LOCUS Exported 7507 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS LV-GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7507) 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 7507) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7507 /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" regulatory 909..918 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 915..1634 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 915..1631 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(960..981) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1221..1240) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1568..1589 /label=EGFP-C /note="EGFP, forward primer" LTR 1713..1946 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 2018..2139 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2055..2074) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2109..2128 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 2179..2314 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2241..2260 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(2334..2353) /label=T7 /note="T7 promoter, forward primer" promoter complement(2335..2353) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2363..2380) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2363..2379) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2372..2394) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 2521..2976 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2608..2627) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2818..2839 /label=F1ori-F /note="F1 origin, forward primer" promoter 3002..3106 /gene="bla" /label=AmpR promoter CDS 3107..3967 /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(3325..3344) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4138..4726 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4627..4646 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4880..4897 /label=L4440 /note="L4440 vector, forward primer" protein_bind 5014..5035 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5050..5080 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5088..5104 /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 5093..5115 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 5112..5128 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5112..5128 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 5147..5167 /label=T3 /note="T3 promoter, forward primer" promoter 5149..5167 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 5195..5421 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 5422..5602 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 5649..5774 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 6267..6500 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 6685..6729 /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 7027..7267 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 7027..7047 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 7198..7217 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_feature 7338..7455 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 7504..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 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc 961 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg 1021 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg 1081 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc 1141 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg 1201 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg 1261 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca 1321 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg 1381 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca 1441 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc 1501 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc 1561 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg 1621 agctgtacaa gtaaatgcat cggtaccttt aagaccaatg acttacaagg cagctgtaga 1681 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag 1741 acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga 1801 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct 1861 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt 1921 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt 1981 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa 2041 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca 2101 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc 2161 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc 2221 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta 2281 ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata 2341 gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc 2401 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata 2461 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg 2521 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg 2581 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca 2641 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta 2701 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc 2761 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg 2821 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat 2881 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta 2941 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt 3001 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag 3061 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca 3121 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc 3181 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat 3241 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc 3301 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg 3361 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc 3421 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat 3481 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga 3541 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc 3601 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc 3661 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt 3721 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc 3781 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc 3841 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca 3901 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca 3961 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt 4021 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta 4081 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg 4141 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc 4201 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag 4261 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa 4321 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc 4381 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc 4441 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta 4501 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag 4561 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct 4621 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga 4681 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc 4741 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt 4801 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg 4861 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg 4921 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc 4981 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc 5041 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata 5101 acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca 5161 ctaaagggaa caaaagctgg agctgcaagc ttaatgtagt cttatgcaat actcttgtag 5221 tcttgcaaca tggtaacgat gagttagcaa catgccttac aaggagagaa aaagcaccgt 5281 gcatgccgat tggtggaagt aaggtggtac gatcgtgcct tattaggaag gcaacagacg 5341 ggtctgacat ggattggacg aaccactgaa ttgccgcatt gcagagatat tgtatttaag 5401 tgcctagctc gatacataaa cgggtctctc tggttagacc agatctgagc ctgggagctc 5461 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa 5521 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag 5581 tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac 5641 cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg 5701 ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg 5761 ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt 5821 taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc 5881 tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac 5941 tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata 6001 cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt 6061 tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg 6121 atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat 6181 aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg 6241 cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca 6301 ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct 6361 ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg 6421 caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac 6481 ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact 6541 gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg 6601 acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt 6661 gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg 6721 gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa attattcata 6781 atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat 6841 agagttaggc agggatattc accattatcg tttcagaccc acctcccaac cccgagggga 6901 cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt 6961 cgattagtga acggatctcg acggtatcga tcacgagact agcctcgagc ggccgccccc 7021 ttcaccgagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt 7081 agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga 7141 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac 7201 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg 7261 aaaggacgaa acaccggtcc gcaggtatgc acgcgtgaat tctcgacctc gagacaaatg 7321 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa 7381 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca 7441 aaaattcaaa attttcgggt ttattacagg gacagcagag atccactttg gccgcggctc 7501 gaggggg //