LOCUS       sequence-157015-        5771 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High copy number plasmid containing hs1218 MiniPromoter insert..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5771)
  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 5771)
  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: pEMS1957
FEATURES             Location/Qualifiers
     source          1..5771
                     /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"
     regulatory      1133..1142
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3550..3566)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3550..3566)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3563..3585)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3574..3590
                     /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(3598..3604,3605..3622,3623..3628))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3643..3664
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3781..3798)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3952..4540)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4032..4051)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4711..5502,5503..5571))
                     /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     5334..5353
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5572..5676)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5744..5762
                     /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 ctttcgtgga gaggactttc cagtcagtca gtcagtcgtc tcttggaacg gcactgcgct
      961 tggtttagtt gtcaagagag aataactctc ggacgcggaa ggattcgtag cgtatatcgc
     1021 ctattgcgga agcgtaacca agtcttacga ccccaaaaga ggttgactga actgggacgc
     1081 ggcactggcg cagatcacac tgtccgcgat aaaatcgggt gggtcatggc gcgccgccat
     1141 ggcgccgcgc tctgcttctg gaaggctgcg ctccgcggcg tggatgctcc ggggaaagtt
     1201 gcgctccgcg gcagggatgc tcctgggaag gttgcgctcc gcggcaggga tgctctgggg
     1261 aaggctggtc ctggccgagg atcgggaacg cgccgctcgc tctgcttctc ttgtcttcgc
     1321 ttgtctctgg atggaaccag atttggttct gagtagctgt cagcgtctgg tgacctgctc
     1381 gccgccctgc gcctttaagg agtcttcacc ggccccgccc actctccgct gggccaatca
     1441 gcgagccgga ggaggccttg gggccaggaa tcttccagca gtttcgcgtc tggtggagct
     1501 tccccgcctc ccttgagtaa tcggagttgt gggttccgcc cttgtccaga actctccaga
     1561 ggtttctggg gttcactgga gagtacggat tcctgagggg gagggtgtgg ggaagtgctg
     1621 gtgctactag tgacactgtt gctatggcga cgcattacta aggcctgtgt ggaatggaca
     1681 agaaagatca cctctagctc ggtgttgtgt acagtttgtt gtgatttgtg gggtttcgcc
     1741 aactcgcaca gttctgaata tgggggttaa aggctaaaac ttaagggcta aaacttctcc
     1801 ccgccaagtt taggagaccc agggagatgc ctgggggcgt gtccggtgac gtgatcctct
     1861 ccaatcgcgt tacaatggca gtgctgcctc tgacctcatg gactaattta ggaactagag
     1921 gctctgtccc agcacaggct caaagttgcc gcgtggggcg gggtgggggg tgggggggac
     1981 cccggctgct cagtttggat gttcctggag ctctcagaaa agatctactc cccagagtac
     2041 aaaggtaatg gctatactat aaagttgtca tttcaatgga gttttacttt ccacccaagg
     2101 gacagggcat tcatcatgaa catcctccat gccaagcaat cacagcatca ctgatcacct
     2161 tttgctcggg ccccatatat tgcaactatt ccagttgtgc catacctggc agagcagatt
     2221 ctgcagatcc taacctcttg ttgagactaa aataatttct taaacgcacg atagctacat
     2281 aactctccca gccattttct gaaccacgat ttttctcatt ttgcttcctt tgtgttccat
     2341 tcaaggtttt cctgcgctga gacaatcttt tacatttttt tcttcttctg cctgcatcaa
     2401 agttaagact ttgttaggga tgctttgcta tcggcatctg ttacctcttt ttcttagggg
     2461 ctaaggagaa ggctgctgtg tggctgaaaa agactctcat tcagtttgga attgcctccc
     2521 tttgctttcc tgtcacactg tcaagctcaa tcagaacgca atgaaagtga tacagaggcg
     2581 aggctttgct ggcgagactt agaacaaaga cagctccata gcctagtaca gatttcacat
     2641 ccaatttaac tctttggaaa aggatttcag gaaatacata gtcaagagtt gaaaaaatgg
     2701 agttgggtct actataaagg ctaaaagtat tatcctggct tagataagct acaaagatag
     2761 gttgatacta ttttctagca taaaccacat tagccggcat tacacattga gctttcttta
     2821 ctcgttaagt tgcaaggaga tttgagagga atttagcagc tacataaaat gactctttca
     2881 tgattctagc tttaccagtc attttagttg tacaaggtgc atgaatagca aggacctaaa
     2941 tcatgttgtt taacaaatct ggtattagag ggttagtctc agtttactta ttaagaattt
     3001 agttgagtaa aggattttag gactcagttc tttgtgtaga tgacaatgaa cggcccaata
     3061 cttgtatttt tgggggaaaa catacatcag ctacacatgt tttgaactgt gctgttggac
     3121 ccataactaa tatattgcac tgaatatatt tttctgcagg aactaactgt aaagaagaaa
     3181 cttcattgag agggacatag gtcaatggtg acttacttca gtggcctcta ctattacttc
     3241 ctttctgaag cctagtttct acttgttctc tgtctcacaa gagactcagt ttaattctct
     3301 gaatttttca aactgaggtc atgacaaact tggtttaatt ctctgagctt tcaaacttag
     3361 ttaatgattc catttggaag aatcatggcc ctcattttcc cttgatggat ttgaagcata
     3421 ctgaaaactg attcacaaag cacgtagaat cttgaacatt ctatacagca gcacgcgtag
     3481 gttgagacgc agtcagtcag tcagtgaaag aggaagggct ggaagaggaa ggagcttggc
     3541 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3601 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     3661 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     3721 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3781 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3841 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3901 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3961 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4021 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4081 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4141 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4201 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4261 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4321 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4381 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4441 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4501 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4561 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4621 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4681 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4741 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4801 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4861 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4921 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4981 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5041 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5101 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5161 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5221 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5281 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5341 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5401 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5461 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5521 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5581 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5641 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5701 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     5761 gccctttcgt c
//
