LOCUS       sequence-160449-        7019 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High copy number plasmid containing Ple305 MiniPromoter insert..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7019)
  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 7019)
  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: pEMS1964
FEATURES             Location/Qualifiers
     source          1..7019
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..144
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             2905..4095
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(3257..3276)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             4734..4925
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4919..4941)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5027..5167
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5082..5101
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5182..5199)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5353..5941)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5433..5452)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6112..6903,6904..6972))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6735..6754
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(6973..7019,1..58))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
       61 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
      121 ataaaaatag gcgtatcacg aggccctttc gtcttcaaga attccagtca gtcagtcagt
      181 gaagactgtg ctggcgcgcc gaggacgccc ttcaggaacg ccctggggtg ggccggtcgg
      241 caggccgccc ctccccgcgg acctgcgcgc ggggaaacga tggtggggcc ctggcccgcg
      301 ccctacccat ttatacccgg cgtgggaacc tgcggggcag cgggtggcgg ggcgcagcca
      361 atggaggcgc gaggccgggc gcagccgcgt gttcattggc tgcttgtgcc ggggccgcgc
      421 ggaggtgcgg ctggcaccgc tttaggaggt tattgtgagc gcctgtgcca gccgcgcggg
      481 acaggccccc tgagcggagt aagggcgcct tttcaaggca agcggggctg ttgaaggagg
      541 actggtcgca ctggtcaggc tgcaactcac gtgccttcct ccagcaactg tctactcaaa
      601 aacaaccttt ttcttctttt tcttttttgg tgacaatctg attataacta ctctgttctt
      661 attgacccta tgaggctttt ttttcacaat ttggaagaat taacagttat agcctccatg
      721 aataaatata acacttagct ccttcttact aactaaatgc aattggggaa taaatacagt
      781 gagtgttcac acacacacac acacacacac acacccacaa aagaaaaaaa aaagaaaaaa
      841 agaaaaaggt gaagaataga aaattgttta agtggccaaa gaaactttat cctattgtgt
      901 tgcaggccaa gctatgccag ctatatgtaa atgtaagcag tggcctgtgt tcctgtgtcc
      961 actttgaagt tgccatctga atttaattct gcctttgtga ttaaatcagg ccattacgtt
     1021 taatctcact taatagtaag aaacgtcctt tgttgttttc tataggcaac atattctgcc
     1081 cctaacatat acagcttggt attcacactt tactattaat ccccgatggc tgataattgc
     1141 ttcatctcca agccctctgt ccaattcaca aggggctgat tgcaactatt cttcccctat
     1201 acattaatgc actgtttggg tcttgcttct ttggctcagt gaaccctatc aagctgtcca
     1261 cattttctgg gcatgtgggt gtattgtgag atgttaaatt aaattccgtg ttgacctccc
     1321 taaacaccag aatctattag cccctgcaac gtagctgatt gacaagtgga actacttttc
     1381 cactcaaaat cctgcaataa gaaacaacaa caacaggctg aatattttgt ttctaggcag
     1441 agtctggggt cttcttattg ctgtcttgtt atttacatgt tcatgactac ggcaggacca
     1501 attgcattat tgccacgatg ttgcttcatt gatcttccat ttgtctgcct tatcatttct
     1561 gttggccaaa ggaatgacta ttggggacgg accccagcct cactggcagg gagagcctgg
     1621 cccccagcac cttgccgggg aggctcctct gggcctgact gagccccatt cagcccccat
     1681 gtgcagagga tgactcagcc tgtgacttca ttcacccagc ctggtgttta tccactcacc
     1741 acaaaccggt tgatactgtt tggagctacc ccgaggtgac gctctgctgg tggagaatga
     1801 catgcagtca gcttgtggcc tctggggcca aaacagctga aatgctgtgg cttctgaaga
     1861 acccattgtc tgctagtcaa gcgtgaggac aaggaaggtg gtgtctgaag agttggtttg
     1921 gggcagaaca aaaatatcct ctgagcctcc ctgaaggtag tagctgaagc caaatgtttc
     1981 cgaggcttac actgatacaa taaaaagaga caatttgctt ggaaggctgt ggctaattta
     2041 taacacaatg tccaatgtca tggttagttt tgttgcaaaa atttggcagt ttaacccaga
     2101 gtctcaggca gcctgaaaag atgcgagcgg ttctggccgc agcatctctg gcttgctgga
     2161 tgttccgaat ggttcaaaca agccctttca tagagacgca gggccaagtg cgctcaaagg
     2221 aatgagggaa acaaaggtgg ctctgaagtc atttccccgc actgggtcca gccatccggt
     2281 ttcactcctt ccccattgaa ccggcagaat gcaacaggaa gagcggggat gaaagagaac
     2341 aaacaggccg atgtgtttgg ggagagaatg gagagaggcc gcggcgctga gctgctaaac
     2401 tttttcattc taggttttca gaaaggagtg aatgatgaat tatagaaacc tgaaaggtaa
     2461 taaaatatgg gtcactgaag cgctgaggcc aagttcctaa aacacctgca gaggcagaag
     2521 gttctggaag tcagtgcagc cccacggttg cccgggcaga caccctgccc gagacacaga
     2581 aggggacttg ttcacaagct cacctgttgt tttcccgttg gctcccggtc cttccccgtc
     2641 tgtaagagct gacggctgga tctgactcca gattagacca ccaggggaag aaaggccgtg
     2701 tgaagacttc ccagggtgca gttaaggaga ccagagctcc aaggccaccg cctttttctc
