LOCUS       sequence-309560-        4731 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High Copy number plasmid containing Ple349 (PDE6H MiniPromoter) 
            insert.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 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: pEMS2279
FEATURES             Location/Qualifiers
     source          1..4731
                     /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(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(join(2558..2564,2565..2582,2583..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(join(3671..4462,4463..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
//
