LOCUS       sequence-435068-        3295 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 3295)
  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 3295)
  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-WPRE-G
FEATURES             Location/Qualifiers
     source          1..3295
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     13..30
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    147..168
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(183..188,189..206,207..213)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    221..237
                     /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        251..269
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     251..268
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    306..894
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(359..379)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(777..788)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(920..939)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(921..939)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(949..971)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1165..1184)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1284..1306
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1344..1362)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1430..1534
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1535..1603,1604..2395)
                     /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(1753..1772)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2566..3154
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3055..3074
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
       61 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
      121 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
      181 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
      241 cacacgagct atttaggtga cactatagaa gtgaagcttg gtctcaacta taatcgaatt
      301 ccgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
      361 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
      421 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
      481 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
      541 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
      601 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
      661 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
      721 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
      781 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
      841 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcggg
      901 tattgagacc gaattctcgc cctatagtga gtcgtattac ctggccgtcg ttttacaacg
      961 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     1021 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     1081 cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     1141 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     1201 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     1261 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     1321 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     1381 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     1441 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     1501 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     1561 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     1621 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     1681 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     1741 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     1801 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     1861 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     1921 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     1981 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     2041 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     2101 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     2161 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     2221 tgctgataaa tctggagccg gtgagcgtgg ctctcgaggt atcattgcag cactggggcc
     2281 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     2341 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     2401 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     2461 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     2521 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     2581 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     2641 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     2701 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     2761 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     2821 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     2881 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     2941 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3001 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     3061 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     3121 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     3181 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     3241 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccga
//
