LOCUS pTwist_Lenti_SFF 7718 bp DNA circular SYN 11-NOV-2024 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7718) AUTHORS siwong TITLE Direct Submission JOURNAL Exported Dec 5, 2024 from SnapGene 8.0.0 https://www.snapgene.com FEATURES Location/Qualifiers source 1..7718 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 1..408 /label=SFFV promoter /note="spleen focus-forming virus long terminal repeat (LTR) promoter" misc_feature 427^428 /label=Your gene here misc_feature 465..1053 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" CDS complement(936..947) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 1125..1358 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 1436..1557 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin 1597..1732 /label=SV40 ori /note="SV40 origin of replication" promoter complement(1753..1771) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1781..1797) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" rep_origin 1939..2394 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 2420..2524 /gene="bla" /label=AmpR promoter CDS join(2525..2593,2594..3385) /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" rep_origin 3556..4144 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 4432..4453 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter join(4468..4473,4474..4491,4492..4498) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4506..4522 /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 4530..4546 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" promoter 4567..4585 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 4613..4839 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 4840..5020 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 5067..5192 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 5685..5918 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 6103..6147 /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" promoter 6322..6822 /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" CDS 6844..7443 /codon_start=1 /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" CDS 6844..7443 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" misc_feature 7485..7602 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" ORIGIN 1 gtaacgccat tttgcaaggc atggaaaaat accaaaccaa gaatagagaa gttcagatca 61 agggcgggta catgaaaata gctaacgttg ggccaaacag gatatctgcg gtgagcagtt 121 tcggccccgg cccggggcca agaacagatg gtcaccgcag tttcggcccc ggcccgaggc 181 caagaacaga tggtccccag atatggccca accctcagca gtttcttaag acccatcaga 241 tgtttccagg ctcccccaag gacctgaaat gaccctgcgc cttatttgaa ttaaccaatc 301 agcctgcttc tcgcttctgt tcgcgcgctt ctgcttcccg agctctataa aagagctcac 361 aacccctcac tcggcgcgcc agtcctccga cagactgagt cgcccgggga gacccaagct 421 ggctagctct agagggcccg tttaaacccg ctgatcagcc tcgaaatcaa cctctggatt 481 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg 541 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct 601 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc 661 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca 721 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac 781 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt 841 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct 901 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc 961 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga 1021 cgagtcggat ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa 1081 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca 1141 ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat 1201 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt 1261 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc 1321 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt acgtatagta gttcatgtca 1381 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt 1441 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 1501 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 1561 tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt 1621 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc 1681 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagggacg 1741 tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac 1801 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt 1861 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca 1921 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg 1981 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct 2041 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc 2101 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg 2161 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt 2221 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg 2281 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc 2341 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg 2401 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa 2461 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa 2521 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct 2581 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg 2641 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg 2701 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt 2761 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga 2821 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga 2881 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac 2941 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg 3001 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac 3061 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct 3121 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct 3181 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg 3241 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat 3301 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg 3361 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat 3421 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct 3481 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa 3541 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa 3601 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc 3661 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta 3721 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct 3781 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg 3841 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag 3901 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc 3961 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg 4021 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt 4081 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg 4141 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca 4201 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg 4261 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc 4321 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag 4381 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag 4441 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg 4501 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa 4561 gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctgcaagctt aatgtagtct 4621 tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa 4681 ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga tcgtgcctta 4741 ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc 4801 agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg gttagaccag 4861 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc 4921 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga 4981 tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact 5041 tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc 5101 gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag 5161 gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc 5221 gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag 5281 tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag 5341 aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac 5401 ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa 5461 aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg 5521 cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga 5581 agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag 5641 gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt 5701 gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag 5761 gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag 5821 gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc 5881 ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga 5941 aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa 6001 cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc 6061 ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta 6121 ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg 6181 tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct 6241 gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac 6301 ctcccaaccc cgaggggacc cttgcgcctt ttccaaggca gccctgggtt tgcgcaggga 6361 cgcggctgct ctgggcgtgg ttccgggaaa cgcagcggcg ccgaccctgg gtctcgcaca 6421 ttcttcacgt ccgttcgcag cgtcacccgg atcttcgccg ctacccttgt gggccccccg 6481 gcgacgcttc ctgctccgcc cctaagtcgg gaaggttcct tgcggttcgc ggcgtgccgg 6541 acgtgacaaa cggaagccgc acgtctcact agtaccctcg cagacggaca gcgccaggga 6601 gcaatggcag cgcgccgacc gcgatgggct gtggccaata gcggctgctc agcagggcgc 6661 gccgagagca gcggccggga aggggcggtg cgggaggcgg ggtgtggggc ggtagtgtgg 6721 gccctgttcc tgcccgcgcg gtgttccgca ttctgcaagc ctccggagcg cacgtcggca 6781 gtcggctccc tcgttgaccg aatcaccgac ctctctcccc agggggtacc accggagctt 6841 accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc 6901 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg 6961 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc 7021 gacatcggca aggtgtgggt cgcggacgac ggcgccgccg tggcggtctg gaccacgccg 7081 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc 7141 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag 7201 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg 7261 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc 7321 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc 7381 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc 7441 tgagaattct agatcttgag acaaatggca gtattcatcc acaattttaa aagaaaaggg 7501 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa 7561 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac 7621 agcagagatc cactttggcg ccggctcgag ggggcccggg gttattaata gtaatcaatt 7681 acggggtcat tagttcatag cccatatatg gagttccg //