LOCUS       sequence-435069-        2913 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  golden gate cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2913)
  AUTHORS   Delhaye L, Moschonas GD, Fijalkowska D, Verhee A, De Sutter D, Van 
            de Steene T, De Meyer M, Grzesik H, Van Moortel L, De Bosscher K, 
            Jacobs T, Eyckerman S
  TITLE     Leveraging a self-cleaving peptide for tailored control in proximity
            labeling proteomics.
  JOURNAL   Cell Rep Methods. 2024 Jul 15;4(7):100818. doi: 
            10.1016/j.crmeth.2024.100818. Epub 2024 Jul 9.
  PUBMED    38986614
REFERENCE   2  (bases 1 to 2913)
  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: "Cell Rep 
            Methods."; date: "2024-07-15"; volume: "4(7):100818. doi"; pages: " 
            10.1016/j.crmeth.2024.100818. Epub 2024 Jul 9"
COMMENT     Sequence Label: pGGF-bGH polyA-G
FEATURES             Location/Qualifiers
     source          1..2913
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     205..222
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    339..360
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(375..380,381..398,399..405)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    413..429
                     /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        443..461
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     443..460
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     polyA_signal    486..710
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(730..749)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(731..749)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(759..781)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(975..994)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1094..1116
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1154..1172)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1240..1344
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1345..1413,1414..2205)
                     /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     complement(1563..1582)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(2376..2913,1..51)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2865..2884
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
       61 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      121 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
      181 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
      241 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
      301 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
      361 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
      421 ggataacaat ttcacacgag ctatttaggt gacactatag aagtgaagct tggtctcaac
      481 taagcctgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
      541 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
      601 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
      661 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg tattgagacc
      721 gaattctcgc cctatagtga gtcgtattac ctggccgtcg ttttacaacg tcgtgactgg
      781 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
      841 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
      901 gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
      961 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     1021 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     1081 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     1141 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     1201 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     1261 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     1321 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     1381 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     1441 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     1501 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     1561 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     1621 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     1681 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     1741 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     1801 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     1861 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     1921 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     1981 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     2041 tctggagccg gtgagcgtgg ctctcgaggt atcattgcag cactggggcc agatggtaag
     2101 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     2161 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     2221 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     2281 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     2341 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     2401 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     2461 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     2521 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     2581 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     2641 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     2701 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     2761 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     2821 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     2881 ctttatagtc ctgtcgggtt tcgccacctc tga
//
