LOCUS       sequence-210435-        4761 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High copy number plasmid containing Ple342 (TUBB3 MiniPromoter) 
            insert..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4761)
  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 4761)
  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: pEMS2175
FEATURES             Location/Qualifiers
     source          1..4761
                     /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(2540..2556)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2540..2556)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2553..2575)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2564..2580
                     /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(2588..2594,2595..2612,2613..2618))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2633..2654
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2771..2788)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2942..3530)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3022..3041)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3701..4492,4493..4561))
                     /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     4324..4343
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4562..4666)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4734..4752
                     /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 ttacctcgcg aatgcatcta gatggccggc ccctgcgctg ggagccgagc
      481 ccccggcagc tcgcggggat gcagtaagcc gggcgcagcc cgccgggagg atgagaaaac
      541 ccgaagcccg gaaagccgag cggccgcctg cagtcacccc ggggggggca ggggcgggcc
      601 gggctatgca gaaacaccgg ggcccgcggg acacaggacg ctgccccact gccgcagagc
      661 tgaatctgga cccccgggct ttgctttcct ggggcctgtg ctcctccctc cccggggcgg
      721 ggttcgcctg cagcggcgga ggggggagcc tttgtcctcc gcgcgggctg cggcaccgcc
      781 tcctcgcggc tgcggggcgg ggctgggggc ggggcctggc cgtctcagcc aatgggaggc
      841 ggagtccctg gctcgccccg ccctctcgcc tgaaagacct gcggagccgc gggccgggcg
      901 ggaaccgcat tcgggtcctg gaggggcttg gaggccccag tctcgcagcc ccttcatcgg
      961 ggacccccgc tcccacctgc cttggccact gccctcaaag cccggccccg ctttccccag
     1021 gaccccaagc gctgcgcgag gtcagcacca aggacagcgc ccgggccgag cgcctggcct
     1081 cgaacgccgg gttctgcttt gtgtttccag gtttttgttt ttggttatga gaggaggcct
     1141 ggctctgtcg cccgggctgg cgtccagagg cgcgatctcg gctcactgca gcctcggcct
     1201 cccgagtagc tgggaccgca ggcgctcacc accacgcctg gccacaattt tccttttttc
     1261 ttattttctt ttagatggag tttggctctt gttgcccagg ctgtcgcagc tgctcatttt
     1321 gtgaagtgaa tgctgatttg ggggccgggt ggggttcgtc tgtacatcgt gcactgtcag
     1381 acccttcctg aaggattttt gttactgaag tatcagaagg cccttgttct aaggtggtgg
     1441 cactgtaata tctggcccag gaggaggtca gtgccgtgcc ggcgagtcct ttgctccctt
     1501 ccttctctga ccagtaaacg catctccagt cgctcccccg aggcccccct gcacgcagtc
     1561 tccgacctca gagagtgagt cagatagaag ccgctgccca ccctccccca ccctggccgg
     1621 gttcttgcct ggcttgtgcg tccttgtgac aattcctgac acggatgcgc accgggcagt
     1681 ggcacgtcca ggtgctgtgc gcgcccgggc ccacaaagct cctttggacg ctcatggcag
     1741 ccccttggtg gaggaggtcg ggtgcccacc ccctccccgc ccactgcgga agccggcgac
     1801 ccacggagct cgctctcggc cgccgccacc cctctgtgtt cgcgcccttc cgagctctga
     1861 tccgacgctt tgtttcttct cagtgggttc agggcctggg ccagccttta cctacctccc
     1921 ccacccaaaa ccggcaaaag ctcagagcac cttgtctgcc aaaagacagg gagctgggat
     1981 ggtgcgggtt ggtctctaaa ccggcgtggg gaaaaaagac cctccgtaca aagccgcagg
     2041 gtggggctgt cgcaagggcg gaaccgagag ggtagctggg ggcggggttc ccagggccaa
     2101 gaggggccat tgtcctccct ggagcccggc gcccccacag ccagctcctc tgggagacag
     2161 cccctccttt cgaatgcgcg gggccctcag accgcgcccg gcccagcgct gggggatcct
     2221 tggctgcggg aggggcgccg cattgcgcgc ggcgggcggg gacgcgcggt gcggagcctg
     2281 cgggccgggc ggggctctgc ggcggcgcct cccgattggc cacccgcggt gacatcagcc
     2341 gatgcgaagg gcggggccgc ggctataaga gcgcgcggcc gcggtccccg accctcagca
     2401 gccagcccgg cccgcccgcg cccgtccgca gccgcccgcc agacgcgccc agtggcgcgc
     2461 catcggatcc cgggcccgtc gaatcggatc ccgggcccgt cgactgcaga ggcctgcatg
     2521 caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     2581 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     2641 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     2701 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2761 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2821 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2881 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2941 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3001 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3061 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3121 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3181 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3241 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3301 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3361 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3421 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3481 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3541 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3601 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3661 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3721 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3781 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3841 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3901 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     3961 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4021 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4081 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4141 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4201 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4261 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4321 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4381 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4441 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4501 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4561 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4621 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     4681 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     4741 gtatcacgag gccctttcgt c
//
