LOCUS Exported 6966 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Dox-inducible lentivral vector expressing mCherry red fluorescent protein. ACCESSION . VERSION . KEYWORDS pHAGE2-TetOminiCMV-mCherry SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6966) AUTHORS Velychko S, Adachi K, Kim KP, Hou Y, MacCarthy CM, Wu G, Scholer HR TITLE Excluding Oct4 from Yamanaka Cocktail Unleashes the Developmental Potential of iPSCs. JOURNAL Cell Stem Cell. 2019 Oct 30. pii: S1934-5909(19)30423-0. doi: 10.1016/j.stem.2019.10.002. PUBMED 31708402 REFERENCE 2 (bases 1 to 6966) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6966 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 2..272 /label=tetracycline response element /bound_moiety="tetracycline repressor TetR" /note="contains seven copies of the tetracycline operator tetO" primer_bind 340..364 /label=LNCX /note="Human CMV promoter, forward primer" CDS 435..1145 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(580..598) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 848..867 /label=mCherry-F /note="mCherry, forward primer" misc_feature 1159..1747 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(1212..1232) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(1630..1641) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 1895..2075 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(2122..2140) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 2208..2312 /gene="bla" /label=AmpR promoter CDS 2313..3173 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(2531..2550) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3344..3932 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3833..3852 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4086..4103 /label=L4440 /note="L4440 vector, forward primer" primer_bind 4195..4214 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" protein_bind 4343..4364 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4379..4409 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4417..4433 /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 4422..4444 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 4441..4457 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 4441..4457 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 4507..4526 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" LTR 4718..5351 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 5398..5523 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 6020..6253 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 6438..6482 /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" misc_feature 6775..6892 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" ORIGIN 1 ctccctatca gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga 61 gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt 121 ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt 181 gatagagaaa agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag 241 tcgagtttac cactccctat cagtgataga gaaaagtgaa agtcgagctc ggtacccggg 301 tcgaggtagg catgtacggt gggaggccta tataagcaga gctcgtttag tgaaccgtca 361 gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc 421 cagcctgcgg ccgcatggtg agcaagggcg aggaggataa catggccatc atcaaggagt 481 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg 541 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg 601 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg 661 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct 721 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact 781 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct 841 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt 901 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg 961 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg 1021 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg 1081 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca 1141 agtaaatcga tagatcctaa tcaacctctg gattacaaaa tttgtgaaag attgactggt 1201 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat 1261 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg 1321 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt 1381 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact 1441 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc 1501 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg 1561 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc 1621 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg 1681 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc 1741 tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt 1801 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatcacct 1861 gcaggacagg cgcgccctgc tttttgcttg tactgggtct ctctggttag accagatctg 1921 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc 1981 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct 2041 cagacccttt tagtcagtgt ggaaaatctc tagcacccgg gcgattaagg aaagggctag 2101 atcattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga 2161 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta 2221 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat 2281 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc 2341 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga 2401 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca 2461 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt 2521 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg 2581 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc 2641 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata 2701 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt 2761 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag 2821 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca 2881 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg 2941 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg 3001 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag 3061 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg 3121 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag 3181 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga 3241 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt 3301 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc 3361 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc 3421 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac 3481 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac 3541 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt 3601 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct 3661 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat 3721 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt 3781 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg 3841 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt 3901 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt 3961 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg 4021 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg 4081 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc 4141 ccgcgcgttg gccgattcat taatgcagca agctcatggc tgactaattt tttttattta 4201 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt 4261 tggaggccta ggcttttgca aaaagctccc cgtggcacga caggtttccc gactggaaag 4321 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt 4381 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca 4441 caggaaacag ctatgacatg attacgaatt tcacaaataa agcatttttt tcactgcatt 4501 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc aactggataa 4561 ctcaagctaa ccaaaatcat cccaaacttc ccaccccata ccctattacc actgccaatt 4621 acctgtggtt tcatttactc taaacctgtg attcctctga attattttca ttttaaagaa 4681 attgtatttg ttaaatatgt actacaaact tagtagttgg aagggctaat tcactcccaa 4741 agaagacaag atatccttga tctgtggatc taccacacac aaggctactt ccctgattag 4801 cagaactaca caccagggcc aggggtcaga tatccactga cctttggatg gtgctacaag 4861 ctagtaccag ttgagccaga taaggtagaa gaggccaata aaggagagaa caccagcttg 4921 ttacaccctg tgagcctgca tgggatggat gacccggaga gagaagtgtt agagtggagg 4981 tttgacagcc gcctagcatt tcatcacgtg gcccgagagc tgcatccgga gtacttcaag 5041 aactgctgat atcgagcttg ctacaaggga ctttccgctg gggactttcc agggaggcgt 5101 ggcctgggcg ggactgggga gtggcgagcc ctcagatcct gcatataagc agctgctttt 5161 tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact 5221 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc 5281 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa 5341 aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc 5401 tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg 5461 gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc 5521 gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg 5581 ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc 5641 gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta 5701 caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc 5761 ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata 5821 gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccggccg ctgatcttca 5881 gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag 5941 taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag 6001 aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca 6061 ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag 6121 tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca 6181 cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg 6241 atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc 6301 cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga 6361 tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat 6421 cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt 6481 tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag 6541 taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta 6601 ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca 6661 ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag 6721 tgaacggatc tcgacggtat cgccgaattc acaaatggca gtattcatcc acaattttaa 6781 aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac 6841 agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta 6901 ttacagggac agcagagatc cagtttggac tagtccacac cctaactgac acactcgagt 6961 ttacca //