pHIV-iRFP720-E2A-Luc

LOCUS       pHIV-iRFP720-E2A-Luc                9050 bp ds-DNA     circular SYN 06-AUG-2018
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 Aug 6, 2018 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."
     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
//