LOCUS Exported 7539 bp ds-DNA circular SYN 10-JAN-2023 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS LV-RFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7539) AUTHORS Beronja S, Livshits G, Williams S, Fuchs E TITLE Rapid functional dissection of genetic networks via tissue-specific transduction and RNAi in mouse embryos. JOURNAL Nat Med. 2010 Jul . 16(7):821-7. PUBMED 20526348 REFERENCE 2 (bases 1 to 7539) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 10, 2023 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7539 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..511 /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" primer_bind 430..448 /label=hPGK-F /note="Human PGK promoter, forward primer" CDS 523..900 /codon_start=1 /gene="HIST1H2BJ" /product="human histone H2B" /label=H2B /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP GELAKHAVSEGTKAITKYTSAK" regulatory 913..922 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 919..1596 /codon_start=1 /product="monomeric derivative of DsRed (Campbell et al., 2002)" /label=mRFP1 /note="mammalian codon-optimized" /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS TGA" primer_bind complement(1049..1067) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1317..1336 /label=mCherry-F /note="mCherry, forward primer" primer_bind 1511..1534 /label=DsRed1-C /note="DsRed1, forward primer" LTR 1716..1949 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 2021..2142 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2058..2077) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2112..2131 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 2182..2317 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2244..2263 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(2337..2356) /label=T7 /note="T7 promoter, forward primer" promoter complement(2338..2356) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2366..2383) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2366..2382) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2375..2397) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 2524..2979 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2611..2630) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2821..2842 /label=F1ori-F /note="F1 origin, forward primer" promoter 3005..3109 /gene="bla" /label=AmpR promoter CDS 3110..3970 /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(3328..3347) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4141..4729 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4630..4649 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4883..4900 /label=L4440 /note="L4440 vector, forward primer" protein_bind 5017..5038 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5053..5083 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5091..5107 /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 5096..5118 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 5115..5131 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5115..5131 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 5150..5170 /label=T3 /note="T3 promoter, forward primer" promoter 5152..5170 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 5198..5424 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 5425..5605 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 5652..5777 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 6270..6503 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 6688..6732 /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 7030..7270 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 7030..7050 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 7201..7220 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_feature 7374..7491 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" ORIGIN 1 ggggttgggg ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc 61 tgggcgtggt tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc 121 cgttcgcagc gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc 181 tgctccgccc ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac 241 ggaagccgca cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc 301 gcgccgaccg cgatgggctg tggccaatag cggctgctca gcagggcgcg ccgagagcag 361 cggccgggaa ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct 421 gcccgcgcgg tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct 481 cgttgaccga atcaccgacc tctctcccca gggggatcta ccatgccaga gccagcgaag 541 tctgctcccg ccccgaaaaa gggctccaag aaggcggtga ctaaggcgca gaagaaaggc 601 ggcaagaagc gcaagcgcag ccgcaaggag agctattcca tctatgtgta caaggttctg 661 aagcaggtcc accctgacac cggcatttcg tccaaggcca tgggcatcat gaactcgttt 721 gtgaacgaca ttttcgagcg catcgcaggt gaggcttccc gcctggcgca ttacaacaag 781 cgctcgacca tcacctccag ggagatccag acggccgtgc gcctgctgct gcctggggag 841 ttggccaagc acgccgtgtc cgagggtact aaggccatca ccaagtacac cagcgctaag 901 gatccaccgg tcgccaccat ggcctcctcc gaggacgtca tcaaggagtt catgcgcttc 961 aaggtgcgca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag 1021 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg cggccccctg 1081 cccttcgcct gggacatcct gtcccctcag ttccagtacg gctccaaggc ctacgtgaag 1141 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag 1201 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag 1261 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc 1321 gtaatgcaga agaagaccat gggctgggag gcctccaccg agcggatgta ccccgaggac 1381 ggcgccctga agggcgagat caagatgagg ctgaagctga aggacggcgg ccactacgac 1441 gccgaggtca agaccaccta catggccaag aagcccgtgc agctgcccgg cgcctacaag 1501 accgacatca agctggacat cacctcccac aacgaggact acaccatcgt ggaacagtac 1561 gagcgcgccg agggccgcca ctccaccggc gcctaactcg agactagtcg acgtgaggat 1621 cctctagagt ccctaggatg catcggtacc tttaagacca atgacttaca aggcagctgt 1681 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg 1741 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg 1801 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt 1861 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc 1921 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta 1981 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta 2041 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 2101 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct 2161 atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg 2221 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag 2281 ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct 2341 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa 2401 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta 2461 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat 2521 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga 2581 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg 2641 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat 2701 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg 2761 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata 2821 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt 2881 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat 2941 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa 3001 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat 3061 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca 3121 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca 3181 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta 3241 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt 3301 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc 3361 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc 3421 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc 3481 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa 3541 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga 3601 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat 3661 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca 3721 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc 3781 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat 3841 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag 3901 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa 3961 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca 4021 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc 4081 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc 4141 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 4201 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 4261 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt 4321 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 4381 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 4441 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 4501 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 4561 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 4621 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 4681 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 4741 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc 4801 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg 4861 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat 4921 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt 4981 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta 5041 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg 5101 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc 5161 tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg 5221 tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac 5281 cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag 5341 acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt 5401 aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag 5461 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt 5521 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt 5581 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga 5641 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg 5701 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag 5761 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc 5821 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg 5881 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa 5941 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata 6001 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag 6061 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg 6121 ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa 6181 tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg 6241 gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca 6301 gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg 6361 tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg 6421 ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga 6481 tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc 6541 actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac 6601 acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta 6661 attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa 6721 tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc 6781 ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg 6841 aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg 6901 ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc 6961 attcgattag tgaacggatc tcgacggtat cgatcacgag actagcctcg agcggccgcc 7021 cccttcaccg agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct 7081 gttagagaga taattggaat taatttgact gtaaacacaa agatattagt acaaaatacg 7141 tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg 7201 gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg 7261 tggaaaggac gaaacaccgg caacaagatg aagagcacca actcgagttg gtgctcttca 7321 tcttgttgtt tttaattctc gacctcgaga caaatggcag tattcatcca caattttaaa 7381 agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca 7441 gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat 7501 tacagggaca gcagagatcc actttggccg cggctcgag //