LOCUS Exported 7830 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Self-inactivating retrovirus vector containing the DsRed-Express2 gene. ACCESSION . VERSION . KEYWORDS pSIR-DsRed-Express2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7830) AUTHORS Fujita T, Fujii H TITLE Identification of Proteins Associated with an IFNgamma-Responsive Promoter by a Retroviral Expression System for enChIP Using CRISPR. JOURNAL PLoS One. 2014 Jul 22;9(7):e103084. doi: 10.1371/journal.pone.0103084. eCollection 2014. PUBMED 25051498 REFERENCE 2 (bases 1 to 7830) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7830 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 580..937 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 996..1412 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1342..1364 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1380..1396 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" CDS 2815..3492 /codon_start=1 /product="noncytotoxic tetrameric variant of DsRed fluorescent protein (Strack et al., 2008)" /label=DsRed-Express2 /note="mammalian codon-optimized" /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH LFQ" primer_bind complement(2995..3015) /label=DsRed1-N /note="DsRed1, reverse primer" primer_bind 3213..3232 /label=mCherry-F /note="mCherry, forward primer" primer_bind 3407..3430 /label=DsRed1-C /note="DsRed1, forward primer" primer_bind complement(4814..4830) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4814..4830) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4827..4849) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4838..4854 /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(4862..4892) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4907..4928 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5045..5062) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5216..5804) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5296..5315) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5975..6835) /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 6598..6617 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6836..6940) /gene="bla" /label=AmpR promoter primer_bind 7008..7026 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(7064..7086) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 7186..7205 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 7399..7421 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 7413..7430 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 7414..7430 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" ORIGIN 1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat 61 ggaaaaatac ataactgaga atagagaagt tcagatcaag gtcaggaaca gatggaacag 121 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa 181 gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt 241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt 301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac 361 ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa 421 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc 481 tgagtgattg actacccgtc agcgggggtc tttcatttgg gggctcgtcc gggatcggga 541 gacccctgcc cagggaccac cgacccacca ccgggaggta agctggccag caacttatct 601 gtgtctgtcc gattgtctag tgtctatgac tgattttatg cgcctgcgtc ggtactagtt 661 agctaactag ctctgtatct ggcggacccg tggtggaact gacgagttcg gaacacccgg 721 ccgcaaccct gggagacgtc ccagggactt cgggggccgt ttttgtggcc cgacctgagt 781 ccaaaaatcc cgatcgtttt ggactctttg gtgcaccccc cttagaggag ggatatgtgg 841 ttctggtagg agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg 901 tttgggaccg aagccgcgcc gcgcgtcttg tctgctgcag catcgttctg tgttgtctct 961 gtctgactgt gtttctgtat ttgtctgaga atatgggcca gactgttacc actcccttaa 1021 gtttgacctt aggtcactgg aaagatgtcg agcggatcgc tcacaaccag tcggtagatg 1081 tcaagaagag acgttgggtt accttctgct ctgcagaatg gccaaccttt aacgtcggat 1141 ggccgcgaga cggcaccttt aaccgagacc tcatcaccca ggttaagatc aaggtctttt 1201 cacctggccc gcatggacac ccagaccagg tcccctacat cgtgacctgg gaagccttgg 1261 cttttgaccc ccctccctgg gtcaagccct ttgtacaccc taagcctccg cctcctcttc 1321 ctccatccgc cccgtctctc ccccttgaac ctcctcgttc gaccccgcct cgatcctccc 1381 tttatccagc cctcactcct tctctaggcg ccaaacctaa acctcaagtt ctttctgaca 1441 gtggggggcc gctcatcgac ctacttacag aagacccccc gccttatagg gacccaagac 1501 cacccccttc cgacagggac ggaaatggtg gagaagcgac ccctgcggga gaggcaccgg 1561 acccctcccc aatggcatct cgcctacgtg ggagacggga gccccctgtg gccgactcca 1621 ctacctcgca ggcattcccc ctccgcgcag gaggaaacgg acagcttcaa tactggccgt 1681 tctcctcttc tgacctttac aactggaaaa ataataaccc ttctttttct gaagatccag 1741 gtaaactgac agctctgatc gagtctgttc tcatcaccca tcagcccacc tgggacgact 1801 gtcagcagct gttggggact ctgctgaccg gagaagaaaa acaacgggtg ctcttagagg 1861 ctagaaaggc ggtgcggggc gatgatgggc gccccactca actgcccaat gaagtcgatg 1921 ccgcttttcc cctcgagaag cttgtcgacg gatccgaatt cgtcgaggga agaaataata 1981 aagacattaa aatcactttt gtaaactttt taaataaggg aaattgatta aaacaacgcg 2041 aagttgtaaa atgccatata ctgcatacat tttgtgggaa aaaatgtgca tttacctgct 2101 caccaaatta tcttattagc tagcagtaga gctctttgtg tgatgggaag atgggataac 2161 agctgttgtt actgtactgg tttttttttt taatatattt aaaaaaaaga gtgtctccta 2221 aagtgtaagc gctatcatgc tggactaatg ttttaattta atttaattgt tgtcttggta 2281 gagatggggt ctcgctattt tgctcacgca gatctcaaac tcctgacgcc caggtacact 2341 cccatctcgg cctcctaaaa tggtgggatc acagacgtca gccaccgagc ccggtcactt 2401 ttttgtattc cccacagtat tgatgtatat cttctgcgtt caaaagcaat tttttaaagc 2461 ctcataacgt ggtaacagaa tactttgcac attacaaaat tcagaacacg gaaacaagaa 2521 gctcgctttt ttttcccccc tatttcggtt tggcccttta gatttcccct cccccaccgg 2581 ggcgggactt cccgccgact tctttcaggt tctcagttcg gtccgccaac tgtcgtataa 2641 aggcgctgcc tcaggccaga ggcctcacaa agcgcgtcga gacagcaagc gaaccggaat 2701 tgccagctgg ggcgccctct ggtaaggttg ggaagccctg caaagtaaac tggatggctt 2761 tcttgccgcc aaggatctga tggcgcaggg gatcaagatc gatctgatcc aaccatggat 2821 agcactgaga acgtcatcaa gcccttcatg cgcttcaagg tgcacatgga gggctccgtg 2881 aacggccacg agttcgagat cgagggcgag ggcgagggca agccctacga gggcacccag 2941 accgccaagc tgcaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc 3001 ccccagttcc agtacggctc caaggtgtac gtgaagcacc ccgccgacat ccccgactac 3061 aagaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc 3121 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcaccttcat ctaccacgtg 3181 aagttcatcg gcgtgaactt cccctccgac ggccccgtaa tgcagaagaa gactctgggc 3241 tgggagccct ccaccgagcg cctgtacccc cgcgacggcg tgctgaaggg cgagatccac 3301 aaggcgctga agctgaaggg cggcggccac tacctggtgg agttcaagtc aatctacatg 3361 gccaagaagc ccgtgaagct gcccggctac tactacgtgg actccaagct ggacatcacc 3421 tcccacaacg aggactacac cgtggtggag cagtacgagc gcgccgaggc ccgccaccac 3481 ctgttccagt agatggctgg atgatcctcc agcgcgggga tctcatgctg gagatcccgc 3541 gattagtcca atttgttaaa gacaggatat cagtggtcca ggctctagtt ttgactcaac 3601 aatatcacca gctgaagcct atagagtacg agccatagat aaaataaaag attttattta 3661 gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag 3721 taacgccatt ttgcaaggca tggaaaaata cataactgag aatagagaag ttcagatcaa 3781 ggtcaggaac agatggaaca ggtcgaccta gagaaccatc agatgtttcc agggtgcccc 3841 aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc 3901 tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggggc 3961 gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa ccctcttgca 4021 gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta 4081 cccgtcagcg ggggtctctt cacatgcagc atgtatcaaa attaatttgg tttttttttc 4141 ttaagtattt acattaaatg gcccatagta cttaaagtaa catgggcttc cttgaaataa 4201 acatggagta ttcagaatgt gtcataaata tttctaattt taagatagta tctccattgg 4261 ctatctactt tttcttatat ttttttttgt cctctgtctt ccattcgttg ttgttgttgt 4321 ttgtttgttt gtttgttggt tggttggtta attttttttt aaagatccta cactatagtt 4381 caagctagac tattagctac tctgtaaccc agggtgacct ttgaagtcat gggtagcctg 4441 ctgttttagc cttcccacat ctaagattac aggtatgagc tatcattttt ggtatattga 4501 ttgattgatt gattgatgtg tgtgtgtgtg attgtgtttg tgtgtgtgat tgtgtatatg 4561 tgtgtatggt tgtgtgtgat tgtgtgtatg tatgtttgtg tgtgattgtg tgtgtgtgat 4621 tgtgcatgtg tgtgtgtgtg attgtgttta tgtgtatgat tgtgtgtgtg tgtgtgtgtg 4681 tgtgtgtgtg tgtgtgtgtg tgtgtgttgt gtatatatat ttatggtagt gagaggcaac 4741 gctccggctc aggtgtcagg ttggtttttg agacagagtc tttcgcttag cttggaatta 4801 attcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac 4861 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac 4921 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc 4981 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg 5041 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc 5101 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt 5161 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 5221 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa 5281 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc 5341 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg 5401 cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc 5461 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc 5521 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca 5581 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 5641 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 5701 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt 5761 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 5821 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 5881 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa 5941 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac 6001 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 6061 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc 6121 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca 6181 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta 6241 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg 6301 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc 6361 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg 6421 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt 6481 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt 6541 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata 6601 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc 6661 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac 6721 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa 6781 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct 6841 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat 6901 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc 6961 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca 7021 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc 7081 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg 7141 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga 7201 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat 7261 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc 7321 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt 7381 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gccaagctag 7441 ctttgctctt aggagtttcc taatacatcc caaactcaaa tatataaagc atttgacttg 7501 ttctatgccc tagggggcgg ggggaagcta agccagcttt ttttaacatt taaaatgtta 7561 attccatttt aaatgcacag atgtttttat ttcataaggg tttcaatgtg catgaatgct 7621 gcaatattcc tgttaccaaa gctagtataa ataaaaatag ataaacgtgg aaattactta 7681 gagtttctgt cattaacgtt tccttcctca gttgacaaca taaatgcgct gctgagaagc 7741 cagtttgcat ctgtcaggat caatttccca ttatgccagt catattaatt actagtcaat 7801 tagttgattt ttatttttga catatacatg //