LOCUS Exported 9334 bp ds-DNA circular SYN 06-JAN-2022 DEFINITION Expression of human FOXA1 with C-terminal V5 tag, puromycin selection marker, 3rd generation lentiviral vector. ACCESSION . VERSION . KEYWORDS pLX302_FOXA1-V5 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9334) TITLE AR Cistrome Nature Genetics JOURNAL Unpublished REFERENCE 2 (bases 1 to 9334) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 6, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9334 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 133..154 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 169..199 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 207..223 /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 212..234 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 231..247 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 231..247 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 266..286 /label=T3 /note="T3 promoter, forward primer" promoter 268..286 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 314..540 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 541..721 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 768..893 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1386..1619 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1804..1848 /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 2023..2523 /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" primer_bind 2442..2460 /label=hPGK-F /note="Human PGK promoter, forward primer" CDS 2545..3144 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(2545..2564) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 3041..3061 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 3186..3303 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" enhancer 3420..3723 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 3724..3927 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 3878..3898 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 3950..3974 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(5391..5415) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 5417..5458 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" misc_feature 5500..6088 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5553..5573) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5971..5982) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 6160..6393 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 6471..6592 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6508..6527) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6562..6581 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 6632..6767 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6694..6713 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(6787..6806) /label=T7 /note="T7 promoter, forward primer" promoter complement(6788..6806) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(6816..6833) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(6816..6832) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6825..6847) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 6974..7429 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7061..7080) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7271..7292 /label=F1ori-F /note="F1 origin, forward primer" promoter 7455..7559 /gene="bla" /label=AmpR promoter CDS 7560..8420 /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(7778..7797) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 8591..9179 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 9080..9099 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 9333..16 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg 61 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga 121 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat 181 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag 241 ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctgga 301 gctgcaagct taatgtagtc ttatgcaata ctcttgtagt cttgcaacat ggtaacgatg 361 agttagcaac atgccttaca aggagagaaa aagcaccgtg catgccgatt ggtggaagta 421 aggtggtacg atcgtgcctt attaggaagg caacagacgg gtctgacatg gattggacga 481 accactgaat tgccgcattg cagagatatt gtatttaagt gcctagctcg atacataaac 541 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca 601 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg 661 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc 721 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca 781 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc 841 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta 901 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa 961 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc 1021 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc 1081 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg 1141 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc 1201 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga 1261 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca 1321 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg 1381 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg 1441 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac 1501 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc 1561 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg 1621 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt 1681 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga 1741 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa 1801 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt 1861 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta 1921 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca 1981 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccttgcgcct tttccaaggc 2041 agccctgggt ttgcgcaggg acgcggctgc tctgggcgtg gttccgggaa acgcagcggc 2101 gccgaccctg ggtctcgcac attcttcacg tccgttcgca gcgtcacccg gatcttcgcc 2161 gctacccttg tgggcccccc ggcgacgctt cctgctccgc ccctaagtcg ggaaggttcc 2221 ttgcggttcg cggcgtgccg gacgtgacaa acggaagccg cacgtctcac tagtaccctc 2281 gcagacggac agcgccaggg agcaatggca gcgcgccgac cgcgatgggc tgtggccaat 2341 agcggctgct cagcagggcg cgccgagagc agcggccggg aaggggcggt gcgggaggcg 2401 gggtgtgggg cggtagtgtg ggccctgttc ctgcccgcgc ggtgttccgc attctgcaag 2461 cctccggagc gcacgtcggc agtcggctcc ctcgttgacc gaatcaccga cctctctccc 2521 cagggggtac caccggagct taccatgacc gagtacaagc ccacggtgcg cctcgccacc 2581 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc 2641 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc 2701 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcc 2761 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc 2821 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc 2881 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc 2941 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag 