LOCUS       sequence-309570-        4689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High Copy number plasmid containing Ple331 (PAX6 MiniPromoter) 
            insert.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4689)
  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 4689)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene Ther. 
            2021 Feb 2. pii: 10.1038/s41434-021-00227-z. doi: 
            10.1038/s41434-021-00227-z."
COMMENT     Sequence Label: pEMS2166
FEATURES             Location/Qualifiers
     source          1..4689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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(2468..2484)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2468..2484)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2481..2503)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2492..2508
                     /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(join(2516..2522,2523..2540,2541..2546))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2561..2582
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2699..2716)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2870..3458)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2950..2969)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3629..4420,4421..4489))
                     /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     4252..4271
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4490..4594)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4662..4680
                     /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 accggccggc
      421 ctcccttttg ctacaggcat agtttaagtc ttgtgtcaca aataaaagga tcctccctag
      481 catatggagt agttgccatg tcctcccagg gatggtctgt agctatgttc acctcaggct
      541 catcaagaag aggtaaactg tcaagtataa tttcagtcat agggtggttt ggttgagaca
      601 gattagtggt atgcaatcat gatagatagt ttatcccagc tccaaactgc cgcttgggaa
      661 atcttgaact caaatgaaca gagactcagt cattattagt gtcaaaaagg tatacagagg
      721 caaaatttgg ttggcagcat aaaagatgaa tcaaagcaaa aaagtgacca attgcatgtt
      781 caaaatcgaa attgagatgc atactacagc ttggaaatac tttaaatacc agaaaagtca
      841 cacattatta attgcgcatt gattaatatt taaaacatgt ttctgtaatg tattttcaaa
      901 gagacaataa aattacatgt attttcttat ttcaaaacct ttattctcaa ggtgatgcat
      961 agtaaggagt acccttttag atacaggagt aagatagcaa aatgtttttt aaaaatttgt
     1021 ttaattttta gtatctttcc ttcagctgca tatttccaga aataagactt tttagaagaa
     1081 gaatttagtt ttatgggact attttaatat tcaaatgtct tacagtttga tgactttctc
     1141 aggaagaaat caggttataa taaaagcatg aatttaaaat gtggaagaac ctaatctaaa
     1201 taactttcca aatattttct aaacactgaa aaatatatat ttataaattg taaaatacat
     1261 tattctatta gcaaacacaa actttggtat ctttaaatca gactcccccc ccaaccaaat
     1321 atccacaaaa agaatgtcct caagtgtcaa tgcatgaagt catgcgtgaa gataaatcta
     1381 acagctcttt ctgtgtcatt cctaatgcac ttgtcactca gtattatcca tcctcattaa
     1441 tgacaatggg aaagtttatc ttgggaacac gttttaaatc atctactctt tacaatggga
     1501 ccatgaaatc tcttacaaaa catgaggcgg gaactactct gacaacaaaa ccccttcctg
     1561 gcagctttat tcttaattcc tgttcaagga gtaatctgtt tattataaaa tcatacagaa
     1621 caatatatta aacagaattt gaaacaagaa tagctagtgt gatttccaag cttacaagta
     1681 tttttacaac atttttatta ggctgaaaac actacattga gttctttttg tgtgatttca
     1741 tgcaatcttc tattctacat cgtaaacacc tttgaaaaga agttttgttt tacataattt
     1801 aaaaataatt ttcaaatggt aaggacactg gtgtttatat aattatttta aaatacaata
     1861 acttacatta gcttaaataa aagcagttat taagtaaaaa tgtgctcatg tttaaatttc
     1921 ctaaccaaaa tgccaaaata ttccaactct tggggatcgg aggaggggac agggtgatta
     1981 cccagagagg tagctggcca gcctaagggc agagatcttg gggccctagt gcccgaaggt
     2041 gcggaggagc gcactcggca agactagttt cctggggatc gactctacgc catacaggac
     2101 ggcggcccag gctggaccgg gccgggctag agcagtcaca ggccgggcca aggaaggcca
     2161 aagcaggggt tggagccggc cggaccctgg gtggggagaa gcaggctccc gcccggccgg
     2221 aaaactagtc ggcgcagagc tgtgcccaac tctagccgcc atgacgtcac gcgggccggg
     2281 cagccaatga ggacggcgct ggcgtggata ttaaggaaag ttagcgcctg cctgagcacc
     2341 ctcttttctt atcattgaca tttaaactct ggggcaggtc ctcgcgtaga acgcggctgt
     2401 caggcgcgcc atcggatccc gggcccgtcg actgcagagg cctgcatgca agcttggcgt
     2461 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     2521 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     2581 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     2641 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     2701 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     2761 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     2821 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2881 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2941 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3001 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3061 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3121 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3181 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3241 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3301 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3361 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3421 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3481 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     3541 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3601 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3661 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     3721 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     3781 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3841 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3901 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     3961 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     4021 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     4081 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     4141 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     4201 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     4261 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     4321 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4381 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     4441 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     4501 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     4561 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     4621 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     4681 cctttcgtc
//
