LOCUS Exported 7732 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS gfap:GFP-NUC SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7732) AUTHORS Bernardos RL, Barthel LK, Meyers JR, Raymond PA TITLE Late-stage neuronal progenitors in the retina are radial Muller glia that function as retinal stem cells. JOURNAL J Neurosci. 2007 Jun 27;27(26):7028-40. PUBMED 17596452 REFERENCE 2 (bases 1 to 7732) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7732 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..19 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 1..18 /label=SP6 /note="SP6 promoter, forward primer" regulatory 3147..3156 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3153..3869 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 3153..3869 /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 complement(3198..3219) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3459..3478) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3806..3827 /label=EGFP-C /note="EGFP, forward primer" CDS 3894..3914 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 3918..3938 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 3942..3962 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 4105..4226 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4142..4161) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4196..4215 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(4233..4688) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4370..4391) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4582..4601 /label=F1ori-R /note="F1 origin, reverse primer" promoter 4715..4819 /gene="bla" /label=AmpR promoter promoter 4821..5178 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(4844..4864) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 5029..5164 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5091..5110 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5213..6007 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(5267..5286) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5877..5896 /label=Neo-F /note="Neomycin resistance gene, forward primer" regulatory 6207..6216 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 6213..6587 /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" rep_origin 6725..7313 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7214..7233 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7467..7484 /label=L4440 /note="L4440 vector, forward primer" protein_bind 7601..7622 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 7637..7667 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7675..7691 /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 7680..7702 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 7699..7715 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 7699..7715 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" ORIGIN 1 atttaggtga cactatagaa tactcaagct atgcatcaag cttggtaccg agctcggatc 61 cactagtaac ggccgccagt gtgctggaat tccacctttg ggatgtagtt gaacgggaga 121 tcaactgcat gatgctatca agtcaatatg gatcaaaatc tcaggaatat ttccaggacc 181 ttattgaatc tatgccatga aggattaagg cagttctgaa aggggaaatg cggtttaacc 241 cggtaatagc aaggtgtacc taataaagtg gccggtgagt gtatatattg ttattgtttc 301 accttcagaa atgacatatc tagacaatag tttccttttt cttttttaat tattgtactt 361 ctgtaagatt cttcaatgtt caatggtttt cttatgttaa aggcgcagag ctgtagtttt 421 agtgaagtaa cagcattccg ttgaccaaag ctgacagatg aaggagagtc agttctgacc 481 ttgcagttgt cagtcacaat taagcctggt ttggttacaa gttaaacgta gacctgcttg 541 tggtcaggat catcctttaa aagaactaga tttgttttta acaatttgta accggtgtgc 601 tctgaagtgt cttaaataaa gcatatagga agcaaagcag tgaaaaatgt tccttataat 661 ctacgagcat tagcttaaat gtataaacaa aataatatta attgtgtgtt cagatcaaat 721 ataggtattg ttacacctaa aataaaccca ttgctttaaa aaataattaa acacagtgta 781 caatattagc atacttcctc tgacatctct aagtaaaaca agagctttca cgatttactc 841 actgacccat cttacagcac ttctctgtga atttaggcca tcacaaatgc tgtgcccagt 901 tctctgactg ggtttgacag tcgcccatct gtgtccagca gtctgatgag ctcatcctct 961 gtaaatctgg ctggctgccg tgttgttttg tgtctgtccg tgagtctctc tggccttcat 1021 acactcctct gctacttcat agactgttga tgagtgcaga ggccaaacca ttagtgcttt 1081 tctttttttt tagacctcag taatgcacaa caatgcaaaa caagtagatt gagccttttg 1141 ttctttggac tgagcatttg gttagaatag cttgaaggga tagttagaag atattttgaa 1201 gggaaactaa aaacctgtta ccatttgaca tccaaagaag gaaaactaat atgattgagt 1261 ggttacagat ttccaacact tttcaacata tattattttg tgttccacta aagaaacagt 1321 caaatattat actgtcaacc aaagtactgc acttgactca aaaaagctct gcagcaagtt 1381 gcctcttatt ctaggtcgag tcaacctttt tttctctctc tctgtcggtt cattcaaggt 1441 tcagtaaatg atgacagaat tttcagcttt tattttgggt gaactaaccc tttatatatt 1501 tgttgtaagt ttatttttct gtatattata aatatattag caacataatt ctaagctaca 1561 cttgtaccca atcaagggtc acactttatt ttgatggtat gtttgtttga atttaagtta 1621 cattgcatct acatgtcaac taattctcat tagattataa gaagactgtt aggttggggt 1681 tagtgtaagt tgacatgtac ttgcaaagtt tctcagagtc agttaaatgt ctgttttgta 1741 gcagtattaa cagatattaa gcacactgta tattaatact caaatgaacc gtcaaaataa 1801 agtgttaccg taaaaattat ttccacatct caaggtagct ttatgaggaa agttttcttt 1861 aagattagtt tcttcgaaca aacttgcatg ttaaatatgt gctcagatat ggggcacaca 1921 gacaaaatgt ttattggacg aataacccct ttaacagtta accagcattt tttaaaggga 1981 tagtcagaag atattttgaa ggaaactaaa aacctgtaac cattcgacaa tcgaagaagg 2041 taaaagaaat atgatagagt agttacagtt tttcaacact ctttataata tattgtttag 2101 tattcaactg aagaaatagt caaacacact gtaaaaaaag ttggtttaaa ttgactcaac 2161 tttcaactca agtatagtac ttgacttgat ttaagttgag ttaactagac taaactaaaa 2221 gcactaaaag cactgcggca agtcgccttt caattttagg ttgagtaaat tattattatt 2281 tttttttaca gttcaggttc agtaaatgat aacataattt tcagttttgg gtgaactatc 2341 cctttatata tttgttgtaa gttaattttt ctgtaaatta taaattaatt agcaaaaaag 2401 ctgcacttct tcaccattaa ttaattaata catctcaaag tagctttata aggaaagttt 2461 tctttaatta tgtttctttg catgttaaaa ttgtgctcag atatggagca cacaagcaaa 2521 atgtttattg gatgaataac ccctttaaca tttaaccagc atgagttcaa gtaaacaaat 2581 caattcctat gatttttctg ctaaaccgat ataaatcagt gctgctgaag agcatgtgtg 2641 tgatgttctc agatcagcac aaaagaggga gcgccagttt cttttaataa tgaagaatta 2701 cgtaatcacc tcctgtgctc tctgaaacca ttaaactccc agacagaaca aaactgtgac 2761 tgagaaagca gagcaacccc ggagtaactc agaagatctg caaaccaact ctttagcatg 2821 aggtaagaga gaattaaagt ccaatatcag tgctgatgcc atctcagtct ctctatttct 2881 gtgttccagc agggtatgaa taggattttc tctgactcac tgtgaaaaga ggcagtcagg 2941 agactggctg gacaagcaca aacaattgcg ggctcacaaa caccccctcg gcctttgccc 3001 tccccatcag accccccctc tctctctcac cccccacatc aactctataa aaacccagag 3061 ttcacccgca ctcgatctca ttctcctcca ccatggagtc ccagcgttcc tcgtcgacgg 3121 taccgcgggc ccgggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca 3181 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg 3241 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca 3301 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc 3361 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc 3421 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc 3481 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg 3541 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca 3601 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc 3661 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg 3721 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca 3781 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga 3841 tcactctcgg catggacgag ctgtacaagt ccggactcag atcccgagct gatccaaaaa 3901 agaagagaaa ggtagatcca aaaaagaaga gaaaggtaga tccaaaaaag aagagaaagg 3961 taggatccac cggatctaga taagcggccg cgactctaga tcataatcag ccataccaca 4021 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat 4081 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa 4141 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt 4201 ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa 4261 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa 4321 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa 4381 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca 4441 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg 4501 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc 4561 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc 4621 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca 4681 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt 4741 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata 4801 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta 4861 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 4921 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 4981 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta 5041 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca 5101 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga 5161 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga 5221 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga 5281 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg 5341 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga 5401 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt 5461 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct 5521 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct 5581 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg 5641 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca 5701 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga 5761 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt 5821 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt 5881 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct 5941 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt 6001 cttctgaatt attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg 6061 cggtatttca caccgcatca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt 6121 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa 6181 tgcttcaata atagcacgtg aggagggcca ccatggccaa gttgaccagt gccgttccgg 6241 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct 6301 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca 6361 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg 6421 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct 6481 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg 6541 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctaaaac 6601 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa 6661 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat 6721 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc 6781 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg 6841 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc 6901 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg 6961 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg 7021 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa 7081 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg 7141 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga 7201 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct 7261 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca 7321 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc 7381 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg 7441 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc 7501 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca 7561 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc 7621 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga 7681 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag ct //