcgccattc 5701 tacccactcg aagacgggac cgccggcgag cagctgcaca aagccatgaa gcgctacgcc 5761 ctggtgcccg gcaccatcgc ctttaccgac gcacatatcg aggtggacat tacctacgcc 5821 gagtacttcg agatgagcgt tcggctggca gaagctatga agcgctatgg gctgaataca 5881 aaccatcgga tcgtggtgtg cagcgagaat agcttgcagt tcttcatgcc cgtgttgggt 5941 gccctgttca tcggtgtggc tgtggcccca gctaacgaca tctacaacga gcgcgagctg 6001 ctgaacagca tgggcatcag ccagcccacc gtcgtattcg tgagcaagaa agggctgcaa 6061 aagatcctca acgtgcaaaa gaagctaccg atcatacaaa agatcatcat catggatagc 6121 aagaccgact accagggctt ccaaagcatg tacaccttcg tgacttccca tttgccaccc 6181 ggcttcaacg agtacgactt cgtgcccgag agcttcgacc gggacaaaac catcgccctg 6241 atcatgaaca gtagtggcag taccggattg cccaagggcg tagccctacc gcaccgcacc 6301 gcttgtgtcc gattcagtca tgcccgcgac cccatcttcg gcaaccagat catccccgac 6361 accgctatcc tcagcgtggt gccatttcac cacggcttcg gcatgttcac cacgctgggc 6421 tacttgatct gcggctttcg ggtcgtgctc atgtaccgct tcgaggagga gctattcttg 6481 cgcagcttgc aagactataa gattcaatct gccctgctgg tgcccacact atttagcttc 6541 ttcgctaaga gcactctcat cgacaagtac gacctaagca acttgcacga gatcgccagc 6601 ggcggggcgc cgctcagcaa ggaggtaggt gaggccgtgg ccaaacgctt ccacctacca 6661 ggcatccgcc agggctacgg cctgacagaa acaaccagcg ccattctgat cacccccgaa 6721 ggggacgaca agcctggcgc agtaggcaag gtggtgccct tcttcgaggc taaggtggtg 6781 gacttggaca ccggtaagac actgggtgtg aaccagcgcg gcgagctgtg cgtccgtggc 6841 cccatgatca tgagcggcta cgttaacaac cccgaggcta caaacgctct catcgacaag 6901 gacggctggc tgcacagcgg cgacatcgcc tactgggacg aggacgagca cttcttcatc 6961 gtggaccggc tgaagagcct gatcaaatac aagggctacc aggtagcccc agccgaactg 7021 gagagcatcc tgctgcaaca ccccaacatc ttcgacgccg gggtcgccgg cctgcccgac 7081 gacgatgccg gcgagctgcc cgccgcagtc gtcgtgctgg aacacggtaa aaccatgacc 7141 gagaaggaga tcgtggacta tgtggccagc caggttacaa ccgccaagaa gctgcgcggt 7201 ggtgttgtgt tcgtggacga ggtgcctaaa ggactgaccg gcaagttgga cgcccgcaag 7261 atccgcgaga ttctcattaa ggccaagaag ggcggcaaga tcgccgtgta aatcgatacc 7321 gtcgacctcg atcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct 7381 accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc 7441 acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt 7501 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca agatatcctt 7561 gatctgtgga tctaccacac acaaggctac ttccctgatt ggcagaacta cacaccaggg 7621 ccagggatca gatatccact gacctttgga tggtgctaca agctagtacc agttgagcaa 7681 gagaaggtag aagaagccaa tgaaggagag aacacccgct tgttacaccc tgtgagcctg 7741 catgggatgg atgacccgga gagagaagta ttagagtgga ggtttgacag ccgcctagca 7801 tttcatcaca tggcccgaga gctgcatccg gactgtactg ggtctctctg gttagaccag 7861 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc 7921 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga 7981 tccctcagac ccttttagtc agtgtggaaa atctctagca gcatgtgagc aaaaggccag 8041 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc 8101 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta 8161 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg 8221 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc 8281 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac 8341 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac 8401 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg 8461 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga 8521 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt 8581 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag 8641 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 8701 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 8761 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat 8821 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc 8881 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg 8941 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct 9001 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag //