LOCUS       sequence-157014-        5548 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High copy number plasmid containing hs671 MiniPromoter insert..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5548)
  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 5548)
  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: pEMS1953
FEATURES             Location/Qualifiers
     source          1..5548
                     /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"
     CDS             complement(999..1016)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      3061..3070
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3327..3343)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3327..3343)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3340..3362)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3351..3367
                     /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(3375..3381,3382..3399,3400..3405))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3420..3441
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3558..3575)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3729..4317)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3809..3828)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4488..5279,5280..5348))
                     /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     5111..5130
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5349..5453)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5521..5539
                     /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 ctacgcgtgc tgagattgtg tgatgtggag
      961 aaaaggaaga tatgcagaga tgttagcatg gccacagggt gatggtggtg gtggtgctgg
     1021 ggcaagtgtg cactactaat aggagaagat gtcaaaaaaa atgaagcata ataatagtgc
     1081 aatttatcct tgctaaacgt actgcagtgt gctataatta aatatgaaaa cctaaattta
     1141 tctctatgtt cggttgtcat taatttaaca gggagtgcac atgttctaat ctaatggttg
     1201 attttttttc tccatgtgca gcatattaaa atgtatatta gcctcaaggt gcataaacac
     1261 aaaataatag accaaaggtt ttggcctgtg gagaaaaaaa atccatgctc aaaggccgtc
     1321 aagtagctgt gatgcagggg agacactcac ttagcctcat taagccaaag ctctcttttg
     1381 cttgtctttt gttctgcatg acagcagcct ctttgttgta taataattca ttccaagttc
     1441 gttccaagct accgaaactg gtgaactgtg gccattttaa tgagaagtat tggtggtatg
     1501 cataatctgg aagccacgtg ggccagacaa tcacagagca ctgcagccag caaaatgaga
     1561 tattgtcgaa atcacaaatt tgtcaagttg tttttggaca gggtattata ataagcacta
     1621 aaatatgcat cctattactc tgcatctgcc agtcaaagat cactgtaatg aatgcgtgct
     1681 gcagggtata accaggccgt tttcctaagt aaaacaaacc acagagtaca gaaaagaatt
     1741 gtgtgggttt tttttttttt ttttggcatt gtttgacttt tttttttttc ctaatccttt
     1801 tgcaccagga gtcaggttct cctggcatgg tgtccactgc tagcagtgaa tacacaggcc
     1861 ctgctggtgc cagggagtta acatgaatac attgcttttt taataggtgt gcttaccatc
     1921 tgagaataca aaacctcaaa ctcaaggctt caattgagtc tactgaagaa ttcctgggag
     1981 agaaaaggga aaaaggagga gaaaaaaata tctctcccaa tgatcctaag ccttatgttt
     2041 tatccttttc tacttcaaat ggcttagcct ttgggacttg ttttgttgtt tgtttttctg
     2101 tttgtttatt tttgcttttt tgtctgtctg ttgatttatt tgctttttag atgaacctaa
     2161 agctttggga agaaatgggt gagatgctag agctccagga acatccaaac tgagcagccg
     2221 gggtcccccc caccccccac cccgccccac gcggcaactt tgagcctgtg ctgggacaga
     2281 gcctctagtt cctaaattag tccatgaggt cagaggcagc actgccattg taacgcgatt
     2341 ggagaggatc acgtcaccgg acacgccccc aggcatctcc ctgggtctcc taaacttggc
     2401 ggggagaagt tttagccctt aagttttagc ctttaacccc catattcaga actgtgcgag
     2461 ttggcgaaac cccacaaatc acaacaaact gtacacaaca ccgagctaga ggtgatcttt
     2521 cttgtccatt ccacacaggc cttagtaatg cgtcgccata gcaacagtgt cactagtagc
     2581 accagcactt ccccacaccc tccccctcag gaatccgtac tctccagtga accccagaaa
     2641 cctctggaga gttctggaca agggcggaac ccacaactcc gattactcaa gggaggcggg
     2701 gaagctccac cagacgcgaa actgctggaa gattcctggc cccaaggcct cctccggctc
     2761 gctgattggc ccagcggaga gtgggcgggg ccggtgaaga ctccttaaag gcgcagggcg
     2821 gcgagcaggt caccagacgc tgacagctac tcagaaccaa atctggttcc atccagagac
     2881 aagcgaagac aagagaagca gagcgagcgg cgcgttcccg atcctcggcc aggaccagcc
     2941 ttccccagag catccctgcc gcggagcgca accttcccag gagcatccct gccgcggagc
     3001 gcaactttcc ccggagcatc cacgccgcgg agcgcagcct tccagaagca gagcgcggcg
     3061 ccatggcggc gcgccatgac ccacccgatt ttatcgcgga cagtgtgatc tgcgccagtg
     3121 ccgcgtccca gttcagtcaa cctcttttgg ggtcgtaaga cttggttacg cttccgcaat
     3181 aggcgatata cgctacgaat ccttccgcgt ccgagagtta ttctctcttg acaactaaac
     3241 caagcgcagt gccgtagcaa gagacgactg actgactgac tggaaagagg aagggctgga
     3301 agaggaagga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     3361 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     3421 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     3481 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     3541 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3601 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3661 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3721 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3781 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3841 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3901 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3961 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4021 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4081 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4141 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     4201 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4261 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4321 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4381 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4441 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4501 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4561 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4621 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4681 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4741 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4801 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     4861 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     4921 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     4981 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5041 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5101 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5161 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5221 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5281 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5341 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5401 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5461 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     5521 aataggcgta tcacgaggcc ctttcgtc
//
