LOCUS Exported 3527 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION High copy number plasmid containing Ple344 (TUBB3 MiniPromoter) insert.. ACCESSION . VERSION . KEYWORDS pEMS2177 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3527) AUTHORS Simpson E, Korecki AJ, Fornes O, McGill TJ, Cueva-Vargas JL, Agostinone J, Farkas RA, Hickmott JW, Lam SL, Mathelier A, Renner LM, Stoddard J, Zhou M, Di Polo A, Neuringer M, Wasserman WW TITLE New MiniPromoter Ple345 (NEFL) Drives Strong and Specific Expression in Retinal Ganglion Cells of Mouse and Primate Retina. JOURNAL Hum Gene Ther. 2018 Jul 31. doi: 10.1089/hum.2018.118. PUBMED 30062914 REFERENCE 2 (bases 1 to 3527) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3527 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(29..51) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 151..170 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 364..386 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 378..395 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 379..395 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1306..1322) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1306..1322) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1319..1341) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1330..1346 /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(1354..1384) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1399..1420 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1537..1554) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1708..2296) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1788..1807) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2467..3327) /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 3090..3109 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3328..3432) /gene="bla" /label=AmpR promoter primer_bind 3500..3518 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa 421 tgcatctaga tggccggcct cgcagctgct cattttgtga agtgaatgct gatttggggg 481 ccgggtgggg ttcgtctgta catcgtgcac tgtcagaccc ttcctgaagg atttttgtta 541 ctgaagtatc agaaggccct tgttctaagg tggtggcact gtaatatctg gcccaggagg 601 aggtcagtgc cgtgccggcg agtcctttgc tcccttcctt ctctgaccag taaacgcatc 661 tccagtcgct cccccgaggc ccccctgcac gcagtctccg acctcagaga gtgagtcaga 721 tagaagccgc tgcccaccct cccccaccct ggccgggttc ttgcctggct tgtgcgtcct 781 tgtgacaatt cctgacacgg atgcgcaccg ggcagtggca cgtccaggtg ctgtgcgcgc 841 ccgggcccac aaagctcctt tggacgctca tggcagcccc ttggtggagg aggtcgggtg 901 cccaccccct ccccgcccac tgcggaagcc gctcctctgg gagacagccc ctcctttcga 961 atgcgcgggg ccctcagacc gcgcccggcc cagcgctggg ggatccttgg ctgcgggagg 1021 ggcgccgcat tgcgcgcggc gggcggggac gcgcggtgcg gagcctgcgg gccgggcggg 1081 gctctgcggc ggcgcctccc gattggccac ccgcggtgac atcagccgat gcgaagggcg 1141 gggccgcggc tataagagcg cgcggccgcg gtccccgacc ctcagcagcc agcccggccc 1201 gcccgcgccc gtccgcagcc gcccgccaga cgcgcccagt ggcgcgccat cggatcccgg 1261 gcccgtcgac tgcagaggcc tgcatgcaag cttggcgtaa tcatggtcat agctgtttcc 1321 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 1381 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 1441 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 1501 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 1561 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 1621 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 1681 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 1741 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 1801 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 1861 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 1921 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 1981 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 2041 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 2101 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg 2161 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 2221 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 2281 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 2341 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 2401 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 2461 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 2521 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 2581 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 2641 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 2701 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 2761 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 2821 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 2881 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 2941 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 3001 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 3061 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 3121 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 3181 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 3241 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 3301 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 3361 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 3421 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat 3481 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtc //