LOCUS Exported 7882 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Self-inactivating retrovirus vector containing the GFP gene. ACCESSION . VERSION . KEYWORDS pSIR-GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7882) 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 7882) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7882 /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" regulatory 2814..2823 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2820..3539 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 2820..3536 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2865..2886) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3126..3145) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3473..3494 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(4866..4882) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4866..4882) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4879..4901) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4890..4906 /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(4914..4944) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4959..4980 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5097..5114) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5268..5856) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5348..5367) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6027..6887) /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 6650..6669 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6888..6992) /gene="bla" /label=AmpR promoter primer_bind 7060..7078 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(7116..7138) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 7238..7257 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 7451..7473 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 7465..7482 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 7466..7482 /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 atcgccacca 2821 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg 2881 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg 2941 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc 3001 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc 3061 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct 3121 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg 3181 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca 3241 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg 3301 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg 3361 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact 3421 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc 3481 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaaa 3541 gcggccggct ggatgatcct ccagcgcggg gatctcatgc tggagatccc gcgattagtc 3601 caatttgtta aagacaggat atcagtggtc caggctctag ttttgactca acaatatcac 3661 cagctgaagc ctatagagta cgagccatag ataaaataaa agattttatt tagtctccag 3721 aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca 3781 ttttgcaagg catggaaaaa tacataactg agaatagaga agttcagatc aaggtcagga 3841 acagatggaa caggtcgacc tagagaacca tcagatgttt ccagggtgcc ccaaggacct 3901 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg 3961 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggg gcgccagtcc 4021 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc 4081 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag 4141 cgggggtctc ttcacatgca gcatgtatca aaattaattt ggtttttttt tcttaagtat 4201 ttacattaaa tggcccatag tacttaaagt aacatgggct tccttgaaat aaacatggag 4261 tattcagaat gtgtcataaa tatttctaat tttaagatag tatctccatt ggctatctac 4321 tttttcttat attttttttt gtcctctgtc ttccattcgt tgttgttgtt gtttgtttgt 4381 ttgtttgttg gttggttggt taattttttt ttaaagatcc tacactatag ttcaagctag 4441 actattagct actctgtaac ccagggtgac ctttgaagtc atgggtagcc tgctgtttta 4501 gccttcccac atctaagatt acaggtatga gctatcattt ttggtatatt gattgattga 4561 ttgattgatg tgtgtgtgtg tgattgtgtt tgtgtgtgtg attgtgtata tgtgtgtatg 4621 gttgtgtgtg attgtgtgta tgtatgtttg tgtgtgattg tgtgtgtgtg attgtgcatg 4681 tgtgtgtgtg tgattgtgtt tatgtgtatg attgtgtgtg tgtgtgtgtg tgtgtgtgtg 4741 tgtgtgtgtg tgtgtgtgtt gtgtatatat atttatggta gtgagaggca acgctccggc 4801 tcaggtgtca ggttggtttt tgagacagag tctttcgctt agcttggaat taattcgtaa 4861 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 4921 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 4981 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa 5041 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg 5101 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag 5161 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 5221 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 5281 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 5341 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 5401 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 5461 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 5521 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 5581 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 5641 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 5701 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 5761 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 5821 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg 5881 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca 5941 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt 6001 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca 6061 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg 6121 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca 6181 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt 6241 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt 6301 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca 6361 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 6421 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga 6481 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact 6541 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga 6601 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg 6661 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc 6721 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga 6781 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat 6841 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt 6901 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 6961 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac 7021 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc 7081 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag 7141 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca 7201 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg 7261 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc 7321 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct 7381 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg 7441 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct agctttgctc 7501 ttaggagttt cctaatacat cccaaactca aatatataaa gcatttgact tgttctatgc 7561 cctagggggc ggggggaagc taagccagct ttttttaaca tttaaaatgt taattccatt 7621 ttaaatgcac agatgttttt atttcataag ggtttcaatg tgcatgaatg ctgcaatatt 7681 cctgttacca aagctagtat aaataaaaat agataaacgt ggaaattact tagagtttct 7741 gtcattaacg tttccttcct cagttgacaa cataaatgcg ctgctgagaa gccagtttgc 7801 atctgtcagg atcaatttcc cattatgcca gtcatattaa ttactagtca attagttgat 7861 ttttattttt gacatataca tg //