3001 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag 3061 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc 3121 atgacccgca agcccggtgc ctgagaattc tagatcttga gacaaatggc agtattcatc 3181 cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac 3241 ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat 3301 tttcgggttt attacaggga cagcagagat ccactttggc gccggctcga gggggcccgg 3361 ggttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc 3421 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg 3481 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa 3541 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca 3601 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac 3661 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc 3721 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga 3781 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg 3841 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta 3901 cggtgggagg tctatataag cagagctctc tggctaactg tcgggatcaa caagtttgta 3961 caaaaaagtt ggcatgttag gaactgtgaa gatggaaggg catgaaacca gcgactggaa 4021 cagctactac gcagacacgc aggaggccta ctcctccgtc ccggtcagca acatgaactc 4081 aggcctgggc tccatgaact ccatgaacac ctacatgacc atgaacacca tgactacgag 4141 cggcaacatg accccggcgt ccttcaacat gtcctatgcc aacccgggcc taggggccgg 4201 cctgagtccc ggcgcagtag ccggcatgcc ggggggctcg gcgggcgcca tgaacagcat 4261 gactgcggcc ggcgtgacgg ccatgggtac ggcgctgagc ccgagcggca tgggcgccat 4321 gggtgcgcag caggcggcct ccatgaatgg cctgggcccc tacgcggccg ccatgaaccc 4381 gtgcatgagc cccatggcgt acgcgccgtc caacctgggc cgcagccgcg cgggcggcgg 4441 cggcgacgcc aagacgttca agcgcagcta cccgcacgcc aagccgccct actcgtacat 4501 ctcgctcatc accatggcca tccagcaggc gcccagcaag atgctcacgc tgagcgagat 4561 ctaccagtgg atcatggacc tcttccccta ttaccggcag aaccagcagc gctggcagaa 4621 ctccatccgc cactcgctgt ccttcaatga ctgcttcgtc aaggtggcac gctccccgga 4681 caagccgggc aagggctcct actggacgct gcacccggac tccggcaaca tgttcgagaa 4741 cggctgctac ttgcgccgcc agaagcgctt caagtgcgag aagcagccgg gggccggcgg 4801 cgggggcggg agcggaagcg ggggcagcgg cgccaagggc ggccctgaga gccgcaagga 4861 cccctctggc gcctctaacc ccagcgccga ctcgcccctc catcggggtg tgcacgggaa 4921 gaccggccag ctagagggcg cgccggcccc cgggcccgcc gccagccccc agactctgga 4981 ccacagtggg gcgacggcga cagggggcgc ctcggagttg aagactccag cctcctcaac 5041 tgcgcccccc ataagctccg ggcccggggc gctggcctct gtgcccgcct ctcacccggc 5101 acacggcttg gcaccccacg agtcccagct gcacctgaaa ggggaccccc actactcctt 5161 caaccacccg ttctccatca acaacctcat gtcctcctcg gagcagcagc ataagctgga 5221 cttcaaggca tacgaacagg cactgcaata ctcgccttac ggctctacgt tgcccgccag 5281 cctgcctcta ggcagcgcct cggtgaccac caggagcccc atcgagccct cagccctgga 5341 gccggcgtac taccaaggtg tgtattccag acccgtccta aacacttcct acccaacttt 5401 cttgtacaaa gtggttggta agcctatccc taaccctctc ctcggtctcg attctacgta 5461 gtaatgagct agcgctaacc ggtggcgcgt taagtcgaca atcaacctct ggattacaaa 5521 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac 5581 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc 5641 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt 5701 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc 5761 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc 5821 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg 5881 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt 5941 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc 6001 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt 6061 cggatctccc tttgggccgc ctccccgcgt cgactttaag accaatgact tacaaggcag 6121 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc 6181 aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag 6241 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt 6301 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca 6361 gaccctttta gtcagtgtgg aaaatctcta gcagtacgta tagtagttca tgtcatctta 6421 ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta 6481 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 6541 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct 6601 ggctctagct atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc 6661 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc 6721 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc 6781 aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt 6841 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc 6901 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg 6961 aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 7021 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 7081 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta 7141 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 7201 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 7261 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat 7321 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt 7381 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt 7441 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt 7501 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta 7561 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg 7621 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac 7681 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg 7741 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc 7801 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg 7861 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat 7921 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg 7981 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg 8041 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc 8101 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt 8161 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct 8221 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc 8281 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca 8341 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct 8401 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt 8461 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga 8521 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca 8581 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac 8641 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg 8701 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag 8761 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac 8821 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt 8881 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg 8941 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc 9001 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc 9061 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc 9121 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa 9181 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt 9241 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg 9301 ataccgctcg ccgcagccga acgaccgagc gcag //