LOCUS       sequence-126156-        5251 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides the S. Lycopersicum fruit promoter E8 as a level 0 
            GoldenBraid part.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5251)
  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 5251)
  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: pPromE8  (GB0914)
FEATURES             Location/Qualifiers
     source          1..5251
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(2301..2319)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2302..2319)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2337..2353)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2337..2353)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2350..2372)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2361..2377
                     /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(2385..2391,2392..2409,2410..2415))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2430..2451
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2568..2585)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2739..3327)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2819..2838)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3498..4289,4290..4358))
                     /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     4121..4140
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4359..4463)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4541..4996)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4678..4699)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4890..4909
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5122..5144
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5136..5153
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5137..5153
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5160..5179
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5160..5178
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      5209..5218
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      5213..5222
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ctcgggagtc cctaatgata ttgttcacgt aattaagttt tgtggaagtg agagagtcca
       61 attttgataa gaaaagagtc agaaaacgta atattttaaa agtctaaatc tttctacaaa
      121 taagagcaaa tttatttatt ttttaatcca ataaatatta atggaggaca aattcaattc
      181 acttggttgt aaaataaact taaaccaata accaaagaac taataaatcc tgaagtggaa
      241 ttattaagga taaatgtaca tagacaatga agaaataata ggttcgatga attaataata
      301 attaaggatg ttacaatcat catgtgccaa gtatatacac aatattctat gggatttata
      361 atttcgttac ttcacttaac ttttgcgtaa ataaaacgaa ttatctgata ttttataata
      421 aaacagttat aagaaccatc atttttaaca acatagatat attatttcta atagtttaat
      481 gatactttta aatcttttaa attttatgtt tcttttagaa aataaaaatt caaaaaatta
      541 aatatattta caaaaactac aatcaaacac aacttcatat attaaaagca aaatatattt
      601 tgaaaatttc aagtgtccta acaaataaga caagaggaaa atgtacgatg agagacataa
      661 agagaactaa taattgagga gtcctataat atataataaa gtttattagt aaacttaatt
      721 attaaggact cctaaaatat atgataggag aaaatgaatg gtgagagata ttggaaaact
      781 taataattaa ggattttaaa atatatggta aaagataggc aaagtatcca ttatcccctt
      841 ttaacttgaa gtctactagg cgcatgtgaa agttgatttt ttgtcacgtc atatagctat
      901 aacgtaaaaa aagaaagtaa aatttttaat tttttttaat atatgacata ttttaaacga
      961 aatataggac aaaatgtaaa tgaatagtaa aggaaacaaa gattaatact tactttgtaa
     1021 gaatttaaga taaatttaaa atttaataga tcaactttac gtctagaaag acccatatct
     1081 agaaggaatt tcacgaaatc ggcccttatt cgaaaataac ttttaaataa tgaattttaa
     1141 attttaagaa ataatatcca atgaataaat gacatgtagc attttaccta aatatttcaa
     1201 ctattttaat ccaatattaa tttgttttat tcccaacaat agaaagtctt gtgcagacat
     1261 ttaatctgac ttttccagta ctaaatatta attttctgaa gattttcggg tttagtccac
     1321 aagttttagt gagaagtttt gctcaaattt taggtgagaa ggtttgatat ttatcttttg
     1381 ttaaattaat ttatctaggt gactattatt tatttaagta gaaattcata tcattacttt
     1441 tgccaacttg tagtcataat aggagtaggt gtatatgatg aaggaataaa caagttcagt
     1501 gaagtgatta aaataaaata taatttaggt gtacatcaaa taaaaacctt aaagtttaga
     1561 aaggcaccga ataattttgc atagaagata ttagtaaatt tataaaaata aaagaaatgt
     1621 agttgtcaag ttgtcttctt ttttttggat aaaaatagca gttggcttat gtcattcttt
     1681 tacaacctcc atgccacttg tccaattgtt gacacttaac taattagttt gattcatgta
     1741 tgaatactaa ataatttttt aggactgact caaatatttt tatattatca tagtaatatt
     1801 tatctaattt ttaggaccac ttattactaa ataataaatt aactactact atattattgt
     1861 tgtgaaacaa caacgttttg gttgttatga tgaaacgtac actatatcag tatgaaaaat
     1921 tcaaaacgat tagtataaat tatattgaaa atttgatatt tttctattct taatcagacg
     1981 tattgggttt catattttaa aaagggacta aacttagaag agaagtttgt ttgaaactac
     2041 ttttgtctct ttcttgttcc catttctctc ttagatttca aaaagtgaac tactttatct
     2101 ccttctttgt tcacatttta ttttattcta ttataaatat ggcatcctca tattgagatt
     2161 tttagaaatt attctaatca ttcacagtgc aaaagaagaa tgcgagacca ctcatcgccc
     2221 aatcactagt gaattcgcgg ccgcctgcag gtcgaccata tgggagagct cccaacgcgt
     2281 tggatgcata gcttgagtat tctatagtgt cacctaaata gcttggcgta atcatggtca
     2341 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     2401 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     2461 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     2521 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     2581 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2641 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2701 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2761 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2821 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2881 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2941 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3001 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3061 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3121 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3181 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3241 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3301 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3361 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3421 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3481 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3541 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3601 ggcttaccat ctggccccag tgctgcaatg ataccgcgcg acccacgctc accggctcca
     3661 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3721 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3781 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     3841 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     3901 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     3961 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4021 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4081 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     4141 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4201 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4261 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4321 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4381 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4441 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga tgcggtgtga
     4501 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag cgttaatatt
     4561 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     4621 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     4681 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     4741 gtctatcagg gcgctggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     4801 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     4861 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     4921 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     4981 ccgctacagg gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     5041 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     5101 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattgt
     5161 aatacgactc actatagggc gaattgggcc cgacgtcgca tgctcccggc cgccatggcg
     5221 gccgcgggaa ttcgattggg cgatgagtgg t
//
