LOCUS Cellecta-pRSG16- 8025 bp ds-DNA circular SYN 12-JUL-2018 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8025) AUTHORS Cellecta TITLE Direct Submission JOURNAL Exported Jul 12, 2018 from SnapGene 4.2.0 to Vector NTI(R) format http://www.snapgene.com COMMENT VNTDATE|662774400| COMMENT VNTDBDATE|847065600| COMMENT VNTNAME|Cellecta-pRSG16-U6-sg-HTS6C-UbiC-TagRFP-2A-Puro.dna| COMMENT VNTAUTHORNAME|Cellecta| FEATURES Location/Qualifiers promoter 6..232 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 233..413 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 460..585 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1078..1311 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." promoter 1819..2059 /label=U6 /note="RNA polymerase III promoter for human U6 snRNA" primer 1863..1888 /label=FwdU6-1 /note="sequence: caaggctgttagagagataattggaa" primer 1914..1939 /label=FwdU6-3 /note="sequence: attagtacaaaatacgtgacgtagaa" primer 1987..2017 /label=P5-NFG16 /note="sequence: AGATACGGCGACCACCGAGATCTACACGAGATGGACTATCATATGCTTACCGTAACTTG AA" primer 2034..2068 /label=FSeq-gRNA /note="sequence: GGCTTTATATATCTTGTGGAAAGGACGAAACACCG" misc_feature 2065..2094 /label=cloning site misc_feature 2122..2237 /label=cPPT /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" primer complement(2169..2195) /label=P7-NRG16 /note="sequence: CAAGCAGAAGACGGCATACGAGATGCACGACGAGACGCAGACGAATACGACAGAGTGTA TGTCTGTTGCTATTATGTCTACT" primer complement(2227..2251) /label=Rev-1 /note="sequence: cgacaacaacgcagaaattttgaat" promoter 2303..2702 /label=UbiC /note="human ubiquitin C promoter" CDS 2715..3425 /codon_start=1 /product="monomeric derivative of red fluorescent protein from Entacmaea quadricolor (Merzlyak et al., 2007)" /label=TagRFP /note="mammalian codon-optimized" CDS 3432..3485 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." CDS 3492..4091 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" misc_feature 4104..4692 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" LTR 4766..4999 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 5071..5192 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin 5232..5367 /label=SV40 ori /note="SV40 origin of replication" rep_origin complement(5802..6390) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6561..7421) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" promoter complement(7422..7526) /gene="bla" /label=AmpR promoter ORIGIN 1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca 61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta 121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga 181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacataa acgggtctct 241 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa 301 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc 361 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc 421 ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg 481 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt 541 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg 601 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa 661 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg 721 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca 781 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa 841 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa 901 agtaagacca ccgcacagca agcggccact gatcttcaga cctggaggag gagatatgag 961 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt 1021 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg 1081 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac 1141 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct 1201 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct 1261 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg 1321 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa 1381 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa 1441 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa 1501 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac 1561 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag 1621 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc 1681 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga 1741 aggtggagag agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg 1801 atggccgccc ccttcaccga gggcctattt cccatgattc cttcatattt gcatatacga 1861 tacaaggctg ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta 1921 caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt 1981 tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta 2041 tatatcttgt ggaaaggacg aaacaccgga gtcttctttt ttgaagacac ttcggactgt 2101 agaactctga acctcgagca atttaaaaga aaagggggga ttggggggta cagtgcaggg 2161 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt 2221 acaaaaattc aaaatttctg cgttgttgtc ggtgctcgtt ctctgctctt cacgctactg 2281 aattcatcac cggttcttcg aaggcctccg cgccgggttt tggcgcctcc cgcgggcgcc 2341 cccctcctca cggcgagcgc tgccacgtca gacgaagggc gcagcgagcg tcctgatcct 2401 tccgcccgga cgctcaggac agcggcccgc tgctcataag actcggcctt agaaccccag 2461 tatcagcaga aggacatttt aggacgggac ttgggtgact ctagggcact ggttttcttt 2521 ccagagagcg gaacaggcga ggaaaagtag tcccttctcg gcgattctgc ggagggatct 2581 ccgtggggcg gtgaacgccg atgattatat aaggacgcgc cgggtgtggc acagctagtt 2641 ccgtcgcagc cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg 2701 gtctagacgc caccatggtg tctaagggcg aagagctgat taaggagaac atgcacatga 2761 agctgtacat ggagggcacc gtgaacaacc accacttcaa gtgcacatcc gagggcgaag 2821 gcaagcccta cgagggcacc cagaccatga gaatcaaggt ggtcgagggc ggccctctcc 2881 ccttcgcctt cgacatcctg gctaccagct tcatgtacgg cagcagaacc ttcatcaacc 2941 acacccaggg catccccgac ttctttaagc agtccttccc tgagggcttc acatgggaga 3001 gagtcaccac atacgaggac gggggcgtgc tgaccgctac ccaggacacc agcctccagg 3061 acggctgcct catctacaac gtcaagatca gaggggtgaa cttcccatcc aacggccctg 3121 tgatgcagaa gaaaacactc ggctgggagg ccaacaccga gatgctgtac cccgctgacg 3181 gcggcctgga aggcagaagc gacatggccc tgaagctcgt gggcgggggc cacctgatct 3241 gcaacttcaa gaccacatac agatccaaga aacccgctaa gaacctcaag atgcccggcg 3301 tctactatgt ggaccacaga ctggaaagaa tcaaggaggc cgacaaagag acctacgtcg 3361 agcagcacga ggtggctgtg gccagatact gcgacctccc tagcaaactg gggcacaaac 3421 ttaatggatc cgagggcaga ggaagccttc taacatgcgg tgacgtggag gagaatcccg 3481 gcccttccgg gatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc 3541 ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg 3601 tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg 3661 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga 3721 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg 3781 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc 3841 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca 3901 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc 3961 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca 4021 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc 4081 ccggtgcctg aatctaggtc gacaatcaac ctctggatta caaaatttgt gaaagattga 4141 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt 4201 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt 4261 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg 4321 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg 4381 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc 4441 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat 4501 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct 4561 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg 4621 ctctgcggcc tcttccgcgt ctccgccttc gccctcagac gagtcggatc tctctttggg 4681 ccgcctcccc gcctggtacc tttaagacca atgacttaca aggcagctgt agatcttagc 4741 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagataagat 4801 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct 4861 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta 4921 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca 4981 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt 5041 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac 5101 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt 5161 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc 5221 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct 5281 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga 5341 agtagtgagg aggctttttt ggaggcctag acttttgcag agaccaaatt cgtaatcatg 5401 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc 5461 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg 5521 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc 5581 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact 5641 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta 5701 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag 5761 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc 5821 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta 5881 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg 5941 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc 6001 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac 6061 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac 6121 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg 6181 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga 6241 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt 6301 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag 6361 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 6421 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 6481 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat 6541 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc 6601 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg 6661 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct 6721 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca 6781 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg 6841 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg 6901 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc 6961 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag 7021 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg 7081 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag 7141 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat 7201 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg 7261 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca 7321 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca 7381 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat 7441 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag 7501 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa 7561 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt 7621 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc 7681 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt 7741 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg 7801 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc 7861 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta 7921 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg 7981 ttttcccagt cacgacgttg taaaacgacg gccagtgcca agctg //