LOCUS Exported 9034 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Lentiviral vector of luciferase-eGFP fusion gene driven by Pol2 promoter. ACCESSION . VERSION . KEYWORDS pFUGW-Pol2-ffLuc2-eGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9034) AUTHORS Day CP, Carter J, Bonomi C, Esposito D, Crise B, Ortiz-Conde B, Hollingshead M, Merlino G TITLE Lentivirus-mediated bifunctional cell labeling for in vivo melanoma study. JOURNAL Pigment Cell Melanoma Res. 2009 Jun;22(3):283-95. doi: 10.1111/j.1755-148X.2009.00545.x. Epub 2009 Jan 19. PUBMED 19175523 REFERENCE 2 (bases 1 to 9034) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9034 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(47..66) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 238..617 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 619..817 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 772..792 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 835..1015 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1062..1187 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1686..1919 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2104..2148 /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 2446..2563 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" protein_bind 2642..2662 /gene="mutant version of attB" /label=attB4 /bound_moiety="BP Clonase(TM)" /note="core recombination site for the Gateway(R) BP reaction" protein_bind 3375..3399 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 3402..5051 /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" regulatory 5067..5076 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 5073..5789 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(5118..5139) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(5379..5398) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 5726..5747 /label=EGFP-C /note="EGFP, forward primer" protein_bind complement(5794..5818) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" primer_bind 5832..5848 /label=KS primer /note="common sequencing primer, one of multiple similar variants" misc_feature 5923..6511 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5976..5996) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6394..6405) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(6514..6530) /label=KS primer /note="common sequencing primer, one of multiple similar variants" LTR 7040..7220 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" rep_origin complement(7282..7870) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7362..7381) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8041..8901) /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 8664..8683 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8902..9006) /gene="bla" /label=AmpR promoter ORIGIN 1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg 61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt 121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc 181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac 241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat 301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg 361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt 421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag 481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc 541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag 601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt 661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc 721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg 781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct 841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc 1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc 1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa 1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg 1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata 1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc 1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga 1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc 1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca 1501 aaagtaagac caccgcacag caagcggccg gccgcgctga tcttcagacc tggaggagga 1561 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca 1621 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg 1681 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg 1741 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac 1801 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc 1861 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg 1921 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt 1981 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga 2041 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa 2101 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt 2161 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta 2221 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca 2281 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata 2341 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatcggca 2401 ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg 2461 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca 2521 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga 2581 cagcagagat ccagtttggt tagtaccggg cccgctctag attgaattca agggatcatc 2641 acaactttgt atagaaaagt tggcaagctt gggcttgaac atcgagcgcc agggctccgt 2701 aaagctacta gagcacaggc ggtgccccaa cgtcctgggg cctctccact aataacggct 2761 acttccaatt gattggacgc gccatcttgc ctgccttatg catattcagc ggtgaactga 2821 atattcatga acgaggcccg tcccgtccct ccctccttcc ccccaccccc ggaacccgct 2881 ccggaggacc cgaagggccc cgccttcatt accgatgcgt aggacaaacc attttcccga 2941 tgtgtgtggg gggatactaa tgagagactt tagctgaaaa tgagcctgaa ctccgaagct 3001 gagtaaaaat ggcctaactt tatcctccgt tctgtaagtc ctcggtttga gtgcacggga 3061 aacccgaaag gaggacgaca ggaccaggaa cattctcctc ctcctgtcgc gtcagaaaga 3121 acacccaacc agggagccgg agccctagcg tcaacaactc cgccgcgcgc gctccgtgta 3181 ggccggtgcg ggcggccccg tagcgcaagg gagggcggga aaggaagggg cgggacacaa 3241 gggcgaatct ataaagggcg tcactcagcc agttctctcc tcagaagcgc cgagagcgcg 3301 accgggacgg ttggagaaga aggtggctcc cggaaggggg agagacaaac