LOCUS Exported 7714 bp ds-DNA circular SYN 23-SEP-2022 DEFINITION lentiviral vector with Flag, HA tags. ACCESSION . VERSION . KEYWORDS pCDH-EF1-FHC SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7714) AUTHORS Yousefzadeh MJ, Wyatt DW, Takata K, Mu Y, Hensley SC, Tomida J, Bylund GO, Doublie S, Johansson E, Ramsden DA, McBride KM, Wood RD TITLE Mechanism of suppression of chromosomal instability by DNA polymerase POLQ. JOURNAL PLoS Genet. 2014 Oct 2;10(10):e1004654. doi: 10.1371/journal.pgen.1004654. eCollection 2014 Oct. PUBMED 25275444 REFERENCE 2 (bases 1 to 7714) AUTHORS . TITLE Direct Submission JOURNAL Exported Sep 23, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7714 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 144..161 /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" CDS complement(339..365) /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS complement(381..404) /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" LTR 476..744 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" promoter complement(757..968) /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" misc_feature 1011..1127 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" CDS complement(1394..1438) /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 1623..1856 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." misc_feature 2349..2474 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" LTR 2521..2701 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" promoter complement(2702..2928) /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" primer_bind complement(2943..2960) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2943..2959) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2952..2974) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(3168..3187) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3287..3309 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3347..3365) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3433..3537 /gene="bla" /label=AmpR promoter CDS 3538..4398 /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(3756..3775) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4569..5157 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5058..5077 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5311..5328 /label=L4440 /note="L4440 vector, forward primer" protein_bind 5445..5466 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5481..5511 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5519..5535 /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 5524..5546 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 5543..5559 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5543..5559 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" rep_origin 5592..5727 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(5646..5665) /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 5767..5888 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5778..5797) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 5832..5851 /label=SV40pA-R /note="SV40 polyA, reverse primer" LTR 5960..6193 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" misc_feature 6267..6855 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" CDS 6373..6384 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind 6782..6802 /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6862..7461) /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(6945..6965) /label=Puro-F /note="Puromycin resistance gene, forward primer" primer_bind 7442..7461 /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" misc_feature 7470..328 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(7629..7648) /label=IRES-F /note="IRES internal ribosome entry site, forward primer" ORIGIN 1 atcttataca cgtggctttt ggccgcagag gcacctgtcg ccaggtgggg ggttccgctg 61 cctgcaaagg gtcgctacag acgttgtttg tcttcaagaa gcttccagag gaactgcttc 121 cttcacgaca ttcaacagac cttgcattcc tttggcgaga ggggaaagac ccctaggaat 181 gctcgtcaag aagacagggc caggtttccg ggccctcaca ttgccaaaag acggcaatat 241 ggtggaaaat aacatataga caaacgcaca ccggccttat tccaagcggc ttcggccagt 301 aacgttangg gggggggagg gagaggggcg cggccctagg cgtagtcggg cacgtcgtag 361 gggtatcctc cagcggccga cttgtcatcg tcgtccttgt agtctcctcc agcggccgcg 421 gatccgattt aaattcgaat tcgctagctc tagagtaggc gccggtcaca gcttggatct 481 gtaacggcgc agaacagaaa acgaaacaaa gacgtagagt tgagcaagca gggtcaggca 541 aagcgtggag agccggctga gtctaggtag gctccaaggg agcgccggac aaaggcccgg 601 tctcgacctg agctttaaac ttacctagac ggcggacgca gttcaggagg caccacaggc 661 gggaggcggc agaacgcgac tcaaccggcg tggatggcgg cctcaggtag ggcggcgggc 721 gcgtgaagga gagatgcgag cccctcgaag cttcagctgt gttctggcgg caaacccgtt 781 gcgaaaaaga acgttcacgg cgactactgc acttatatac ggttctcccc caccctcggg 841 aaaaaggcgg agccagtaca cgacatcact ttcccagttt accccgcgcc accttctcta 901 ggcacccgtt caattgccga cccctccccc caacttctcg gggactgtgg gcgatgtgcg 961 ctctgcccac tgacgggcac cggagcgatc gcagatccac tagtatcgat aaaattttga 1021 attttgtaat ttgtttttgt aattctttag tttgtatgtc tgttgctatt atgtctacta 1081 ttctttcccc tgcactgtac cccccaatcc ccccttttct tttaaaagtt aaccgatacc 1141 gtcgagatcc gttcactaat cgaatggatc tgtctctgtc tctctctcca ccttcttctt 1201 ctattccttc gggcctgtcg ggtcccctcg gggttgggag gtgggtctga aacgataatg 1261 gtgaatatcc ctgcctaact ctattcacta tagaaagtac agcaaaaact attcttaaac 1321 ctaccaagcc tcctactatc attatgaata attttatata ccacagccaa tttgttatgt 1381 taaaccaatt ccacaaactt gcccatttat ctaattccaa taattcttgt tcattctttt 1441 cttgctggtt ttgcgattct tcaattaagg agtgtattaa gcttgtgtaa ttgttaattt 1501 ctctgtccca ctccatccag gtcgtgtgat tccaaatctg ttccagagat ttattactcc 1561 aactagcatt ccaaggcaca gcagtggtgc aaatgagttt tccagagcaa ccccaaatcc 1621 ccaggagctg ttgatccttt aggtatcttt ccacagccag gattcttgcc tggagctgct 1681 tgatgcccca gactgtgagt tgcaacagat gctgttgcgc ctcaatagcc ctcagcaaat 1741 tgttctgctg ctgcactata ccagacaata attgtctggc ctgtaccgtc agcgtcattg 1801 acgctgcgcc catagtgctt cctgctgctc ccaagaaccc aaggaacaaa gctcctattc 1861 ccactgctct tttttctctc tgcaccactc ttctctttgc cttggtgggt gctactccta 1921 atggttcaat ttttactact ttatatttat ataattcact tctccaattg tccctcatat 1981 ctcctcctcc aggtctgaag atcagtggcc gcttgctgtg cggtggtctt acttttgttt 2041 tgctcttcct ctatcttgtc taaagcttcc ttggtgtctt ttatctctat cctttgatgc 2101 acacaataga gggttgctac tgtattatat aatgatctaa gttcttctga tcctgtctga 2161 agggatggtt gtagctgtcc cagtatttgt ctacagcctt ctgatgtttc taacaggcca 2221 ggattaactg cgaatcgttc tagctccctg cttgcccata ctatatgttt taatttatat 2281 tttttctttc cccctggcct taaccgaatt ttttcccatc gcgatctaat tctcccccgc 2341 ttaatactga cgctctcgca cccatctctc tccttctagc ctccgctagt caaaattttt 2401 ggcgtactca ccagtcgccg cccctcgcct cttgccgtgc gcgcttcagc aagccgagtc 2461 ctgcgtcgag agagctcctc tggtttccct ttcgctttca agtccctgtt cgggcgccac 2521 tgctagagat tttccacact gactaaaagg gtctgaggga tctctagtta ccagagtcac 2581 acaacagacg ggcacacact acttgaagca ctcaaggcaa gctttattga ggcttaagca 2641 gtgggttccc tagttagcca gagagctccc aggctcagat ctggtctaac cagagagacc 2701 cgtttatgta tcgagctagg cacttaaata caatatctct gcaatgcggc aattcagtgg 2761 ttcgtccaat ccatgtcaga cccgtctgtt gccttcctaa taaggcacga tcgtaccacc 2821 ttacttccac caatcggcat gcacggtgct ttttctctcc ttgtaaggca tgttgctaac 2881 tcatcgttac catgttgcaa gactacaaga gtattgcata agactacacg cgtcagcttg 2941 gcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat 3001 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat 3061 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc 3121 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct 3181 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg 3241 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg 3301 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc 3361 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt 3421 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat 3481 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg 3541 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt 3601 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga 3661 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa 3721 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt 3781 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt 3841 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc 3901 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga 3961 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat 4021 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct 4081 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc 4141 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg 4201 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc 4261 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg 4321 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca 4381 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta 4441 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc 4501 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa 4561 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 4621 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 4681 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc 4741 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 4801 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 4861 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 4921 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 4981 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 5041 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 5101 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 5161 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc 5221 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat 5281 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag 5341 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac 5401 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc 5461 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt 5521 gtgagcggat aacaatttca cacaggaaac agctatgaca tgattacgaa tttggtctct 5581 gcaaaagtct aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga 5641 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc 5701 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggatag ctagagccag 5761 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat 5821 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata 5881 aacaagttcc tctcactctc tgatattcat ttctttgcaa gttataaata ctgaataata 5941 agatgacatg aactactact gctagagatt ttccacactg actaaaaggg tctgagggat 6001 ctctagttac cagagtcaca caacagacgg gcacacacta cttgaagcac tcaaggcaag 6061 ctttattgag gcttaagcag tgggttccct agttagccag agagctccca ggctcagatc 6121 tggtctaacc agagagaccc agtacaagca aaaagcagat cttatcttcg ttgggagtga 6181 attagccctt ccagtccccc cttttctttt aaaaagtggc taagatctac agctgccttg 6241 taagtcattg gtcttaaagg taccaggcgg ggaggcggcc caaagggaga tccgactcgt 6301 ctgagggcga aggcgaagac gcggaagagg ccgcagagcc ggcagcaggc cgcgggaagg 6361 aaggtccgct ggattgaggg ccgaagggac gtagcagaag gacgtcccgc gcagaatcca 6421 ggtggcaaca caggcgagca gccaaggaaa ggacgatgat ttccccgaca acaccacgga 6481 attgtcagtg cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag 6541 ttccgccgtg gcaataggga gggggaaagc gaaagtcccg gaaaggagct gacaggtggt 6601 ggcaatgccc caaccagtgg gggttgcgtc agcaaacaca gtgcacacca cgccacgttg 6661 cctgacaacg ggccacaact cctcataaag agacagcaac caggatttat acaaggagga 6721 gaaaatgaaa gccatacggg aagcaatagc atgatacaaa ggcattaaag cagcgtatcc 6781 acatagcgta aaaggagcaa catagttaag aataccagtc aatctttcac aaattttgta 6841 atccagaggt tgattgtcga cctaggcacc gggcttgcgg gtcatgcacc aggtgcgcgg 6901 tccttcgggc acctcgacgt cggcggtgac ggtgaagccg agccgctcgt agaaggggag 6961 gttgcggggc gcggaggtct ccaggaaggc gggcaccccg gcgcgctcgg ccgcctccac 7021 tccggggagc acgacggcgc tgcccagacc cttgccctgg tggtcgggcg agacgccgac 7081 ggtggccagg aaccacgcgg gctccttggg ccggtgcggc gccaggaggc cttccatctg 7141 ttgctgcgcg gccagccggg aaccgctcaa ctcggccatg cgcgggccga tctcggcgaa 7201 caccgccccc gcttcgacgc tctccggcgt ggtccagacc gccaccgcgg cgccgtcgtc 7261 cgcgacccac accttgccga tgtcgagccc gacgcgcgtg aggaagagtt cttgcagctc 7321 ggtgacccgc tcgatgtggc ggtccggatc gacggtgtgg cgcgtggcgg ggtagtcggc 7381 gaacgcggcg gcgagggtgc gtacggccct ggggacgtcg tcgcgggtgg cgaggcgcac 7441 cgtgggcttg tactcggtca ttccggaggt attatcgtgt ttttcaaagg aaaaccacgt 7501 ccccgtggtt cggggggcct agacgttttt ttaacctcga ctaaacacat gtaaagcatg 7561 tgcaccgagg ccccagatca gatcccatac aatggggtac cttctgggca tccttcagcc 7621 ccttgttgaa tacgcttgag gagagccatt tgactctttc cacaactatc caactcacaa 7681 cgtggcactg gggttgtgcc gcctttgcag gtgt //