LOCUS Exported 4731 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION High Copy number plasmid containing Ple349 (PDE6H MiniPromoter) insert. ACCESSION . VERSION . KEYWORDS pEMS2279 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4731) AUTHORS Korecki AJ, Cueva-Vargas JL, Fornes O, Agostinone J, Farkas RA, Hickmott JW, Lam SL, Mathelier A, Zhou M, Wasserman WW, Di Polo A, Simpson EM TITLE Human MiniPromoters for ocular-rAAV expression in ON bipolar, cone, corneal, endothelial, Muller glial, and PAX6 cells. JOURNAL Gene Ther. 2021 Feb 2. pii: 10.1038/s41434-021-00227-z. doi: 10.1038/s41434-021-00227-z. PUBMED 33531684 REFERENCE 2 (bases 1 to 4731) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4731 /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(2510..2526) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2510..2526) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2523..2545) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2534..2550 /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(2558..2588) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2603..2624 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2741..2758) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2912..3500) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2992..3011) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3671..4531) /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 4294..4313 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4532..4636) /gene="bla" /label=AmpR promoter primer_bind 4704..4722 /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 taattcataa gtttttttcc aagattaatg ttgttctgga 481 gaatcaaacg tagggtattt agggaaagag gggccaggct tgtacattcc tttttcacgt 541 ttagtgtgtt cccctctgga gtccactatg cattagtcag agtggaaagt tcagtctagg 601 agattccaaa tgtggaggaa atgtgacttt aatgtgtctt cccactcact ggtattttaa 661 gaagcactaa agaaagatta gagattagcg cttgcataag cagctaaagg acagctaaat 721 ttttaattac tttctaatta cttagtacct gcactctact ttccaatata taattaataa 781 cacttttcca tctgtcagaa gggatcaaga accagaagtt atagagctat ggaatacaga 841 tttaagaaaa taattttatt atatttagtc ataaaatttg catacagtag agcttactcc 901 ttgtgtggta tgtaatagta aaggaagatg tgaatggtaa tggagataca gaggagggag 961 cagagggact catccagaga ccaaacactt tcctggcttt gcaaggtaat cattgctttc 1021 aaaactcaga ggaattttgg taaagaccag tggagagcat atgaatgact ctcaagctgt 1081 ctagttgctc atgtaatctg cttagggctg caggttaaga acctctccct gtctagaaag 1141 gataaattga ccttctttct ctgtaatgtt atggagtgta attttctaat aggtaacaac 1201 tggggaaaga acattgaaat tcactggcaa ggcatggaag tgaccagcaa acagacatct 1261 tggtctctgt ctgctttgat gtagtgcagc acaaagacaa ttgaatatgc aaggtacttc 1321 cagatttggg ccctgttcct ggctccacta ttaattttat ttgcagtctt gaatagctga 1381 acttctggtg aatttgagca ggataattga agggctttgg ggggttgatc aggttccaaa 1441 agaaaaattg cagaagaaaa catgttggac ctgtgggcct gggctgtggc tggggatgaa 1501 aatgggaaag aacatgaagg agatacatgc agaccttctt gttatgccat gggacaagca 1561 aataagtgaa acttgattat tacaaggtca ctgcaaaact tgattactga attcatttag 1621 aaggagccca atagtgacct ccctgtttgc agcttcatag taaacagttt tctgaagaag 1681 gaggaatctg tggcactaaa taaccacaga gaaaacagtt ctttactccc aggatgtgcc 1741 tgcaaacttg gcccaggatg gtgtgagtgt gaggaaaagg agaaggggag ctgagagcat 1801 gagaatgacc acagggcagg gcaggtggct gagccttctc cctttgaggt caagtgtttg 1861 ttccattctg aagcagggtg ctggctggaa ttttggtaat ttttttttta tggctttggt 1921 ttgggtcttg ctttgctttt caaattattg attttctctg cctcagcagg accctgggtg 1981 atatttatgg gctcaacccc agcagtactt cccttcccag ctcctttcac tgtgtatatg 2041 tgtgtgtgtg attgcgcaca taggtgtgtg tttgtcagag aaagggaaga aatgggggta 2101 ttgaaatcta aaggtaagaa taaaaaaaag acagaagata ttgaatcaca aactgaaatt 2161 tacccttgga tatcaacact ttttgtgttt ttgtgaatac aatacctatt tattgactgg 2221 aatgaaccca agtcagtttt gaaagggcct ctaattattg gaaaactcct aagcctgtcc 2281 cagcatcaac ccatgttctc ctaacttctt tgcacctggg ttgctgttct gagcttatga 2341 cggtgagatt aggcagcatg agggtattac agagagataa gtcaggatgc aaagatcctt 2401 ttgtcttttg caaagaaggc agctgggcag ctgataggaa caacggcgcg ccatcggatc 2461 ccgggcccgt cgactgcaga ggcctgcatg caagcttggc gtaatcatgg tcatagctgt 2521 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 2581 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 2641 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 2701 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 2761 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 2821 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 2881 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 2941 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 3001 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 3061 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 3121 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 3181 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 3241 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 3301 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 3361 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 3421 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 3481 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 3541 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 3601 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 3661 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 3721 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 3781 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 3841 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 3901 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 3961 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 4021 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 4081 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 4141 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 4201 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 4261 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 4321 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 4381 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 4441 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 4501 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 4561 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 4621 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta 4681 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt c //