LOCUS       sequence-215959-        4694 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High copy number plasmid containing Ple320 (POU4F1 MiniPromoter) 
            insert..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4694)
  AUTHORS   Simpson E, Korecki AJ, Fornes O, McGill TJ, Cueva-Vargas JL, 
            Agostinone J, Farkas RA, Hickmott JW, Lam SL, Mathelier A, Renner 
            LM, Stoddard J, Zhou M, Di Polo A, Neuringer M, Wasserman WW
  TITLE     New MiniPromoter Ple345 (NEFL) Drives Strong and Specific Expression
            in Retinal Ganglion Cells of Mouse and Primate Retina.
  JOURNAL   Hum Gene Ther. 2018 Jul 31. doi: 10.1089/hum.2018.118.
  PUBMED    30062914
REFERENCE   2  (bases 1 to 4694)
  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: "Hum Gene 
            Ther. 2018 Jul 31. doi: 10.1089/hum.2018.118."
COMMENT     Sequence Label: pEMS2235
FEATURES             Location/Qualifiers
     source          1..4694
                     /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"
     CDS             1796..1807
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2473..2489)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2473..2489)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2486..2508)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2497..2513
                     /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(2521..2527,2528..2545,2546..2551))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2566..2587
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2704..2721)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2875..3463)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2955..2974)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3634..4425,4426..4494))
                     /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     4257..4276
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4495..4599)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4667..4685
                     /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 tggccggccg aaatacatac actagatttc tccccctttt gtgcctttgg
      481 atggtatttg tagagagaat caaactgaga ttaactgttt atgggatgta ctttattaga
      541 ttagaagggt ttcccagagc ctcataagaa gttctttcat tcccatttgt gggagtccta
      601 caacacagct ttggcctatc acgatgtaac ctgatccttt taataatact tggcagagaa
      661 aaccttttaa gcaatgatct gagtgcacag tgaacattta ttgcctaatt ctcggaaccc
      721 cagtcagcta ggaagcatct gtgaagatgc agatttcttt acagaaccca cctggagtgc
      781 aagttagggc gatcttcagt tttatatgac tgagacagtc tagaatgcct tgagttgcag
      841 ctccacttca gaaacaaggc agattcttag gcctgctatg tctttcctgt acccttcagg
      901 gtccctaaag accagcgctt ccttgagagg ggaggggcag gttccaggcc ctcaccctct
      961 gtgggggcct tttctttccc gtcaagagag agcgcaggga acggccacca ggggtgtcag
     1021 gaaggactag gttaagcccg gactcccggt gtgatgacta ccgtaggact cactctttcc
     1081 ctctagcgac gtcctgctca tcgtccactc cccgttgaga taaagttgca tttagcgctc
     1141 ccaggggact gcgtaatgga tggaaccagc tccttctctc accagcaagt gaacatcctt
     1201 ctcacttccc tctggagcta gaaggaaggg agggggtggg ggggaacatg aaaagaaaga
     1261 aagaaaaagc catggtgacg cctgtgggct cttcttggaa tctgagtagg tgagtgggta
     1321 tgctaaacac acatgcattt gtgcagagga aagataaaat ccacgggaac gcaaatttag
     1381 gtttatagaa atgagatctg ctctttggtg gaaatacaga cttccttggt gtgggagaga
     1441 acagaaagat tatctgattt ctgctttcca taagcagccc cttgctgctc tccagctctc
     1501 ttaccgcaga tgtgataaca tctgggctgt taggttatta tcaccgcgag caagtgctcc
     1561 ttccctgggg actttggcac ctgaactgtg gctcccctgg gaggctcttg atctgctacc
     1621 acataagggc catataacag gttggctaat tgatttattc tttttaggtc ttaatttcac
     1681 aataattttt taaaaattta aaaactggac cagagcttgt ccccgttctt tcaaattggc
     1741 ttgatttaat atttttgaat caagctttcc tgtaagttag tttgatctgt cacacataga
     1801 ggggcgtgcg cgcatctcgt cttttttatt ctaaaaggtg cccaccctta gacaatagct
     1861 ggaccttctg atctttccag accacggtgg gaaaggtgag gcgggcagtg cgcatgcccg
     1921 ggtagctaca ggtgcatcac ctgcgctgtc cttgctggtg gtcactataa gagcggctcc
     1981 cacgcgctgg tcgcggccgg acgagcgcgc gcgcgcgcag acgcacgggg ccgccgagaa
     2041 ggagagcgcg cgtcagacag acggagacag ctgatgcctg tcagcgcggt ggggccgcga
     2101 gctctcgcga gagctccccg ggacggccgc gagaagtggg gctcaggtgt tagaggagtg
     2161 catcccgtcg gcgcgcgggg ctagagctgt cggagaagcg ggaccgcgag gccggcgcgc
     2221 ggcgctctgc gcggtcagag ggagcgcctg gcagcagcag gagcagcagc agcagcccgc
     2281 ggcggggccg ccgccagccg ccgcgaccgc cgcggctgca gcctccgaag ggaggccggg
     2341 tgagccggcg tacgcacttt cccgcggact ttcggagtgt ttgtggatat acatgccaag
     2401 ccgccacggc gcgccatcgg atcccgggcc cgtcgactgc agaggcctgc atgcaagctt
     2461 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     2521 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     2581 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     2641 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     2701 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2761 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2821 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2881 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2941 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3001 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3061 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3121 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3181 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3241 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3301 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3361 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3421 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3481 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3541 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3601 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3661 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3721 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     3781 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3841 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3901 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3961 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4021 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4081 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4141 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4201 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4261 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4321 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     4381 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4441 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     4501 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4561 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     4621 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     4681 gaggcccttt cgtc
//