tgccgtaacc 3361 tctgccgttc aggatcaact ttgtacaaaa aagttggcac catggaagat gccaaaaaca 3421 ttaagaaggg cccagcgcca ttctacccac tcgaagacgg gaccgccggc gagcagctgc 3481 acaaagccat gaagcgctac gccctggtgc ccggcaccat cgcctttacc gacgcacata 3541 tcgaggtgga cattacctac gccgagtact tcgagatgag cgttcggctg gcagaagcta 3601 tgaagcgcta tgggctgaat acaaaccatc ggatcgtggt gtgcagcgag aatagcttgc 3661 agttcttcat gcccgtgttg ggtgccctgt tcatcggtgt ggctgtggcc ccagctaacg 3721 acatctacaa cgagcgcgag ctgctgaaca gcatgggcat cagccagccc accgtcgtat 3781 tcgtgagcaa gaaagggctg caaaagatcc tcaacgtgca aaagaagcta ccgatcatac 3841 aaaagatcat catcatggat agcaagaccg actaccaggg cttccaaagc atgtacacct 3901 tcgtgacttc ccatttgcca cccggcttca acgagtacga cttcgtgccc gagagcttcg 3961 accgggacaa aaccatcgcc ctgatcatga acagtagtgg cagtaccgga ttgcccaagg 4021 gcgtagccct accgcaccgc accgcttgtg tccgattcag tcatgcccgc gaccccatct 4081 tcggcaacca gatcatcccc gacaccgcta tcctcagcgt ggtgccattt caccacggct 4141 tcggcatgtt caccacgctg ggctacttga tctgcggctt tcgggtcgtg ctcatgtacc 4201 gcttcgagga ggagctattc ttgcgcagct tgcaagacta taagattcaa tctgccctgc 4261 tggtgcccac actatttagc ttcttcgcta agagcactct catcgacaag tacgacctaa 4321 gcaacttgca cgagatcgcc agcggcgggg cgccgctcag caaggaggta ggtgaggccg 4381 tggccaaacg cttccaccta ccaggcatcc gccagggcta cggcctgaca gaaacaacca 4441 gcgccattct gatcaccccc gaaggggacg acaagcctgg cgcagtaggc aaggtggtgc 4501 ccttcttcga ggctaaggtg gtggacttgg acaccggtaa gacactgggt gtgaaccagc 4561 gcggcgagct gtgcgtccgt ggccccatga tcatgagcgg ctacgttaac aaccccgagg 4621 ctacaaacgc tctcatcgac aaggacggct ggctgcacag cggcgacatc gcctactggg 4681 acgaggacga gcacttcttc atcgtggacc ggctgaagag cctgatcaaa tacaagggct 4741 accaggtagc cccagccgaa ctggagagca tcctgctgca acaccccaac atcttcgacg 4801 ccggggtcgc cggcctgccc gacgacgatg ccggcgagct gcccgccgca gtcgtcgtgc 4861 tggaacacgg taaaaccatg accgagaagg agatcgtgga ctatgtggcc agccaggtta 4921 caaccgccaa gaagctgcgc ggtggtgttg tgttcgtgga cgaggtgcct aaaggactga 4981 ccggcaagtt ggacgcccgc aagatccgcg agattctcat taaggccaag aagggcggca 5041 agatcgccgt gcgggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca 5101 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg 5161 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca 5221 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc 5281 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc 5341 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc 5401 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg 5461 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca 5521 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc 5581 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg 5641 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca 5701 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga 5761 tcactctcgg catggacgag ctgtacaagt agtaaccaac tttcttgtac aaagttgtga 5821 tgatccaatc tcgaggtcga cggtatcgat aagctcgctt cacgagatca tgtttaaggg 5881 ttccggttcc actaggtaca attcgatatc aagcttatcg ataatcaacc tctggattac 5941 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 6001 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 6061 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 6121 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 6181 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc 6241 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 6301 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg 6361 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 6421 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg 6481 agtcggatct ccctttgggc cgcctccccg catcgatacc gtcgacctcg atcgagacct 6541 agaaaaacat ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg 6601 gctagaagca caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag 6661 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact 6721 ggaagggcta attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac 6781 acaaggctac ttccctgatt ggcagaacta cacaccaggg ccagggatca gatatccact 6841 gacctttgga tggtgctaca agctagtacc agttgagcaa gagaaggtag aagaagccaa 6901 tgaaggagag aacacccgct tgttacaccc tgtgagcctg catgggatgg atgacccgga 6961 gagagaagta ttagagtgga ggtttgacag ccgcctagca tttcatcaca tggcccgaga 7021 gctgcatccg gactgtactg ggtctctctg gttagaccag atctgagcct gggagctctc 7081 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt 7141 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc 7201 agtgtggaaa atctctagca gcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 7261 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 7321 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 7381 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 7441 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 7501 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 7561 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 7621 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 7681 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg 7741 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 7801 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 7861 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 7921 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 7981 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 8041 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 8101 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 8161 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 8221 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 8281 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 8341 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 8401 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 8461 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 8521 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 8581 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 8641 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 8701 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 8761 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 8821 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 8881 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 8941 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 9001 tccgcgcaca tttccccgaa aagtgccacc tgac //