LOCUS       Exported                3518 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  High Copy number plasmid containing Ple341 (PCP2 MiniPromoter) 
            insert.
ACCESSION   .
VERSION     .
KEYWORDS    pEMS2267
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3518)
  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 3518)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3518
                     /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(1297..1313)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1297..1313)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1310..1332)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1321..1337
                     /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(1345..1375)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1390..1411
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1528..1545)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1699..2287)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1779..1798)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2458..3318)
                     /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     3081..3100
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3319..3423)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3491..3509
                     /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 tccaatggcc ggccagatga aacttccact ggtttccgta aaggcgtctt
      481 ggggagggaa gggaagggga tggggacctc ccagtggtat cccctgcttg ggcactgagg
      541 gaaagccaca gtggctcggg ggaaaaggca gggacgtcct ctccccgcct gcctctgtcc
      601 ccagggagtc tcgcctcctg ttcccacctg gggctagggt gatagaggag aggagatagc
      661 tcaacctggc atttaggtgg tgtgggaaca ggagacccca gagctggagc tagggtggcc
      721 ggtggtgtct gaggcaggtg cagagaggga gggagggaag gggcctttga ctccaacctc
      781 ctttttctgt accgactgca ggtggcagct gccctttcag gagccagtgg gggaacctgg
      841 gtggctgggt ggggacacct gcaagtcctc cctaagccag ctaccaccct acactgttgg
      901 cctcccttct ccaactgtgg ggatgctgct caggcctttt gtgacatcac acctgagagt
      961 ccctggggtc cagtcattgc tgctgggcac agcgaggtcc aagctcaggt cgccctgccc
     1021 cctacccacc atgccagatc cagcatcgtt gtgggcaaac aattatctgg atgatcttta
     1081 tggggcttaa gcttgggtgg gagcagatgg ggcatgagct ggggatttgg ggatgggggg
     1141 aatccacacc cccacgtcct ggacgtttaa aaggccctct ctggcactgg gccggggcag
     1201 aggccagcag aaaagtgact ggagtccagg cgcgccatta tcggatcccg ggcccgtcga
     1261 ctgcagaggc ctgcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     1321 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     1381 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     1441 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     1501 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1561 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     1621 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1681 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1741 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1801 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1861 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1921 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1981 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2041 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2101 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2161 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2221 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2281 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2341 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2401 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2461 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2521 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     2581 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     2641 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     2701 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     2761 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     2821 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     2881 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     2941 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     3001 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     3061 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     3121 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     3181 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     3241 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     3301 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     3361 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     3421 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     3481 aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