     2761 tcggaggcgc tctgacctgg aaatattaac tcagtggccg gccaggttga gacgcagtca
     2821 gtcagtcagt gaaaggagat aggatccaag ctttaatgcg gtagtttatc acagttaaat
     2881 tgctaacgca gtcaggcacc gtgtatgaaa tctaacaatg cgctcatcgt catcctcggc
     2941 accgtcaccc tggatgctgt aggcataggc ttggttatgc cggtactgcc gggcctcttg
     3001 cgggatatcg tccattccga cagcatcgcc agtcactatg gcgtgctgct agcgctatat
     3061 gcgttgatgc aatttctatg cgcacccgtt ctcggagcac tgtccgaccg ctttggccgc
     3121 cgcccagtcc tgctcgcttc gctacttgga gccactatcg actacgcgat catggcgacc
     3181 acacccgtcc tgtggatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca
     3241 ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac
     3301 ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga
     3361 ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg cggcggtgct caacggcctc
     3421 aacctactac tgggctgctt cctaatgcag gagtcgcata agggagagcg tcgaccgatg
     3481 cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc
     3541 gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc
     3601 tgggtcattt tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt
     3661 gcggtattcg gaatcttgca cgccctcgct caagccttcg tcactggtcc cgccaccaaa
     3721 cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct gggctacgtc
     3781 ttgctggcgt tcgcgacgcg aggctggatg gccttcccca ttatgattct tctcgcttcc
     3841 ggcggcatcg ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga tgacgaccat
     3901 cagggacagc ttcaaggatc gctcgcggct cttaccagcc taacttcgat cactggaccg
     3961 ctgatcgtca cggcgattta tgccgcctcg gcgagcacat ggaacgggtt ggcatggatt
     4021 gtaggcgccg ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg
     4081 gccacctcga cctgaatgga agccggcggc acctcgctaa cggattcacc actccaagaa
     4141 ttggagccaa tcaattcttg cggagaactg tgaatgcgca aaccaaccct tggcagaaca
     4201 tatccatcgc gtccgccatc tccagcagcc gcacgcggcg catctcgggc agcgttgggt
     4261 cctggccacg ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta ggctggcggg
     4321 gttgccttac tggttagcag aatgaatcac cgatacgcga gcgaacgtga agcgactgct
     4381 gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt
     4441 aaagtctgga aacgcggaag tcagcgccct gcaccattat gttccggatc tgcatcgcag
     4501 gatgctgctg gctaccctgt ggaacaccta catctgtatt aacgaagcgc tggcattgac
     4561 cctgagtgat ttttctctgg tcccgccgca tccataccgc cagttgttta ccctcacaac
     4621 gttccagtaa ccgggcatgt tcatcatcag taacccgtat cgtgagcatc ctctctcgtt
     4681 tcatcggtat cattaccccc atgaacagaa atccccctta cacggaggca tcagtgacca
     4741 aacaggaaaa aaccgccctt aacatggccc gctttatcag aagccagaca ttaacgcttc
     4801 tggagaaact caacgagctg gacgcggatg aacaggcaga catctgtgaa tcgcttcacg
     4861 accacgctga tgagctttac cgcagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc
     4921 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     4981 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc
     5041 agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt
     5101 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     5161 catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5221 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5281 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5341 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5401 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5461 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5521 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5581 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5641 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5701 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5761 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     5821 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5881 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5941 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6001 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6061 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6121 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6181 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6241 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6301 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6361 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6421 cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6481 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6541 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6601 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6661 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6721 ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6781 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6841 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6901 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6961 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtc
//
