LOCUS       sequence-309564-        4441 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High Copy number plasmid containing Ple316 (NR2E1 MiniPromoter) 
            insert.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4441)
  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 4441)
  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: pEMS2232
FEATURES             Location/Qualifiers
     source          1..4441
                     /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(2220..2236)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2220..2236)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2233..2255)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2244..2260
                     /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(2268..2274,2275..2292,2293..2298))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2313..2334
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2451..2468)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2622..3210)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2702..2721)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3381..4172,4173..4241))
                     /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     4004..4023
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4242..4346)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4414..4432
                     /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 tgagctcggt acctcgcgaa tgcatctaga tggccggcca atcaaaggaa
      481 aaggagagag aaaccagcga agccctagag agacagggag aaagcctcaa agcaacggag
      541 acagagccgg gccccaccgc acggctggag ggccgccccc acacggagct gccccagccc
      601 agggcctgcg gggccgagtc gcctcccggc agctgagtgg cagcctcggc tgcggaccca
      661 ggcgcagctt cccccagagg ccggcgctat taactcccgg ggcctcctgc ggcggcgagg
      721 cggtggcacg gctcctcccg ctccctcggc ctccaggttc aataaggatt acatcatgga
      781 gcaagtatgc ggggcagcct gagaatcaaa gccgtcagca ccggcggctg gttaactgta
      841 ctgttgctga aacacatcct tcccaccccc tccccgcgcc cctctccctg ccccggccct
      901 ctagaagacc ccatcctcag ccctgacttt ccgtagatat gtgcaaaatg agtaaattac
      961 tcacctcggg ccagatccaa gttttaccca acagaagggg caccggacca agaatgaacc
     1021 aactcacatg gccatgtccg gcgcgcacaa tcacacgcca gcacacagcc acccaatttc
     1081 ttccgcgaat ctatctggca ctctggagag agggggaaaa gcgttttgag aaagccccgt
     1141 cacccctccc cttccttctt gccgtgaaat atacgaattc atttttatta cgagccgcac
     1201 cgtcctcacc atcacgcacg cacagagcca cactcccata ttcacacttt ctaactcgta
     1261 agctccgaca gcgcctgcat tttctttggg agccgcttgg aggttcatta atatcattag
     1321 catttaaccc cctccctctt cccatcccct ccccgcacat ggctgacgtc agaccccgcc
     1381 aggagttggg ggaaaagcta agtgggccag ggacgcccta ttcccctccc cgcggctgcc
     1441 tgtcagagcg cttctggaga tattacaggg gacccagccc gcagcgacag gcacaaagtc
     1501 acggggtaat gaacttcggg gacccttcgc cgctgcgtgc gcggctctcc ccggaaaccc
     1561 ggacctggcc gcctcttccc tcggaagatt tcccagcaat ctagttttcc cactctgcgc
     1621 ttgggttccg gcagcgcgga gcccgtctgc ctctgagact gcggtagtgt tttccttctt
     1681 tccttgggag accagcggtc ggcagagatt gcccacactc tgcatgccta tgtagaggga
     1741 gagatcgaag actgagtgac aggaatgggg aaaaagaggg atttcgctcc gtaggaaggc
     1801 cattttcgtg tctccatctc tgtctttcaa catccctctc ttgctgttct tccttcttcc
     1861 tcagtcttcc tgtccatctc tccatctgtc tgtccatgtg tgtgtccata tcaagcagca
     1921 ttcccagcag ctgcggtttt gcaagagccg ggaagaaact taaggatgct taaatttcca
     1981 ctgttggacg aattctgagc gcccagggag cagcgcagcg cgcgactgac acccacctgt
     2041 cccgcccagg agccttgcag gctggagggc agctggagag cggcggcgcc cggcggcgag
     2101 gcgggcgctg ccggccggga ctcgggcagc gcccaccaac cgctggcgcg ccatcggatc
     2161 ccatcggatc ccgggcccgt cgactgcaga ggcctgcatg caagcttggc gtaatcatgg
     2221 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     2281 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     2341 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     2401 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     2461 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2521 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2581 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2641 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2701 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2761 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2821 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2881 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2941 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3001 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3061 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3121 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3181 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3241 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3301 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3361 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3421 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3481 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3541 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3601 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3661 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3721 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3781 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3841 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3901 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3961 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4021 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4081 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4141 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4201 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     4261 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4321 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     4381 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     4441 c
//
