LOCUS       sequence-157016-        3805 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High copy number plasmid containing PR2/3:Con MiniPromoter insert..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3805)
  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 3805)
  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: pEMS1958
FEATURES             Location/Qualifiers
     source          1..3805
                     /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"
     CDS             695..706
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(1584..1600)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1584..1600)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1597..1619)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1608..1624
                     /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(1632..1638,1639..1656,1657..1662))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1677..1698
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1815..1832)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1986..2574)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2066..2085)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2745..3536,3537..3605))
                     /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     3368..3387
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3606..3710)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3778..3796
                     /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 accaaatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcatcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgcaac gcgatgacga tggatagcga
      421 ttcatcgatg agctgacccg atcgccgccg ccggagggtt gcgtttgaga cgggcgacag
      481 atcgacactg ctcgatccgc tcgcaccggc cggcctctac ttgtgggaga ggacaggagg
      541 gagaggtgcc gtggtgagac tgaccctcgg gcccacgggt ggatgatggc aaaggtcctg
      601 tggcaggtgt gggagaagca gggctgcggc cagctgccct gagaggcttc aaattcaggc
      661 cagatctgct ctgggcaaga ggaagctgtg ggtttgggag cttcagagac aggggagcag
      721 gcctgtcact caccctcttt tccttaaagg ctgtcacccc cagcagtgca ccctgcagcc
      781 tgcctatccc ctgtgcagct ccagctccgt ctgtccccag caatgcaccc cacgcccatg
      841 tcccccactc cctatgatgc tctcgcgcct tctggagcag cagtggtacc agctggaggc
      901 tgtggaaaaa ttgagaggag acaaaagtgg gaagagcagg ccgtggagag agtgtctaaa
      961 cacagggact atttggagat tgtcttattt ttggtttgaa atgggcccac ctgctagtat
     1021 tgtacaaacg gctctgctcg ctaggccagg cctgcagtgg ctacgggcct atccttcact
     1081 ggtcagcctg gagcagctcc tcggacctca gtctgctcat ccgtgggtag aggcgacgca
     1141 gaccccatcc cactggggac tgggccagac agcctgtgag gcagtcagct ggtgcctggc
     1201 cagagggtgt ctgaaaggcc tcttggctgc agggtgcatc tgcttttggg acagctcttc
     1261 agagccatct cagaagggac agcatccgcc tgtctacttc aactcccact gatgacgtcc
     1321 atgtgtcatt agtgccaatt agaggagggc agcaggctga gtgcttggcc tggggcgcaa
     1381 gcttgtggga gggataattg gattacccgc tagactaatg tgattacccg tgctgcctgg
     1441 acccacccca ttcaggacca gggcataaat ggccaggtgg gaccagagag ctcaccccag
     1501 ccggcgcgcc ggatcgacga gagcagcgcg actggatcag ttctggacca gcgagctgtg
     1561 ctgcgactcg tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     1621 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     1681 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     1741 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     1801 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     1861 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     1921 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     1981 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2041 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2101 gtgcgctctc ctgttccgac cctgtcgctt accggatacc tgtccgcctt tctcccttcg
     2161 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2221 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2281 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2341 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2401 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2461 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2521 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2581 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     2641 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2701 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     2761 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     2821 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     2881 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     2941 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3001 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3061 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3121 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3181 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3241 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3301 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3361 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3421 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3481 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     3541 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     3601 ctcatactct acctttttca atattattga agcatttatc agggttattg tctcatgagc
     3661 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     3721 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     3781 aggcgtatca cgaggccctt tcgtc
//
