LOCUS Cellecta-pRSITH1 8846 bp ds-DNA circular SYN 17-MAY-2019 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8846) AUTHORS Cellecta TITLE Direct Submission JOURNAL Exported Friday, May 17, 2019 from SnapGene 4.3.7 to Vector NTI(R) format https://www.snapgene.com COMMENT VNTDATE|309830400| COMMENT VNTDBDATE|877910400| COMMENT VNTNAME|Cellecta-pRSITH1PGP-H1Tet-sh-HTS3-PGK-TetRep-2A-TagGFP2-2A-P uro.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 1803..2021 /label=H1-Tet /note="doxycycline-inducible variant of the human H1 RNA promoter (Henriksen et al., 2007)" protein_bind 1975..1993 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2003..2021 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" misc_feature 2022..2047 /label=cloning site misc_feature 2151..2268 /label=cPPT /note="central polypurine tract and central termination sequence of HIV-1" promoter 2321..2821 /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" CDS 2834..3454 /codon_start=1 /gene="synthetic gene" /product="tetracycline repressor TetR" /label=TetR /note="TetR binds to the tetracycline operator tetO to inhibit transcription. This inhibition can be relieved by adding tetracycline or doxycycline." CDS 3461..3514 /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 3521..4234 /codon_start=1 /product="monomeric green fluorescent protein, also known as mTagGFP" /label=TagGFP2 /note="mammalian codon-optimized" CDS 4253..4306 /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 4313..4912 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" misc_feature 4925..5513 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" LTR 5587..5820 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 5892..6013 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin 6053..6188 /label=SV40 ori /note="SV40 origin of replication" rep_origin complement(6623..7211) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(7382..8242) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" promoter complement(8243..8347) /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 atgaacgctg acgtcatcaa cccgctccaa ggaatcgcgg gcccagtgtc actaggcggg 1861 aacacccagc gcgcgtgcgc cctggcagga agatggctgt gagggacagg ggagtggcgc 1921 cctgcaatat ttgcatgtcg ctatgtgttc tgggaaatca ccataaacgt gaaatcccta 1981 tcagtgatag agacttataa gttccctatc agtgatagag aaccggagtc ttcttttttg 2041 aagacacttc gtgcgtttgg ggtttcggac tgtagaactc tgaacctctc ggtggtcgcc 2101 gtatcattag aattctcgac ctcgagacaa atggcagtat tcatccacaa ttttaaaaga 2161 aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac 2221 atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac 2281 agggacagca gagatccact ttggccgccg ttcgaagggg ttgcgccttt tccaaggcag 2341 ccctgggttt gcgcagggac gcggctgctc tgggcgtggt tccgggaaac gcagcggcgc 2401 cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc gtcacccgga tcttcgccgc 2461 tacccttgtg ggccccccgg cgacgcttcc tgctccgccc ctaagtcggg aaggttcctt 2521 gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca cgtctcacta gtaccctcgc 2581 agacggacag cgccagggag caatggcagc gcgccgaccg cgatgggctg tggccaatag 2641 cggctgctca gcagggcgcg ccgagagcag cggccgggaa ggggcggtgc gggaggcggg 2701 gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg tgttccgcat tctgcaagcc 2761 tccggagcgc acgtcggcag tcggctccct cgttgaccga atcaccgacc tctctcccca 2821 gggtctagac accatgtcca gactggacaa gagcaaagtc atcaactctg ccttggagct 2881 cctgaatgaa gttggcattg agggcttgac caccaggaag ctggcccaga agctgggtgt 2941 ggagcagcct accctgtact ggcatgtgaa gaacaagagg gctctgcttg atgccctggc 3001 cattgagatg ttggacaggc accacaccca cttctgccct ctggaagggg agtcctggca 3061 ggacttcctg aggaacaatg ccaagagctt cagatgtgcc ttgctctccc accgggacgg 3121 tgccaaagtt cacttgggca ccaggcctac agagaagcag tatgagaccc tggagaacca 3181 gctggcattc ctgtgccaac aaggcttctc cctggaaaat gccttgtatg ccctctctgc 3241 tgtgggccac ttcaccttgg gctgtgtgct ggaggaccag gagcaccaag ttgccaagga 3301 ggagagggag acccccacca ctgactccat gccaccactg ctgcggcaag ctattgagtt 3361 gtttgaccac caaggggctg agcctgcatt cctgtttggc ctggaactga tcatctgtgg 3421 cctggaaaag cagctgaaat gtgagtctgg ctctggatcc gagggcagag gaagccttct 3481 aacatgcggt gacgtggagg agaatcccgg cccttccgga atgagcgggg gcgaggagct 3541 gttcgccggc atcgtgcccg tgctgatcga gctggacggc gacgtgcacg gccacaagtt 3601 cagcgtgcgc ggcgagggcg agggcgacgc cgactacggc aagctggaga tcaagttcat 3661 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctg gtgaccaccc tctgctacgg 3721 catccagtgc ttcgcccgct accccgagca catgaagatg aacgacttct tcaagagcgc 3781 catgcccgag ggctacatcc aggagcgcac catccagttc caggacgacg gcaagtacaa 3841 gacccgcggc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg 3901 caaggacttc aaggaggacg gcaacatcct gggccacaag ctggagtaca gcttcaacag 3961 ccacaacgtg tacatccgcc ccgacaaggc caacaacggc ctggaggcta acttcaagac 4021 ccgccacaac atcgagggcg gcggcgtgca gctggccgac cactaccaga ccaacgtgcc 4081 cctgggcgac ggccccgtgc tgatccccat caaccactac ctgagcactc agaccaagat 4141 cagcaaggac cgcaacgagg cccgcgacca catggtgctc ctggagtcct tcagcgcctg 4201 ctgccacacc cacggcatgg acgagctgta cagggctagc accggtagat ccgagggcag 4261 aggaagcctt ctaacatgcg gtgacgtgga ggagaatccc ggcccttccg ggatgaccga 4321 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct 4381 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat 4441 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa 4501 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga 4561 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct 4621 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg 4681 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt 4741 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc 4801 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga 4861 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaatctaggt 4921 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt 4981 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc 5041 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga 5101 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc 5161 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct 5221 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg 5281 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct 5341 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc 5401 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg 5461 tctccgcctt cgccctcaga cgagtcggat ctctctttgg gccgcctccc cgcctggtac 5521 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg 5581 ggggactgga agggctaatt cactcccaac gaagataaga tctgcttttt gcttgtactg 5641 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac 5701 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt 5761 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca 5821 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag 5881 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 5941 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 6001 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc 6061 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta 6121 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt 6181 tggaggccta gacttttgca gagaccaaat tcgtaatcat gtcatagctg tttcctgtgt 6241 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag 6301 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt 6361 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag 6421 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 6481 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 6541 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 6601 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 6661 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 6721 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 6781 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 6841 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 6901 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 6961 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 7021 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 7081 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 7141 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa 7201 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa 7261 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt 7321 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca 7381 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca 7441 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc 7501 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa 7561 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc 7621 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca 7681 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat 7741 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag 7801 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac 7861 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt 7921 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt 7981 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc 8041 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat 8101 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca 8161 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga 8221 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg 8281 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg 8341 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga 8401 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg 8461 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg 8521 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct 8581 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa 8641 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc 8701 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag 8761 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt 8821 gtaaaacgac ggccagtgcc aagctg //