LOCUS Cellecta-pRSG21- 7966 bp ds-DNA circular SYN 14-SEP-2017 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7966) AUTHORS Cellecta TITLE Direct Submission JOURNAL Exported Sep 14, 2017 from SnapGene 4.0.4 to Vector NTI(R) format http://www.snapgene.com COMMENT VNTDATE|753321600| COMMENT VNTDBDATE|816825600| COMMENT VNTNAME|Cellecta-pRSG21-U6-sg-HTS6C-CMV-TagGFP2-2A-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..2064 /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" 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(2170..2195) /label=Rev-cPPT10 /note="sequence: tgtatgtctgttgctattatgtctac" promoter 2295..2638 /label=CMV CDS 2653..3366 /codon_start=1 /product="monomeric green fluorescent protein, also known as mTagGFP" /label=TagGFP2 /note="mammalian codon-optimized" CDS 3373..3426 /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 3433..4032 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" misc_feature 4045..4633 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" LTR 4707..4940 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 5012..5133 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin 5173..5308 /label=SV40 ori /note="SV40 origin of replication" rep_origin complement(5743..6331) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6502..7362) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" promoter complement(7363..7467) /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 ccatgagcgg gggcgaggag ctgttcgccg gcatcgtgcc cgtgctgatc 2701 gagctggacg gcgacgtgca cggccacaag ttcagcgtgc gcggcgaggg cgagggcgac 2761 gccgactacg gcaagctgga gatcaagttc atctgcacca ccggcaagct gcccgtgccc 2821 tggcccaccc tggtgaccac cctctgctac ggcatccagt gcttcgcccg ctaccccgag 2881 cacatgaaga tgaacgactt cttcaagagc gccatgcccg agggctacat ccaggagcgc 2941 accatccagt tccaggacga cggcaagtac aagacccgcg gcgaggtgaa gttcgagggc 3001 gacaccctgg tgaaccgcat cgagctgaag ggcaaggact tcaaggagga cggcaacatc 3061 ctgggccaca agctggagta cagcttcaac agccacaacg tgtacatccg ccccgacaag 3121 gccaacaacg gcctggaggc taacttcaag acccgccaca acatcgaggg cggcggcgtg 3181 cagctggccg accactacca gaccaacgtg cccctgggcg acggccccgt gctgatcccc 3241 atcaaccact acctgagcac tcagaccaag atcagcaagg accgcaacga ggcccgcgac 3301 cacatggtgc tcctggagtc cttcagcgcc tgctgccaca cccacggcat ggacgagctg 3361 tacaggggat ccgagggcag aggaagcctt ctaacatgcg gtgacgtgga ggagaatccc 3421 ggcccttccg ggatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc 3481 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc 3541 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc 3601 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg 3661 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc 3721 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac 3781 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc 3841 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg 3901 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc 3961 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag 4021 cccggtgcct gaatctaggt cgacaatcaa cctctggatt acaaaatttg tgaaagattg 4081 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct 4141 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg 4201 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact 4261 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc 4321 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc 4381 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa 4441 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc 4501 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg 4561 gctctgcggc ctcttccgcg tctccgcctt cgccctcaga cgagtcggat ctctctttgg 4621 gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg tagatcttag 4681 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagataaga 4741 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc 4801 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt 4861 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc 4921 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact 4981 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta 5041 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 5101 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc 5161 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc 5221 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag 5281 aagtagtgag gaggcttttt tggaggccta gacttttgca gagaccaaat tcgtaatcat 5341 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 5401 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 5461 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 5521 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 5581 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 5641 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 5701 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 5761 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 5821 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 5881 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 5941 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 6001 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 6061 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 6121 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 6181 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 6241 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 6301 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 6361 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 6421 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 6481 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 6541 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 6601 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 6661 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 6721 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 6781 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 6841 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 6901 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 6961 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 7021 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 7081 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 7141 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 7201 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 7261 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 7321 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 7381 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 7441 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta 7501 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg 7561 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt 7621 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg 7681 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt 7741 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg 7801 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 7861 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 7921 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg //