LOCUS Exported 10342 bp ds-DNA circular SYN 17-MAR-2021 DEFINITION synthetic circular DNA ACCESSION pVec1 VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10342) AUTHORS . TITLE Direct Submission JOURNAL Exported Wednesday, Apr 28, 2021 from SnapGene Viewer 5.0.8 https://www.snapgene.com FEATURES Location/Qualifiers source 1..10342 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 15..514 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(50..73) /label=pMSCv_Rv CDS 535..1134 /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 1212..1763 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" CDS 1775..2494 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind 2428..2449 /label=EGFP-C LTR 2596..3110 /label=3' LTR /note="3' long terminal repeat from murine embryonic stem cell virus" primer_bind complement(3279..3295) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 3303..3319 /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)." promoter complement(3327..3357) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3372..3393 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(3681..4269) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4440..5300) /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" promoter complement(5301..5405) /gene="bla" /label=AmpR promoter LTR 6233..6749 /label=5' LTR /note="5' long terminal repeat from murine embryonic stem cell virus" misc_feature 6813..7154 /label=MESV Psi /note="packaging signal of murine embryonic stem cell virus" CDS 7221..7637 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 7567..7589 /label=pMSCV_Fw CDS 7675..7764 /codon_start=1 /product="three tandem HA epitope tags" /label=3xHA /translation="YPYDVPDYAGYPYDVPDYAASYPYDVPDYA" misc_feature 7771..10341 /label=Ago1 primer_bind complement(8150..8168) /label=mAGO1 primer_bind 9134..9154 /label=AGO1_Fw primer_bind complement(9189..9208) /label=AGO1_Rv misc_feature 9645 /label=Mismatch (C->A) ORIGIN 1 gatccaattc taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt 61 agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac 121 atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac 181 tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca 241 aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa 301 gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca 361 gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg 421 cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc 481 gctgttctcc tcttcctcat ctccgggcct ttcgacctgc agcccaagct taccatgacc 541 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc 601 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac 661 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc 721 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc 781 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg 841 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg 901 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc 961 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc 1021 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc 1081 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacgcccg 1141 ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc caattccgcc 1201 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta 1261 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc 1321 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct 1381 gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt 1441 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa 1501 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg 1561 gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga 1621 tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac 1681 atgtgtttag tcgaggttaa aaaacgtcta ggccccccga accacgggga cgtggttttc 1741 ctttgaaaaa cacgatgata atatggccac aaccatggtg agcaagggcg aggagctgtt 1801 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag 1861 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg 1921 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt 1981 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat 2041 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac 2101 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat 2161 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca 2221 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg 2281 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat 2341 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag 2401 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg 2461 gatcactctc ggcatggacg agctgtacaa gtaatgaatt aattaagaat tatcaagctt 2521 atcgataccg tcgacctgca gccaagctta tcgataaaat aaaagatttt atttagtctc 2581 cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg 2641 ccattttgca aggcatggaa aatacataac tgagaataga gaagttcaga tcaaggttag 2701 gaacagagag acagcagaat atgggccaaa caggatatct gtggtaagca gttcctgccc 2761 cggctcaggg ccaagaacag atggtcccca gatgcggtcc cgccctcagc agtttctaga 2821 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga 2881 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata 2941 aaagagccca caacccctca ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt 3001 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt 3061 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcat gggtaacagt 3121 ttcttgaagt tggagaacaa cattctgagg gtaggagtcg aatattaagt aatcctgact 3181 caattagcca ctgttttgaa tccacatact ccaatactcc tgaaatagtt cattatggac 3241 agcgcagaag agctggggag aattaattcg taatcatggt catagctgtt tcctgtgtga 3301 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc 3361 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc 3421 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc 3481 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 3541 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 3601 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 3661 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 3721 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 3781 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 3841 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt 3901 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 3961 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 4021 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 4081 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc 4141 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 4201 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 4261 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 4321 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 4381 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 4441 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 4501 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 4561 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac 4621 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag 4681 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac 4741 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 4801 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 4861 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 4921 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 4981 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 5041 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 5101 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 5161 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 5221 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 5281 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 5341 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt 5401 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca 5461 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac 5521 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat 5581 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg 5641 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata 5701 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc 5761 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg 5821 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt 5881 aaaacgacgg cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca 5941 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc 6001 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg 6061 tgatgccggc cacgatgcgt ccggcgtaga ggcgattagt ccaatttgtt aaagacagga 6121 tatcagtggt ccaggctcta gttttgactc aacaatatca ccagctgaag cctatagagt 6181 acgagccata gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag 6241 accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa 6301 tacataactg agaatagaga agttcagatc aaggttagga acagagagac agcagaatat 6361 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat 6421 ggtccccaga tgcggtcccg ccctcagcag tttctagaga accatcagat gtttccaggg 6481 tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct 6541 cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact 6601 cggcgcgcca gtcctccgat agactgcgtc gcccgggtac ccgtattccc aataaagcct 6661 cttgctgttt gcatccgaat cgtggactcg ctgatccttg ggagggtctc ctcagattga 6721 ttgactgccc acctcggggg tctttcattt ggaggttcca ccgagatttg gagacccctg 6781 cccagggacc accgaccccc ccgccgggag gtaagctggc cagcggtcgt ttcgtgtctg 6841 tctctgtctt tgtgcgtgtt tgtgccggca tctaatgttt gcgcctgcgt ctgtactagt 6901 tagctaacta gctctgtatc tggcggaccc gtggtggaac tgacgagttc tgaacacccg 6961 gccgcaaccc tgggagacgt cccagggact ttgggggccg tttttgtggc ccgacctgag 7021 gaagggagtc gatgtggaat ccgaccccgt caggatatgt ggttctggta ggagacgaga 7081 acctaaaaca gttcccgcct ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg 7141 cgtcttgtct gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc 7201 tgtatttgtc tgaaaattag ggccagactg ttaccactcc cttaagtttg accttaggtc 7261 actggaaaga tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt 7321 gggttacctt ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca 7381 cctttaaccg agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg 7441 gacacccaga ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc 7501 cctgggtcaa gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt 7561 ctctccccct tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca 7621 ctccttctct aggcgccgga attagatctc tcgaggttaa cgaattccac catgtaccca 7681 tacgatgttc ctgactatgc gggctatccc tatgacgtcc cggactatgc agcatcctat 7741 ccatatgacg ttccagatta cgctggatcc gaagcgggac cctcgggagc agcggcaggt 7801 gcctacctgc ctcccctgca gcaggtgttt caggcacccc gccggcctgg cattggcact 7861 gtgggcaaac caatcaagct tctggccaat tactttgagg tggacattcc taagattgac 7921 gtttaccatt acgaggtgga catcaagccg gataagtgtc ctcgcagagt caaccgggag 7981 gtggtggaat acatggtcca gcatttcaaa ccgcagatct ttggggatcg caagcctgtg 8041 tatgatggaa agaagaatat ttacacggtc acagcactgc ccattggcaa tgagagggtt 8101 gactttgagg tgacaatccc cggggagggg aaggatagaa tttttaaggt ctccatcaag 8161 tggctagcca ttgtgagctg gcgcatgctg catgaagcct tggtcagtgg ccagatccct 8221 gtgcccctgg agtctgtgca agccctggat gtggccatga ggcacctggc atctatgagg 8281 tacacccctg tgggccgctc cttcttctca ccgcctgagg gctactacca cccgctgggg 8341 ggtgggcgcg aggtctggtt cggctttcac cagtctgtgc gccctgccat gtggaagatg 8401 atgctcaaca ttgatgtctc agccactgcc ttctacaaag cacagccagt gattgagttc 8461 atgtgtgagg tcctggacat caggaacata gacgaacaac ccaagcccct cacggattcc 8521 cagcgtgttc ggtttaccaa ggagataaaa ggcctgaagg tggaagtgac ccactgtgga 8581 cagatgaaga ggaaataccg cgtgtgtaat gttacccgcc gccctgctag ccatcagacg 8641 ttccccttgc agctggagag tggacagact gtggagtgca ctgtggcaca gtatttcaag 8701 cagaaatata accttcagct caagtatcct cacctgccct gcctacaagt tgggcaagaa 8761 cagaagcata cctatttgcc cctcgaggtc tgtaacattg tggctgggca gcggtgcatt 8821 aagaagctga ctgacaacca gacttcaacc atgataaagg ctacagctag gtcggcccca 8881 gacagacagg aggagatcag ccgcctgatg aagaatgcta gctacaacct ggatccctac 8941 atccaggaat ttggaatcaa agtgaaggat gacatgacgg aggtgacagg acgtgtgctg 9001 ccggcgccca tcttgcagta tggcggccgg aaccgggcca ttgctacgcc caaccaaggt 9061 gtctgggaca tgcgcgggaa acagttctac aatgggattg agatcaaagt ctgggccatc 9121 gcctgcttcg caccccagaa acagtgtcga gaagaggtgc tcaagaactt cacagaccag 9181 cttcggaaga tttccaagga tgcagggatg cccatccagg gtcagccatg cttctgcaaa 9241 tacgcacagg gggcagacag cgtggagccc atgttccggc atctcaagaa cacctactca 9301 ggactgcagc tcattatcgt catcctgccc gggaagacgc cagtgtatgc tgaagtgaaa 9361 cgtgtcggag atacactctt gggaatggca acacagtgtg tgcaggtgaa gaatgtggtc 9421 aagacgtcac cgcagactct gtccaacctc tgcctcaaga tcaatgtcaa actcggtggc 9481 attaacaaca tcctagtccc acaccagcgc tctgctgttt ttcaacagcc agtgattttc 9541 ctgggagcag atgttacaca tcccccagct ggggatggga agaaaccatc tatcacagca 9601 gtggtaggca gcatggacgc acaccccagc cgatactgtg ccaccgtgcg tgtgcagcgc 9661 ccacggcagg agatcattga agacttgtcc tatatggtgc gcgagctgct tatccagttc 9721 tacaagtcca cccgcttcaa gcccacccgc atcatcttct accgcgacgg cgtccccgag 9781 ggccagctgc cccagattct tcactatgag ctgcttgcca ttcgagatgc atgcatcaaa 9841 ctggaaaaag actaccagcc tgggatcaca tatattgtgg tgcagaagcg ccatcacacc 9901 cgcctgttct gtgctgacaa gaatgagcgg attgggaaga gtggtaacat cccagcaggg 9961 accactgtgg acaccaacat cacccaccca tttgagttcg acttctatct gtgcagccat 10021 gcaggcatcc agggtaccag ccgaccatcc cattattatg tcctttggga tgacaaccgt 10081 ttcacagcag atgaactcca gatcttgaca taccagctgt gccacactta cgtacgatgc 10141 acacgttctg tctctatccc agcacctgcc tactatgctc gcttggtggc tttccgggca 10201 cgataccacc tggtggacaa ggaacatgac agtggagagg ggagccacat atcagggcaa 10261 agcaatgggc gagaccccca ggccctggcc aaagctgtgc aggttcacca ggatactcta 10321 cgcaccatgt acttcgcttg ag //