LOCUS Exported 6283 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses GFP from the Gfa2 promoter. This plasmid is a new version of Plasmid 22672 and includes a stop codon in the myrEGFP insert.. ACCESSION . VERSION . KEYWORDS pGFAP::MyrGFP_v2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6283) AUTHORS Nam HS, Benezra R TITLE High levels of Id1 expression define B1 type adult neural stem cells. JOURNAL Cell Stem Cell. 2009 Nov 6. 5(5):515-26. PUBMED 19896442 REFERENCE 2 (bases 1 to 6283) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6283 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 1..589 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 490..509 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 743..760 /label=L4440 /note="L4440 vector, forward primer" protein_bind 877..898 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 913..943 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 951..967 /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 956..978 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 975..991 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 975..991 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 3373..4089 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" CDS 3373..4086 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(3415..3436) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3676..3695) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4023..4044 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(4658..4675) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4658..4674) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4667..4689) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(4883..4902) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5002..5024 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5062..5080) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5148..5252 /gene="bla" /label=AmpR promoter CDS 5253..6113 /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(5471..5490) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" ORIGIN 1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt 181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc 661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg 721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat 781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt 841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta 901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg 961 ataacaattt cacacaggaa acagctatga catgattacg aattcgagct cggtaccaga 1021 tctgagctcc cacctccctc tctgtgctgg gactcacaga gggagacctc aggaggcagt 1081 ctgtccatca catgtccaaa tgcagagcat accctgggct gggcgcagtg gcgcacaact 1141 gtaattccag cactttggga ggctgatgtg gaaggatcac ttgagcccag aagttctaga 1201 ccagcctggg caacatggca agaccctatc tctacaaaaa aagttaaaaa atcagccacg 1261 tgtggtgaca cacacctgta gtcccagcta ttcaggaggc tgaggtgagg ggatcactta 1321 aggctgggag gttgaggctg cagtgagtcg tggttgcgcc actgcactcc agcctgggca 1381 acagtgagac cctgtctcaa aagacaaaaa aaaaaaaaaa aaaaaaaaga acatatcctg 1441 gtgtggagta ggggacgctg ctctgacaga ggctcggggg cctgagctgg ctctgtgagc 1501 tggggaggag gcagacagcc aggccttgtc tgcaagcaga cctggcagca ttgggctggc 1561 cgccccccag ggcctcctct tcatgcccag tgaatgactc accttggcac agacacaatg 1621 ttcggggtgg gcacagtgcc tgcttcccgc cgcaccccag cccccctcaa atgccttccg 1681 agaagcccat tgagcagggg gcttgcattg caccccagcc tgacagcctg gcatcttggg 1741 ataaaagcag cacagccccc taggggctgc ccttgctgtg tggcgccacc ggcggtggag 1801 aacaaggctc tattcagcct gtgcccagga aaggggatca ggggatgccc aggcatggac 1861 agtgggtggc agggggggag aggagggctg tctgcttccc agaagtccaa ggacacaaat 1921 gggtgagggg actgggcagg gttctgaccc tgtgggacca gagtggaggg cgtagatgga 1981 cctgaagtct ccagggacaa cagggcccag gtctcaggct cctagttggg cccagtggct 2041 ccagcgtttc caaacccatc catccccaga ggttcttccc atctctccag gctgatgtgt 2101 gggaactcga ggaaataaat ctccagtggg agacggaggg gtggccaggg aaacggggcg 2161 ctgcaggaat aaagacgagc cagcacagcc agctcatgtg taacggcttt gtggagctgt 2221 caaggcctgg tctctgggag agaggcacag ggaggccaga caaggaaggg gtgacctgga 2281 gggacagatc caggggctaa agtcctgata aggcaagaga gtgccggccc cctcttgccc 2341 tatcaggacc tccactgcca catagaggcc atgattgacc cttagacaaa gggctggtgt 2401 ccaatcccag cccccagccc cagaactcca gggaatgaat gggcagagag caggaatgtg 2461 ggacatctgt gttcaaggga aggactccag gagtctgctg ggaatgaggc ctagtaggaa 2521 atgaggtggc ccttgagggt acagaacagg ttcattcttc gccaaattcc cagcaccttg 2581 caggcactta cagctgagtg agataatgcc tgggttatga aatcaaaaag ttggaaagca 2641 ggtcagaggt catctggtac agcccttcct tccctttttt tttttttttt ttgtgagaca 2701 aggtctctct ctgttgccca ggctggagtg gcgcaaacac agctcactgc agcctcaacc 2761 tactgggctc aagcaatcct ccagcctcag cctcccaaag tgctgggatt acaagcatga 2821 gccaccccac tcagcccttt ccttcctttt taattgatgc ataataattg taagtattca 2881 tcatggtcca accaaccctt tcttgaccca ccttcctaga gagagggtcc tcttgcttca 2941 gcggtcaggg ccccagaccc atggtctggc tccaggtacc acctgcctca tgcaggagtt 3001 ggcgtgccca ggaagctctg cctctgggca cagtgacctc agtggggtga ggggagctct 3061 ccccatagct gggctgcggc ccaaccccac cccctcaggc tatgccaggg ggtgttgcca 3121 ggggcacccg ggcatcgcca gtctagccca ctccttcata aagccctcgc atcccaggag 3181 cgagcagagc cagagcaggt tggagaggag acgcatcacc tccgctgctc gcggggatcc 3241 accatgggtt gctgtttctc caagaccgca gcgaagggag aagccgccgc ggagaggcct 3301 ggggaggcgg ctgtggcctc gtcgccttcc aaagcgaacg gacaggagaa tggccacgtg 3361 aaggtaggat ccgtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc 3421 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat 3481 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc 3541 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac 3601 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc 3661 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc 3721 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc 3781 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag 3841 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg 3901 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc 3961 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat 4021 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg 4081 tacaagtaag gatccgcaga tcccggccag ataccgatgc tgccgcagca aaagcaggag 4141 cagatgccgc cgtcgcaggc gaagatgtcg cagacggagg aggcgatgct gccggcggag 4201 gaggcgaagt aagtagaggg ctgggctggg ctgtgggggg tgtggggtgc gggactgggc 4261 agtctgggag tccctctcac cacttttctt acctttctag gatgctgccg tcgccgccgc 4321 tcatacacca taaggtgtaa aaaatactag atgcacagaa tagcaagtcc atcaaaactc 4381 ctgcgtgaga attttaccag acttcaagag catctcgcca catcttgaaa aatgccaccg 4441 tccgatgaaa aacaggagcc tgctaaggaa caatgccacc tgtcaataaa tgttgaaaac 4501 tcatcccatt cctgcctctt ggtccttggg cttggggagg ggtgcgcgga tgtggttagg 4561 gaacatgact ggtcaaatgg gaagggcttc aaaagaattc ccaatattga ctaccaagcc 4621 acctgtacag atctgcctgc aggcatgcaa gcttggcact ggccgtcgtt ttacaacgtc 4681 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg 4741 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc 4801 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac 4861 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc 4921 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt 4981 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac 5041 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga 5101 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta 5161 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat 5221 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc 5281 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga 5341 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca 5401 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt 5461 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg 5521 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc 5581 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata 5641 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt 5701 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag 5761 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca 5821 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg 5881 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg 5941 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag 6001 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg 6061 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag 6121 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga 6181 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt 6241 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttc //