LOCUS Exported 5438 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION High copy number plasmid containing Ple343 (TUBB3 MiniPromoter) insert.. ACCESSION . VERSION . KEYWORDS pEMS2176 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5438) 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 5438) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5438 /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(3217..3233) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3217..3233) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3230..3252) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3241..3257 /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(3265..3295) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3310..3331 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3448..3465) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3619..4207) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3699..3718) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4378..5238) /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 5001..5020 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5239..5343) /gene="bla" /label=AmpR promoter primer_bind 5411..5429 /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 ttacctcgcg aatgcatcta gatggccggc ccctgcgctg ggagccgagc 481 ccccggcagc tcgcggggat gcagtaagcc gggcgcagcc cgccgggagg atgagaaaac 541 ccgaagcccg gaaagccgag cggccgcctg cagtcacccc ggggggggca ggggcgggcc 601 gggctatgca gaaacaccgg ggcccgcggg acacaggacg ctgccccact gccgcagagc 661 tgaatctgga cccccgggct ttgctttcct ggggcctgtg ctcctccctc cccggggcgg 721 ggttcgcctg cagcggcgga ggggggagcc tttgtcctcc gcgcgggctg cggcaccgcc 781 tcctcgcggc tgcggggcgg ggctgggggc ggggcctggc cgtctcagcc aatgggaggc 841 ggagtccctg gctcgccccg ccctctcgcc tgaaagacct gcggagccgc gggccgggcg 901 ggaaccgcat tcgggtcctg gaggggcttg gaggccccag tctcgcagcc ccttcatcgg 961 ggacccccgc tcccacctgc cttggccact gccctcaaag cccggccccg ctttccccag 1021 gaccccaagc gctgcgcgag gtcagcacca aggacagcgc ccgggccgag cgcctggcct 1081 cgaacgccgg gttctgcttt gtgtttccag gtttttgttt ttggttatga gaggaggcct 1141 ggctctgtcg cccgggctgg cgtccagagg cgcgatctcg gctcactgca gcctcggcct 1201 cccgagtagc tgggaccgca ggcgctcacc accacgcctg gccacaattt tccttttttc 1261 ttattttctt ttagatggag tttggctctt gttgcccagg ctgggggacg tcgtgtcccc 1321 tggtcctggg aggctgtccc tggcccaagg tcacacagca ggtgcgagag agctgggctg 1381 gcccttggtt ctcatcccct gcaggagcca acctggggct cgggcggggc gggttggggg 1441 cgctgcttgc caaaggatct gccctccgaa cagtgttggg gaacagggga tcgggggggg 1501 aggctggctt tgtcaggacc aaggccaggg acccagctca gggctggtgc agcgctgatt 1561 ctgggaagga gggggtgcag gagacggtcc tggaggggaa ggggccagag aaaggcagcc 1621 tccggaggct gttgccatgg aaaccaggca ggaacaaaaa gctaattaca acagggccag 1681 ccacgtggct gacatgtaaa ttgcaacttt ggctccgggt ggaggcgcct ctgagggtgg 1741 gaggcccagt agtggggaga ctctaatcac ccaaggagac agctgtgcct gccaaggaga 1801 gcctggggga ggggagacct gggggacagt ggcctcaaat caaaccagga ctcccggagc 1861 agcctctgac agctgccttg gcctgatggg aggggctgag ggctcattaa tgtggtcact 1921 ggggccaagc ctcccctgag catctgctcc tgggagggtc aggggtcgct ggggggaggg 1981 tcgcagctgc tcattttgtg aagtgaatgc tgatttgggg gccgggtggg gttcgtctgt 2041 acatcgtgca ctgtcagacc cttcctgaag gatttttgtt actgaagtat cagaaggccc 2101 ttgttctaag gtggtggcac tgtaatatct ggcccaggag gaggtcagtg ccgtgccggc 2161 gagtcctttg ctcccttcct tctctgacca gtaaacgcat ctccagtcgc tcccccgagg 2221 cccccctgca cgcagtctcc gacctcagag agtgagtcag atagaagccg ctgcccaccc 2281 tcccccaccc tggccgggtt cttgcctggc ttgtgcgtcc ttgtgacaat tcctgacacg 2341 gatgcgcacc gggcagtggc acgtccaggt gctgtgcgcg cccgggccca caaagctcct 2401 ttggacgctc atggcagccc cttggtggag gaggtcgggt gcccaccccc tccccgccca 2461 ctgcggaagc cggcgaccca cggagctcgc tctcggccgc cgccacccct ctgtgttcgc 2521 gcccttccga gctctgatcc gacgctttgt ttcttctcag tgggttcagg gcctgggcca 2581 gcctttacct acctccccca cccaaaaccg gcaaaagctc agagcacctt gtctgccaaa 2641 agacagggag ctgggatggt gcgggttggt ctctaaaccg gcgtggggaa aaaagaccct 2701 ccgtacaaag ccgcagggtg gggctgtcgc aagggcggaa ccgagagggt agctgggggc 2761 ggggttccca gggccaagag gggccattgt cctccctgga gcccggcgcc cccacagcca 2821 gctcctctgg gagacagccc ctcctttcga atgcgcgggg ccctcagacc gcgcccggcc 2881 cagcgctggg ggatccttgg ctgcgggagg ggcgccgcat tgcgcgcggc gggcggggac 2941 gcgcggtgcg gagcctgcgg gccgggcggg gctctgcggc ggcgcctccc gattggccac 3001 ccgcggtgac atcagccgat gcgaagggcg gggccgcggc tataagagcg cgcggccgcg 3061 gtccccgacc ctcagcagcc agcccggccc gcccgcgccc gtccgcagcc gcccgccaga 3121 cgcgcccagt ggcgcgccat cggatcccgg gcccgtcgaa tcggatcccg ggcccgtcga 3181 ctgcagaggc ctgcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa 3241 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg 3301 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca 3361 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg 3421 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 3481 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 3541 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 3601 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 3661 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 3721 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 3781 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 3841 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 3901 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 3961 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 4021 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc 4081 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 4141 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 4201 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 4261 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 4321 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta 4381 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 4441 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag 4501 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca 4561 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc 4621 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 4681 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 4741 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 4801 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat 4861 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt 4921 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 4981 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat 5041 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 5101 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 5161 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 5221 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 5281 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 5341 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt 5401 aacctataaa aataggcgta tcacgaggcc ctttcgtc //