LOCUS       sequence-157019-        5667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High copy number plasmid containing Ple303 MiniPromoter insert..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5667)
  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 5667)
  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: pEMS1962
FEATURES             Location/Qualifiers
     source          1..5667
                     /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"
     RBS             2717..2728
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(3446..3462)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3446..3462)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3459..3481)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3470..3486
                     /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(3494..3500,3501..3518,3519..3524))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3539..3560
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3677..3694)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3848..4436)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3928..3947)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4607..5398,5399..5467))
                     /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     5230..5249
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5468..5572)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5640..5658
                     /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 gggtgaactc tgtggtgact
      961 gctttaatta atttatgtaa ttggaatttc accaagacat aaatttactc atatctattt
     1021 ttaaagacag gaatattgat cttggaccag caataaattc ctgtaatatt tgtcctcagt
     1081 aaaaagccaa gacaattcta ggccatttct ttttgtcttg aaaacgttat gccaaaatca
     1141 tcattttgtt ggctgcatac atcctgattt atattcgatg agagttataa aaggctttta
     1201 tagtggagat ggccgagtga cattttccta taaatcacga tcatttttac tgctacaaag
     1261 ttctaggact gaacttcaaa tacaaccgtg ctgtaaactc tgtggatgga accacatact
     1321 tgctgaatga gaaacatgtt tccctttcac catgagtgac tgcatattcc agtcagtctc
     1381 tggcagatga gagcatcact aaaacccagg gaaccatgct cagctttgat gaagcagctt
     1441 gtgaaacgca ggtcagaaga acttaccatc ttgggaaagt gataaagctg gatggtctct
     1501 aacatttacc taaacttgaa ttggctattt tttgtacata cattttccat ttcaactatt
     1561 agcttagaag tttacaaaag aacagtccag tttttttttt tctattataa cagatacagc
     1621 attaccattt gtaatcgttg atcaaggcat aagcaatttc ttgcttgtca atttgaacat
     1681 ctagccagga ataaatagca cacagtatgt gtaatgcaat cttcaaagcg ttttattgga
     1741 acagttctta gacctcatac aaggaaataa aaaaataaga aatgtaaatg aatgtgatgt
     1801 cagactgagg aagcaagtct ctagagatcc ctggaagtta gaatggattt taaaattcat
     1861 tatgattgaa ttataaaata aatgaaaacc agcagggtta aatatgaggt tgatgggagc
     1921 aagaaaagaa agttatttgg atacccatgg cagggagcat gcaagttcag gttcacctct
     1981 agggcaagga agaattttct tcaaggttct tctctttcag tgtttactct gtagcaggca
     2041 ctgtgactgt gtaagtggca cacccttaag atgcccccaa gttactttgc ccgccttggt
     2101 ggcccccatt tggtcaccgg gctcactgcg tcttctgtcc cagctgagtg gtttctcctt
     2161 gtctcgcctg ccttcagtgt taatttccag aaaaattgaa agaggagggg ttaagttaaa
     2221 catacactgt gagttgttct tacgctacag ttgaagagag atcttgaaat atatttttca
     2281 ttcattttct ctctttgtgt ctcccctatg tgagaaatat ggcatgcaca attggcgcct
     2341 atattatcct ctagccaata ctttatccca gttgccttag caactgtcaa tatttgtctg
     2401 gtatgtcaca gccccagttg gctgatggct ggttgaacat ctacttgtaa agtttaggaa
     2461 agagagacac ccagtggtct aaattaactg agttgctgaa agcctgcttc attttatttt
     2521 cacttcagaa atgtttatta atttttaaat ctgatcttct gttaatcatc aacaatcccc
     2581 acaattaaac ttatgaaggg agaaaagaac atgtgttggc acacttactt tttctgtaaa
     2641 gtcccagaca gtaaatattt taggctttgc ggtccccctg ccactcccct ccccccaaca
     2701 cacacacact tttctctttc tcctctttct ctcctttcct cccttgcttc tctcccctcc
     2761 ctctcaacac attcaatgag tgccctaaac ggtgacaaac ttgcatgtgc ttccctcatg
     2821 actaaacccc tgggccttct gccaatcccc tgcagatggc cccaaactgc aggcatcccg
     2881 taaggggacc ccacgcttgc agccctggtt ggaacggtca gggtggagga ggatggtggg
     2941 gagtggtggt gtcttcgtcc tgggagaagg cgaagcaact tccaggagga aacgggcgtt
     3001 tccttcccac gcgctcgagc gagccctggg tcctggcctc ggaactccac ccagcccctc
     3061 cccaccctct gggaaaagcc agtcgccaca cacaggcaca cgcaggcccc ggcgccgcgc
     3121 cctaaggaga gcagcaccca cagccaattg ccatggcaac cccggggttc gttccacttc
     3181 cccacccagc cgatctcccc cctcctccct gcactgcagc caaccggctt gtgcgcgtcc
     3241 caggagcgcg ctataaaacc tgtgctgggc gtgatcggca agcaccggac cagggggaag
     3301 gcgagcagtg ccaatctaca gcgaagaaag tctcgtttgg taaaagcgag aggggaaagc
     3361 ctgagcggcg cgccacgaag agacgactga ctgactgact ggaaagagga agggctggaa
     3421 gaggaaggag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     3481 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     3541 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     3601 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3661 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3721 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3781 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3841 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3901 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3961 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4021 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4081 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4141 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4201 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4261 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     4321 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4381 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4441 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4501 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4561 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4621 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4681 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4741 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4801 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4861 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4921 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4981 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5041 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5101 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5161 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5221 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5281 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5341 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5401 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5461 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5521 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     5581 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     5641 ataggcgtat cacgaggccc tttcgtc
//
