LOCUS Exported 8432 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expressed barcode for sample multiplexing in single-cell analysis. ACCESSION . VERSION . KEYWORDS CellTagA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8432) TITLE CellTag Indexing: genetic barcode-based sample multiplexing for single-cell genomics JOURNAL Unpublished REFERENCE 2 (bases 1 to 8432) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8432 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 314..517 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 468..488 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 532..712 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 759..884 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1377..1610 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1795..1839 /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 2106..2223 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" polyA_signal 2355..2489 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" CDS complement(2543..3259) /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2585..2606) /label=EGFP-C /note="EGFP, forward primer" primer_bind 2934..2953 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3193..3214 /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3342..3366) /label=LNCX /note="Human CMV promoter, forward primer" promoter complement(3363..3401) /label=minimal CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" protein_bind 3835..3868 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind 3883..4007 /gene="mutant version of attR" /label=attR1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" promoter 4032..4062 /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" primer_bind complement(4080..4096) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4080..4096) /label=pBluescriptSK /note="For pBluescript vector" protein_bind 4097..4130 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 4136..4724 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4189..4209) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4607..4618) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 4811..5044 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 5116..5237 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind 5207..5226 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 5277..5412 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5339..5358 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(5432..5451) /label=T7 /note="T7 promoter, forward primer" promoter complement(5433..5451) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5461..5478) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(5461..5477) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5470..5492) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 5619..6074 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5706..5725) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5916..5937 /label=F1ori-F /note="F1 origin, forward primer" promoter 6100..6204 /gene="bla" /label=AmpR promoter CDS 6205..7065 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(6423..6442) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7236..7824 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7725..7744 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7978..7995 /label=L4440 /note="L4440 vector, forward primer" protein_bind 8112..8133 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 8148..8178 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8186..8202 /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 8191..8213 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 8210..8226 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 8210..8226 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 8245..8265 /label=T3 /note="T3 promoter, forward primer" promoter 8247..8265 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" enhancer 8366..313 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" ORIGIN 1 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc 61 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca 121 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta 181 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta 241 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat 301 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga 361 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca 421 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg 481 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc ggggtctctc 541 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag 601 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct 661 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc 721 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc 781 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt 841 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga 901 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat 961 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt 1021 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag 1081 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa 1141 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa 1201 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg 1261 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta 1321 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga 1381 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg 1441 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg 1501 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc 1561 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg 1621 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat 1681 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac 1741 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat 1801 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca 1861 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga 1921 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg 1981 tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa 2041 ggtggagaga gagacagaga cagatccatt cgattagtga acggatctcg acggtatcgg 2101 ttaactttta aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac 2161 ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat 2221 tttatcgatc acgagactag cctcgagtga tcataatcag ccataccaca tttgtagagg 2281 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg 2341 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa ggcaatagca 2401 tcacaaattt cacaaataag gcattttttt cactgcattc tagttttggt ttgtccaaac 2461 tcatcaatgt atcttatcat gtctggatct caaatccctc ggaagctgcg cctgtcatcg 2521 aattcatata gcaaccggtt tacttgtaca gctcgtccat gccgagagtg atcccggcgg 2581 cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcaggg 2641 cggactgggt gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg 2701 tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg tggcggatct 2761 tgaagttcac cttgatgccg ttcttctgct tgtcggccat gatatagacg ttgtggctgt 2821 tgtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct 2881 tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt 2941 agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg 3001 acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt 3061 aggtcagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatga 3121 acttcagggt cagcttaccg taggtggcat cgccctcgcc ctcgccggac acgctgaact 3181 tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct 3241 cctcgccctt gctcaccatg gtggtaccga attccgcgga ggctggatcg gtcccggtgt 3301 cttctatgga ggtcaaaaca gcgtggatgg cgtctccagg cgatctgacg gttcactaaa 3361 cgagctctgc ttatataggc ctcccaccgt acacgcctac cactagtaat gaaagacccc 3421 acctgtaggt ttggcaagct agctgcagta acgccatttt gcaaggcatg gaaaaatacc 3481 aaaccaagaa tagagaagtt cagatcaagg gcgggtacat gaaaatagct aacgttgggc 3541 caaacaggat atctgcggtg agcagtttcg gccccggccc ggggccaaga acagatggtc 3601 accgcagttt cggccccggc ccgaggccaa gaacagatgg tccccagata tggcccaacc 3661 ctcagcagtt tcttaagacc catcagatgt ttccaggctc ccccaaggac ctgaaatgac 3721 cctgcgcctt atttgaatta accaatcagc ctgcttctcg cttctgttcg cgcgcttctg 3781 cttcccgagc tctataaaag agctcacaac ccctcactcg gcttaattaa gatcataact 3841 tcgtatagca tacattatac gaagttatcc cagcttgata tcacaagttt gtacaaaaaa 3901 gctgaacgag aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa 3961 aaacagacta cataatactg taaaacacaa catatccagt cactatggcg gccgcattag 4021 gcaccccagg ctttacactt tatgcttccg gctcgtataa tgtgtggatt ttgagttagg 4081 atccactagt tctagaataa cttcgtatag catacattat acgaagttat tcgacaatca 4141 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt 4201 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc 4261 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc 4321 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg 4381 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc 4441 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg 4501 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg 4561 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc 4621 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct 4681 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttcgagctcg 4741 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa 4801 aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt 4861 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac 4921 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg 4981 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct 5041 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata 5101 tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc 5161 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa 5221 ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccatcc 5281 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta 5341 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct 5401 tttttggagg cctagggacg tacccaattc gccctatagt gagtcgtatt acgcgcgctc 5461 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg 5521 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg 5581 cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt 5641 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc 5701 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca 5761 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc 5821 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt 5881 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac 5941 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc 6001 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt 6061 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta 6121 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt 6181 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc 6241 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa 6301 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt 6361 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt 6421 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc 6481 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg 6541 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg 6601 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac 6661 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca 6721 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta 6781 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat 6841 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa 6901 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag 6961 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat 7021 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt 7081 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg 7141 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga 7201 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta 7261 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa 7321 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact 7381 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca 7441 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt 7501 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg 7561 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag 7621 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta 7681 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat 7741 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg 7801 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc 7861 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac 7921 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc 7981 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt 8041 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag 8101 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg 8161 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc 8221 tatgaccatg attacgccaa gcgcgcaatt aaccctcact aaagggaaca aaagctggag 8281 ctgcaagctt ggccattgca tacgttgtat ccatatcata atatgtacat ttatattggc 8341 tcatgtccaa cattaccgcc atgttgacat tgattattga ctagttatta atagtaatca 8401 attacggggt cattagttca tagcccatat at //