LOCUS       sequence-127520-        3460 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides the 4xLexA Operator fused to CaMV 35s minimal promoter as a
            level 0 GoldenBraid part.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3460)
  AUTHORS   Sarrion-Perdigones A, Vazquez-Vilar M, Palaci J, Castelijns B, 
            Forment J, Ziarsolo P, Blanca J, Granell A, Orzaez D
  TITLE     GoldenBraid 2.0: a comprehensive DNA assembly framework for plant 
            synthetic biology.
  JOURNAL   Plant Physiol. 2013 Jul;162(3):1618-31.
  PUBMED    23669743
REFERENCE   2  (bases 1 to 3460)
  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: "Plant 
            Physiol."; date: "2013-07"; volume: "162(3)"; pages: "1618-31"
COMMENT     Sequence Label: pOpLexA::mini35s (GB0733)
FEATURES             Location/Qualifiers
     source          1..3460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(82..128)
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     promoter        complement(510..528)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(511..528)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(546..562)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(546..562)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(559..581)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    570..586
                     /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(594..600,601..618,619..624))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    639..660
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(777..794)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(948..1536)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1028..1047)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1707..2498,2499..2567))
                     /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     2330..2349
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2568..2672)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2750..3205)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2887..2908)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3099..3118
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3331..3353
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3345..3362
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3346..3362
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3369..3388
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3369..3387
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3418..3427
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3422..3431
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ctcgcatttc gactagaata gtaaattgta atgttgtttg ttgtttgttt tgttgtggta
       61 attgttgtaa aaataccgga gtcctctcca aatgaaatga acttccttat atagaggaag
      121 ggtcttgcga aggggagcat attacatggc attaccacca tatacatatc catatacata
      181 tccatatcta atcttacctc gactgctgta tataaaacca gtggttatat gtacagtact
      241 gctgtatata aaaccagtgg ttatatgtac agtacgtcga ctgctgtata taaaaccagt
      301 ggttatatgt acagtactgc tgtatataaa accagtggtt atatgtacag tacgtcgagg
      361 ggatgataat gcgattagtt ttttagcctc gacctgcagc ccaagctgtc acgagaccac
      421 tcatcgccca atcactagtg aattcgcggc cgcctgcagg tcgaccatat gggagagctc
      481 ccaacgcgtt ggatgcatag cttgagtatt ctatagtgtc acctaaatag cttggcgtaa
      541 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
      601 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
      661 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
      721 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
      781 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
      841 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
      901 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
      961 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     1021 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     1081 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     1141 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     1201 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     1261 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     1321 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     1381 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     1441 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     1501 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     1561 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     1621 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     1681 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     1741 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     1801 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgcga cccacgctca
     1861 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     1921 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     1981 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     2041 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     2101 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     2161 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     2221 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     2281 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     2341 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     2401 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     2461 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     2521 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     2581 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     2641 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgat
     2701 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga aattgtaagc
     2761 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     2821 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     2881 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     2941 cgaaaaaccg tctatcaggg cgctggccca ctacgtgaac catcacccta atcaagtttt
     3001 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     3061 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     3121 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     3181 cttaatgcgc cgctacaggg cgcgtccatt cgccattcag gctgcgcaac tgttgggaag
     3241 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     3301 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     3361 gtgaattgta atacgactca ctatagggcg aattgggccc gacgtcgcat gctcccggcc
     3421 gccatggcgg ccgcgggaat tcgattgggc gatgagtggt
//
