LOCUS       sequence-215960-        4538 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High copy number plasmid containing Ple321 (TUBB3 MiniPromoter) 
            insert..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4538)
  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 4538)
  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: pEMS2236
FEATURES             Location/Qualifiers
     source          1..4538
                     /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(2317..2333)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2317..2333)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2330..2352)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2341..2357
                     /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(2365..2371,2372..2389,2390..2395))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2410..2431
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2548..2565)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2719..3307)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2799..2818)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3478..4269,4270..4338))
                     /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     4101..4120
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4339..4443)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4511..4529
                     /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 ggccggccag gggacgtcgt gtcccctggt cctgggaggc tgtccctggc
      481 ccaaggtcac acagcaggtg cgagagagct gggctggccc ttggttctca tcccctgcag
      541 gagccaacct ggggctcggg cggggcgggt tgggggcgct gcttgccaaa ggatctgccc
      601 tccgaacagt gttggggaac aggggatcgg ggggggaggc tggctttgtc aggaccaagg
      661 ccagggaccc agctcagggc tggtgcagcg ctgattctgg gaaggagggg gtgcaggaga
      721 cggtcctgga ggggaagggg ccagagaaag gcagcctccg gaggctgttg ccatggaaac
      781 caggcaggaa caaaaagcta attacaacag ggccagccac gtggctgaca tgtaaattgc
      841 aactttggct ccgggtggag gcgcctctga gggtgggagg cccagtagtg gggagactct
      901 aatcacccaa ggagacagct gtgcctgcca aggagagcct gggggagggg agacctgggg
      961 gacagtggcc tcaaatcaaa ccaggactcc cggagcagcc tctgacagct gccttggcct
     1021 gatgggaggg gctgagggct cattaatgtg gtcactgggg ccaagcctcc cctgagcatc
     1081 tgctcctggg agggtcaggg gtcgctgggg ggagggttcg cagctgctca ttttgtgaag
     1141 tgaatgctga tttgggggcc gggtggggtt cgtctgtaca tcgtgcactg tcagaccctt
     1201 cctgaaggat ttttgttact gaagtatcag aaggcccttg ttctaaggtg gtggcactgt
     1261 aatatctggc ccaggaggag gtcagtgccg tgccggcgag tcctttgctc ccttccttct
     1321 ctgaccagta aacgcatctc cagtcgctcc cccgaggccc ccctgcacgc agtctccgac
     1381 ctcagagagt gagtcagata gaagccgctg cccaccctcc cccaccctgg ccgggttctt
     1441 gcctggcttg tgcgtccttg tgacaattcc tgacacggat gcgcaccggg cagtggcacg
     1501 tccaggtgct gtgcgcgccc gggcccacaa agctcctttg gacgctcatg gcagcccctt
     1561 ggtggaggag gtcgggtgcc caccccctcc ccgcccactg cggaagccgg cgacccacgg
     1621 agctcgctct cggccgccgc cacccctctg tgttcgcgcc cttccgagct ctgatccgac
     1681 gctttgtttc ttctcagtgg gttcagggcc tgggccagcc tttacctacc tcccccaccc
     1741 aaaaccggca aaagctcaga gcaccttgtc tgccaaaaga cagggagctg ggatggtgcg
     1801 ggttggtctc taaaccggcg tggggaaaaa agaccctccg tacaaagccg cagggtgggg
     1861 ctgtcgcaag ggcggaaccg agagggtagc tgggggcggg gttcccaggg ccaagagggg
     1921 ccattgtcct ccctggagcc cggcgccccc acagccagct cctctgggag acagcccctc
     1981 ctttcgaatg cgcggggccc tcagaccgcg cccggcccag cgctggggga tccttggctg
     2041 cgggaggggc gccgcattgc gcgcggcggg cggggacgcg cggtgcggag cctgcgggcc
     2101 gggcggggct ctgcggcggc gcctcccgat tggccacccg cggtgacatc agccgatgcg
     2161 aagggcgggg ccgcggctat aagagcgcgc ggccgcggtc cccgaccctc agcagccagc
     2221 ccggcccgcc cgcgcccgtc cgcagccgcc cgccagacgc gcccagtggc gcgccgtcga
     2281 ctgcagaggc ctgcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     2341 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     2401 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     2461 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2521 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2581 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2641 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2701 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2761 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2821 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2881 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2941 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3001 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3061 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3121 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3181 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3241 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3301 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3361 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3421 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3481 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3541 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3601 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3661 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3721 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3781 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3841 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3901 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3961 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4021 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4081 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4141 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4201 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4261 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4321 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4381 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4441 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     4501 aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
