LOCUS       sequence-157018-        5600 bp    DNA     circular SYN 15-APR-2025
DEFINITION  High copy number plasmid containing Ple302 MiniPromoter insert..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5600)
  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 5600)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pEMS1961
FEATURES             Location/Qualifiers
     source          1..5600
                     /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(3379..3395)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3379..3395)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3392..3414)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3403..3419
                     /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(3427..3433,3434..3451,3452..3457))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3472..3493
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3610..3627)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3781..4369)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3861..3880)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4540..5331,5332..5400))
                     /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     5163..5182
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5401..5505)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5573..5591
                     /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 gatgtccttt ttacttctat
      961 cttccataat attctaaaat tattttccct tgttttgttc ctatcctact tcctcttagt
     1021 ctactttgtt gacttcatta aaaaacaaaa aaccagttgt tggatacttg agctaaactg
     1081 ccttaaagaa tctgcagatt ttattttatt ttttttctct caagagggta aaaggaagag
     1141 agctacaatt tctaagaagc ctggcttggc tgtctgagtc tggcccccag gcagattagg
     1201 ccaaggtttt ggccaagtga aattgccaat tttctaaaag aaagggctag cacattgctc
     1261 attagagcat tctgattttg tctgcgcaat ctttctgcta ccccgcaatt tcctgttggt
     1321 tataaatgaa acctttctag ctgttaatgc agcctgtgaa tttttttaaa agcatgtaat
     1381 taatcatagg aggttggggg gattcactaa gcctgagtta catgggagaa gctggacaag
     1441 gcactaggac ctagaaggca tctatccacc ctggcaggaa tttcttgctt ggagctcaga
     1501 caacaaaggc atagagagat tggttttctt tctctcagca tctccaccca accagcagaa
     1561 aaccggtgag tggggctttt aagtgatttt caagaagaat gtaacagatg tcaaacggga
     1621 aaagcacaag gcaaagcctg ctctctctgt ctctctgtct cctcttctcc ttttttgcct
     1681 tattctatcc gattttttcc ctaagcttct acctgggatt ttcctttgga aaagtgagtt
     1741 tgatgttcct ttgttttcac tgtgatgtta atttagaata atactacctc tgatcctaaa
     1801 gcaaagcaaa gccttactgg catgcctggg gaaatgtttg ctgcttgcct tgaggaggtg
     1861 gggtctctta ccactgcagg ttgtctgaca gagacaatgc tgagctcagc ataggtcatg
     1921 gtgacattgg aaaaaaggcg gaattgagcc tggcagaccc attaggcacc agtctttctt
     1981 atctcctgtc ctcctggtcc cttgcaaata tattgatgtg gcagtgtgta gcagctgagc
     2041 cctgcttgct ttgtgagtcc ttttatcccc atctgtgaga tgcatgttaa tagtttggct
     2101 cgtaggatgt cactacattt gctagcattt gtggcttcag ttgtattggg tttcatgttt
     2161 tgattgtttg gggttcttgg tgggggaggg ggttcaacag aagggagaaa agcaaagcct
     2221 gacaaatgac catcttttct cagctaatgc acctgggcaa tatacaagtt tggggtgaat
     2281 tgcctgctgt gagggtaaat gtcacttcaa ttaaggtaga aacccagaac aatgaaaggt
     2341 gtgcttcctt ctaaaggtcc cgtatgctgt tcggagagtc atttgtgaat ctttcaacaa
     2401 ttaaattatt ccattaagag gtgttgctgc atcagtgggg agggggtgga gcacctgggg
     2461 gggaaaaaaa aggattttgt gaacaaatgg aaccggggga agacagagct agtaacttgt
     2521 taaataactt atttttctaa tcctttttcc ccccagctta tttcttatga atgtcggata
     2581 gctgcaccag cttggtgggg aaagggtttg atgaatagca caaagacact ggctgttccc
     2641 tggaggctgt ccctttaaag gagaatctta gtttattctg gggggagggg atgcacacat
     2701 tagagtagga aagagggctt ggaataaaat gaaaacactc ccccttcata gtcattgtac
     2761 tgaaatgcaa agactgcttc ctaagctgga gatgctaacc ttgggtagct ccttctgttc
     2821 tcttcaaggg gaattttgtc aggctatgga ttcatttaca actgttagtc atgtgggcat
     2881 gtgtgaggaa acagatgcca gttttaatgt atttagcccg aagttccaat ttgataggag
     2941 ccactgtcag taagtctcag gattttcagc tatttcaaaa tctccccttc tcctctgtct
     3001 ggaacagtgc caagagtgcc tccctctcta tctcttactc ccaaccccca caaccaccag
     3061 cacccccgcc cagcccctcc ttcttctcta ttaagatcaa tattcctgca ggtcaggggc
     3121 aagcagcaga tgggtcacag gcttttttca accagttctt ttcacaagca gcagattgca
     3181 gatctggatc tggctaatat ttaaaatccc ttcttttttc cttctccttg tccctttttg
     3241 tttttgcctc tcttcacccc catccctttc tcccacgctc aggtctctga ggttccaccg
     3301 gcgcgccacg aagagacgac tgactgactg actggaaaga ggaagggctg gaagaggaag
     3361 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     3421 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     3481 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     3541 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3601 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3661 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3721 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3781 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3841 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3901 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3961 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4021 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4081 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4141 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4201 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     4261 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4321 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4381 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4441 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4501 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4561 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4621 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4681 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4741 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4801 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4861 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4921 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4981 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5041 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5101 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5161 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5221 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5281 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5341 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5401 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5461 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5521 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     5581 tatcacgagg ccctttcgtc
//
