LOCUS       sequence-445481-        4359 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High Copy number plasmid containing Ple388 (PITX3 MiniPromoter) 
            insert.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4359)
  AUTHORS   Fornes O, Av-Shalom TV, Korecki AJ, Farkas RA, Arenillas DJ, 
            Mathelier A, Simpson EM, Wasserman WW
  TITLE     OnTarget: in silico design of MiniPromoters for targeted delivery of
            expression.
  JOURNAL   Nucleic Acids Res. 2023 Jul 5;51(W1):W379-W386. doi: 
            10.1093/nar/gkad375.
  PUBMED    37166953
REFERENCE   2  (bases 1 to 4359)
  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: "Nucleic 
            Acids Res."; date: "2023-07-5"; volume: "51(W1):W379-W386. doi"; 
            pages: " 10.1093/nar/gkad375"
COMMENT     Sequence Label: pEMS2371
FEATURES             Location/Qualifiers
     source          1..4359
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(23..39)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(23..39)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(36..58)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    47..63
                     /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(71..77,78..95,96..101))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    116..137
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(254..271)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(425..1013)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(505..524)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1184..1975,1976..2044))
                     /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     1807..1826
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2045..2149)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2217..2235
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2273..2295)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2395..2414
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2608..2630
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2622..2639
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2623..2639
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
       61 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
      121 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
      181 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
      241 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
      301 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
      361 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
      421 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
      481 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
      541 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
      601 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
      661 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
      721 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
      781 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
      841 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
      901 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
      961 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     1021 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     1081 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     1141 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     1201 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     1261 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     1321 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     1381 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     1441 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     1501 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     1561 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     1621 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     1681 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     1741 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     1801 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     1861 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     1921 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     1981 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     2041 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     2101 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     2161 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     2221 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     2281 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     2341 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     2401 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     2461 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
     2521 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     2581 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     2641 aattcggccg gcctaagcgc gcccccttcc gggggtgcag gacataacag cttcatcccc
     2701 agtgcgggtg ggaccccggc tgggcagccc cggtgggagc cagcgagtgg cttaggaggt
     2761 ccccgagccg gagaatatgc gctggcttgg gcgctctgtg acctgccccc accctggccc
     2821 ccagtcagtc ccaactccct agcttcctga cctccagcag tttgggggag cttaatcttc
     2881 agccagcctg tctccagctc agacaacagc ttggcgcccc cagcctttgc ctgaccagcc
     2941 cggggttaaa gttgaggcag caaatttatc cccagagata tatgtctagc acaaagctga
     3001 gattcacaag attctccctg agttttcctt acctctaaat gctggaataa ctgatgctca
     3061 ctgaattccg atatttgtat gaataaaaat gaagggattt agcaaatctt gttcctgcac
     3121 acagttgtca aaaaaaatct gacctttaca acaggaaaaa gaggggggat ttaaagaatg
     3181 tgagttccac aaacgggcaa gtaattgctc aataagcgtg gaacaaggaa aggtcagttg
     3241 tgatctgtgc acatgtggca gcccttcaca actcgaccta cagtggtaac agtggtaaca
     3301 gaatgctgat gttacagaag agacatcgga ttctttgggg atagagaaaa tacataatat
     3361 atattactat aaatgcaaaa ctgtactaaa agctaactca aatgtcaaag attacagttt
     3421 gaataaattt aaatataaaa gctagccgga gcgcggtctc tgccacgtgg ggaggggcgc
     3481 ggccgagttg ctgaagagcg cttctgattg gccagagggc ggggttcttg gcgtctcgcc
     3541 ggccagaccc ctccctcaaa ggcggggcct ggagatccac agctggaaag ggcggagccc
     3601 cagcagggca gctggaaagg ggcggggcct gacgcgcgcg gctcgccgcg gcgggctggg
     3661 ggcgccctgg tctgccataa agtgaatggg cgccggctgg gggtggcagt acgcggtgag
     3721 gctcactccc tccgagagtc caggagcgcc cgagcggaga ggcggcccgg gagcaggggg
     3781 gcggccccca ctccggccgg gtgcccggcc cctggcccct gcctgccctc tagatcgccg
     3841 ccgcagccgc cgctactggg agtctgcctg ttgcaggacg cactgtgagt gtttgaccat
     3901 tctaaggaca ggggctcatg gggccgagga gctattgctg gaccttggaa tccctaaaac
     3961 gagacctggg gcagctgaac tccggtgcct tgggaacagg tcgagaatgg gagggcggac
     4021 tgcagacctg gggccaggac ttgggaaaag ggaaactcag ggtaagaacc aggggatagg
     4081 gaagggggag actcagagga tccggaacga agccggagac ttgaaaccag gataggggct
     4141 tcgcgaggca gggagtaggg agtctggaaa ccattaccag aaagcgccct ggaatcccag
     4201 ggcacggggc aggcggccag ggagcgaaag gggttacaga ggccctggcc agcgggggag
     4261 ctggcggggc ggcccccagt gtgggggcag ttgacggcgc ccgatgcccc tcggcctagg
     4321 ctcagactcc tggggctggg agccggggtg cgcctggcg
//
