LOCUS       sequence-157020-        5837 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High copy number plasmid containing Ple304 MiniPromoter insert..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5837)
  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 5837)
  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: pEMS1963
FEATURES             Location/Qualifiers
     source          1..5837
                     /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"
     rep_origin      complement(187..642)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(324..345)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     536..555
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     840..862
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     854..871
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     855..871
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3616..3632)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3616..3632)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3629..3651)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3640..3656
                     /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(3664..3670,3671..3688,3689..3694))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3709..3730
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3847..3864)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4018..4606)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4098..4117)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4777..5568,5569..5637))
                     /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     5400..5419
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5638..5742)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5810..5828
                     /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 accataaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
      241 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaatagc
      301 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      361 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
      421 cacccaaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
      481 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
      541 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
      601 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtactatggt tgctttgacg
      661 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
      721 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
      781 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
      841 ccagtcacga cgttgtaaaa cgacggccag tgaattggag gctacagtca gtggagagga
      901 ctttcactga ctgactgact gcgtctcaac ctggccggcc actgagttaa tatttccagg
      961 tcagagcgcc tccgagagaa aaaggcggtg gccttggagc tctggtctcc ttaactgcac
     1021 cctgggaagt cttcacacgg cctttcttcc cctggtggtc taatctggag tcagatccag
     1081 ccgtcagctc ttacagacgg ggaaggaccg ggagccaacg ggaaaacaac aggtgagctt
     1141 gtgaacaagt ccccttctgt gtctcgggca gggtgtctgc ccgggcaacc gtggggctgc
     1201 actgacttcc agaaccttct gcctctgcag gtgttttagg aacttggcct cagcgcttca
     1261 gtgacccata ttttattacc tttcaggttt ctataattca tcattcactc ctttctgaaa
     1321 acctagaatg aaaaagttta gcagctcagc gccgcggcct ctctccattc tctccccaaa
     1381 cacatcggcc tgtttgttct ctttcatccc cgctcttcct gttgcattct gccggttcaa
     1441 tggggaagga gtgaaaccgg atggctggac ccagtgcggg gaaatgactt cagagccacc
     1501 tttgtttccc tcattccttt gagcgcactt ggccctgcgt ctctatgaaa gggcttgttt
     1561 gaaccattcg gaacatccag caagccagag atgctgcggc cagaaccgct cgcatctttt
     1621 caggctgcct gagactctgg gttaaactgc caaatttttg caacaaaact aaccatgaca
     1681 ttggacattg tgttataaat tagccacagc cttccaagca aattgtctct ttttattgta
     1741 tcagtgtaag cctcggaaac atttggcttc agctactacc ttcagggagg ctcagaggat
     1801 atttttgttc tgccccaaac caactcttca gacaccacct tccttgtcct cacgcttgac
     1861 tagcagacaa tgggttcttc agaagccaca gcatttcagc tgttttggcc ccagaggcca
     1921 caagctgact gcatgtcatt ctccaccagc agagcgtcac ctcggggtag ctccaaacag
     1981 tatcaaccgg tttgtggtga gtggataaac accaggctgg gtgaatgaag tcacaggctg
     2041 agtcatcctc tgcacatggg ggctgaatgg ggctcagtca ggcccagagg agcctccccg
     2101 gcaaggtgct gggggccagg ctctccctgc cagtgaggct ggggtccgtc cccaatagtc
     2161 attcctttgg ccaacagaaa tgataaggca gacaaatgga agatcaatga agcaacatcg
     2221 tggcaataat gcaattggtc ctgccgtagt catgaacatg taaataacaa gacagcaata
     2281 agaagacccc agactctgcc tagaaacaaa atattcagcc tgttgttgtt gtttcttatt
     2341 gcaggatttt gagtggaaaa gtagttccac ttgtcaatca gctacgttgc aggggctaat
     2401 agattctggt gtttagggag gtcaacacgg aatttaattt aacatctcac aatacaccca
     2461 catgcccaga aaatgtggac agcttgatag ggttcactga gccaaagaag caagacccaa
     2521 acagtgcatt aatgtatagg ggaagaatag ttgcaatcag ccccttgtga attggacaga
     2581 gggcttggag atgaagcaat tatcagccat cggggattaa tagtaaagtg tgaataccaa
     2641 gctgtatatg ttaggggcag aatatgttgc ctatagaaaa caacaaagga cgtttcttac
     2701 tattaagtga gattaaacgt aatggcctga tttaatcaca aaggcagaat taaattcaga
     2761 tggcaacttc aaagtggaca caggaacaca ggccactgct tacatttaca tatagctggc
     2821 atagcttggc ctgcaacaca ataggataaa gtttctttgg ccacttaaac aattttctat
     2881 tcttcacctt tttctttttt tctttttttt ttcttttgtg ggtgtgtgtg tgtgtgtgtg
     2941 tgtgtgtgaa cactcactgt atttattccc caattgcatt tagttagtaa gaaggagcta
     3001 agtgttatat ttattcatgg aggctataac tgttaattct tccaaattgt gaaaaaaaag
     3061 cctcataggg tcaataagaa cagagtagtt ataatcagat tgtcaccaaa aaagaaaaag
     3121 aagaaaaagg ttgtttttga gtagacagtt gctggaggaa ggcacgtgag ttgcagcctg
     3181 accagtgcga ccagtcctcc ttcaacagcc ccgcttgcct tgaaaaggcg cccttactcc
     3241 gctcaggggg cctgtcccgc gcggctggca caggcgctca caataacctc ctaaagcggt
     3301 gccagccgca cctccgcgcg gccccggcac aagcagccaa tgaacacgcg gctgcgcccg
     3361 gcctcgcgcc tccattggct gcgccccgcc acccgctgcc ccgcaggttc ccaagccggg
     3421 tttaaagggg tagggcgcgg gccagggccc caccatcgtt tccccgcgcg caggtccgcg
     3481 gggaggggcg gcctgccgac cggcccaccc cagggcgttc ctgaagggcg tcctcggcgc
     3541 gccagcacag tcttcactga ctgactgact ggaaagagga agggctggaa gaggaaggag
     3601 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     3661 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     3721 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3781 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3841 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3901 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3961 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4021 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4081 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4141 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4201 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4261 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4321 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4381 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4441 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4501 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4561 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4621 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4681 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4741 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4801 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4861 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4921 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4981 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5041 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5101 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5161 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5221 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5281 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5341 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5401 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5461 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5521 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5581 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5641 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5701 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     5761 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     5821 cacgaggccc tttcgtc
//
