LOCUS Cellecta-pRSG20- 7963 bp ds-DNA circular SYN 27-MAR-2017 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7963) AUTHORS Cellecta TITLE Direct Submission JOURNAL Exported Mar 27, 2017 from SnapGene 2.7.3 to Vector NTI(R) format http://www.snapgene.com COMMENT VNTDATE|753321600| COMMENT VNTDBDATE|801360000| COMMENT VNTNAME|Cellecta-pRSG20-U6-sg-HTS6C-CMV-TagRFP-2A-Puro| 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..2064 /label=U6 /note="RNA polymerase III promoter for human U6 snRNA" misc_feature 2065..2090 /label=gRNA Cloning Site misc_feature 2122..2237 /label=cPPT /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" promoter 2295..2638 /label=CMV CDS 2653..3363 /codon_start=1 /product="monomeric derivative of red fluorescent protein from Entacmaea quadricolor (Merzlyak et al., 2007)" /label=TagRFP /note="mammalian codon-optimized" CDS 3370..3423 /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 3430..4029 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" misc_feature 4042..4630 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" LTR 4704..4937 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 5009..5130 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin 5170..5305 /label=SV40 ori /note="SV40 origin of replication" rep_origin complement(5740..6328) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6499..7359) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" promoter complement(7360..7464) /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 cggtattatg cccagtacat gaccttatgg gactttccta cttggcagta 2341 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg 2401 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgccaatgg 2461 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc 2521 attgacgcaa atgggcggta ggcgtgtacg gtggaaggtt tatataagca gagctcgttt 2581 agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagatt 2641 ctagacgcca ccatggtgtc taagggcgaa gagctgatta aggagaacat gcacatgaag 2701 ctgtacatgg agggcaccgt gaacaaccac cacttcaagt gcacatccga gggcgaaggc 2761 aagccctacg agggcaccca gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc 2821 ttcgccttcg acatcctggc taccagcttc atgtacggca gcagaacctt catcaaccac 2881 acccagggca tccccgactt ctttaagcag tccttccctg agggcttcac atgggagaga 2941 gtcaccacat acgaggacgg gggcgtgctg accgctaccc aggacaccag cctccaggac 3001 ggctgcctca tctacaacgt caagatcaga ggggtgaact tcccatccaa cggccctgtg 3061 atgcagaaga aaacactcgg ctgggaggcc aacaccgaga tgctgtaccc cgctgacggc 3121 ggcctggaag gcagaagcga catggccctg aagctcgtgg gcgggggcca cctgatctgc 3181 aacttcaaga ccacatacag atccaagaaa cccgctaaga acctcaagat gcccggcgtc 3241 tactatgtgg accacagact ggaaagaatc aaggaggccg acaaagagac ctacgtcgag 3301 cagcacgagg tggctgtggc cagatactgc gacctcccta gcaaactggg gcacaaactt 3361 aatggatccg agggcagagg aagccttcta acatgcggtg acgtggagga gaatcccggc 3421 ccttccggga tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc 3481 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc 3541 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc 3601 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc 3661 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag 3721 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg 3781 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag 3841 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc 3901 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc 3961 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc 4021 ggtgcctgaa tctaggtcga caatcaacct ctggattaca aaatttgtga aagattgact 4081 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg 4141 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg 4201 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg 4261 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg 4321 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc 4381 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca 4441 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc 4501 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct 4561 ctgcggcctc ttccgcgtct ccgccttcgc cctcagacga gtcggatctc tctttgggcc 4621 gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca 4681 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gataagatct 4741 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg 4801 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt 4861 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt 4921 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc 4981 aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa 5041 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 5101 tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta 5161 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga 5221 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag 5281 tagtgaggag gcttttttgg aggcctagac ttttgcagag accaaattcg taatcatgtc 5341 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg 5401 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt 5461 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 5521 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 5581 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 5641 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 5701 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 5761 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 5821 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 5881 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc 5941 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 6001 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 6061 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 6121 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 6181 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 6241 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 6301 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 6361 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 6421 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 6481 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 6541 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 6601 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 6661 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 6721 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 6781 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 6841 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 6901 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 6961 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 7021 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 7081 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 7141 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 7201 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 7261 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 7321 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 7381 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 7441 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga 7501 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct 7561 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac 7621 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt 7681 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca 7741 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca 7801 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt 7861 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt 7921 ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctg //