LOCUS Exported 7760 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Lentiviral plasmid for expression of far red iRFP720 nuclear marker. ACCESSION . VERSION . KEYWORDS pLenti-H2B-iRFP720 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7760) TITLE Nuclear fluorescent proteins JOURNAL Unpublished REFERENCE 2 (bases 1 to 7760) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7760 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 426..444 /label=hPGK-F /note="Human PGK promoter, forward primer" CDS 519..896 /codon_start=1 /gene="HIST1H2BJ" /product="human histone H2B" /label=H2B /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP GELAKHAVSEGTKAITKYTSAK" CDS 915..1862 /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="AEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGSD NLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFIG SWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKITG FDRVMIYRFASDFSGSVIAEDRCAEVESKLGLHYPASFIPAQARRLYTINPVRIIPDIN YRPVPVTPDLNPVTGRPIDLSFAILRSVSPNHLEFMRNIGMHGTMSISILRGERLWGLI VCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE" LTR 1933..2166 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 2238..2359 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2275..2294) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2329..2348 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 2399..2534 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2461..2480 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(2554..2573) /label=T7 /note="T7 promoter, forward primer" promoter complement(2555..2573) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2583..2600) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2583..2599) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2592..2614) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 2741..3196 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2828..2847) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3038..3059 /label=F1ori-F /note="F1 origin, forward primer" promoter 3222..3326 /gene="bla" /label=AmpR promoter CDS 3327..4187 /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 complement(3545..3564) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4358..4946 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4847..4866 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5100..5117 /label=L4440 /note="L4440 vector, forward primer" protein_bind 5234..5255 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5270..5300 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5308..5324 /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)." primer_bind 5313..5335 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 5332..5348 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5332..5348 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 5367..5387 /label=T3 /note="T3 promoter, forward primer" promoter 5369..5387 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 5415..5641 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 5642..5822 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 5869..5994 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 6487..6720 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 6905..6949 /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" promoter 7247..7487 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 7247..7267 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 7418..7437 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_feature 7591..7708 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 7757..507 /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" ORIGIN 1 ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg 61 cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt 121 cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct 181 ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa 241 gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc 301 cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc 361 cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc 421 gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt 481 gaccgaatca ccgacctctc tccccagggg gatctaccat gccagagcca gcgaagtctg 541 ctcccgcccc gaaaaagggc tccaagaagg cggtgactaa ggcgcagaag aaaggcggca 601 agaagcgcaa gcgcagccgc aaggagagct attccatcta tgtgtacaag gttctgaagc 661 aggtccaccc tgacaccggc atttcgtcca aggccatggg catcatgaac tcgtttgtga 721 acgacatttt cgagcgcatc gcaggtgagg cttcccgcct ggcgcattac aacaagcgct 781 cgaccatcac ctccagggag atccagacgg ccgtgcgcct gctgctgcct ggggagttgg 841 ccaagcacgc cgtgtccgag ggtactaagg ccatcaccaa gtacaccagc gctaaggatc 901 caccggtcgc caccgcggaa ggatccgtcg ccaggcagcc tgacctcttg acctgcgacg 961 atgagccgat ccatatcccc ggtgccatcc aaccgcatgg actgctgctc gccctcgccg 1021 ccgacatgac gatcgttgcc ggcagcgaca accttcccga actcaccgga ctggcgatcg 1081 gcgccctgat cggccgctct gcggccgatg tcttcgactc ggagacgcac aaccgtctga 1141 cgatcgcctt ggccgagccc ggggcggccg tcggagcacc gatcactgtc ggcttcacga 1201 tgcgaaagga cgcaggcttc atcggctcct ggcatcgcca tgatcagctc atcttcctcg 1261 agctcgagcc tccccagcgg gacgtcgccg agccgcaggc gttcttccgc cgcaccaaca 1321 gcgccatccg ccgcctgcag gccgccgaaa ccttggaaag cgcctgcgcc gccgcggcgc 1381 aagaggtgcg gaagattacc ggcttcgatc gggtgatgat ctatcgcttc gcctccgact 1441 tcagcgggtc cgtgatcgca gaggatcggt gcgccgaggt cgagtcaaaa ctaggcctgc 1501 actatcctgc ctcattcatc ccggcgcagg cccgtcggct ctataccatc aacccggtac 1561 ggatcattcc cgatatcaat tatcggccgg tgccggtcac cccagacctc aatccggtca 1621 ccgggcggcc gattgatctt agcttcgcca tcctgcgcag cgtctcgccc aaccatctgg 1681 agttcatgcg caacataggc atgcacggca cgatgtcgat ctcgattttg cgcggcgagc 1741 gactgtgggg attgatcgtt tgccatcacc gaacgccgta ctacgtcgat ctcgatggcc 1801 gccaagcctg cgagctagtc gcccaggttc tggcctggca gatcggcgtg atggaagagt 1861 aagtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag 1921 aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg ctttttgctt 1981 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga 2041 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc 2101 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct 2161 ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa 2221 tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata 2281 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca 2341 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccat 2401 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt 2461 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg 2521 cttttttgga ggcctaggga cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc 2581 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat 2641 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat 2701 cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca 2761 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 2821 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 2881 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac 2941 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt 3001 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 3061 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg 3121 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata 3181 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt 3241 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc 3301 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc 3361 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa 3421 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg 3481 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag 3541 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc 3601 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta 3661 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg 3721 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca 3781 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac 3841 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat 3901 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg 3961 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata 4021 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta 4081 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa 4141 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag 4201 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg 4261 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact 4321 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg 4381 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc 4441 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata 4501 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta 4561 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc 4621 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg 4681 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac 4741 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg 4801 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt 4861 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct 4921 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg 4981 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata 5041 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca 5101 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc 5161 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg 5221 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta 5281 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca 5341 gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg 5401 agctgcaagc ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat 5461 gagttagcaa catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt 5521 aaggtggtac gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg 5581 aaccactgaa ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa 5641 cgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc 5701 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt 5761 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag 5821 cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc 5881 aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg 5941 ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt 6001 aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa 6061 aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat 6121 cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc 6181 cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt 6241 gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag 6301 caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg 6361 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc 6421 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt 6481 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc 6541 gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa 6601 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat 6661 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct 6721 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag 6781 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag 6841 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca 6901 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt 6961 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt 7021 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc 7081 accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat 7141 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatctcg 7201 acggtatcga tcacgagact agcctcgagc ggccgccccc ttcaccgagg gcctatttcc 7261 catgattcct tcatatttgc atatacgata caaggctgtt agagagataa ttggaattaa 7321 tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct 7381 tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac 7441 ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccggcaa 7501 caagatgaag agcaccaact cgagttggtg ctcttcatct tgttgttttt aattctcgac 7561 ctcgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta 7621 cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca 7681 aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca gagatccact 7741 ttggccgcgg ctcgaggggg //