LOCUS       sequence-319733-        3279 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Golden Gate assembly of a pET28-compatible fusion-protein ORF 
            insert.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3279)
  AUTHORS   Haynes KA, Priode JH
  TITLE     Rapid Single-Pot Assembly of Modular Chromatin Proteins for 
            Epigenetic Engineering.
  JOURNAL   Methods Mol Biol. 2023;2599:191-214. doi: 
            10.1007/978-1-0716-2847-8_14.
  PUBMED    36427151
REFERENCE   2  (bases 1 to 3279)
  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: "Methods Mol
            Biol."; date: "2023"; volume: "2599:191-214. doi"; pages: " 
            10.1007/978-1-0716-2847-8_14"
COMMENT     Sequence Label: GGDest2-Amp
FEATURES             Location/Qualifiers
     source          1..3279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     8..30
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     22..39
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     23..39
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(226..242)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(226..242)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(239..261)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    250..266
                     /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(274..280,281..298,299..304))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    319..340
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(457..474)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(628..1216)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(708..727)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1387..2178,2179..2247))
                     /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     2010..2029
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2248..2352)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2420..2438
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2476..2498)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2598..2617
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2634..3089)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2771..2792)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2983..3002
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattggaggc tacagtcagt
       61 ggagaggact ttcactgact gactgactgc gtctcaacct gaattcggat ccaaggaggg
      121 gtcttcgaga agacctacta ggaagcttct gcagagcaag agacgactga ctgactgact
      181 ggaaagagga agggctggaa gaggaaggag cttggcgtaa tcatggtcat agctgtttcc
      241 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
      301 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
      361 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
      421 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
      481 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
      541 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
      601 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
      661 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
      721 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
      781 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
      841 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
      901 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
      961 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     1021 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     1081 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     1141 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     1201 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     1261 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     1321 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     1381 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     1441 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     1501 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     1561 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     1621 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     1681 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     1741 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     1801 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     1861 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     1921 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     1981 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     2041 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     2101 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     2161 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     2221 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     2281 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     2341 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     2401 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     2461 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     2521 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     2581 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc ataaaattgt
     2641 aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     2701 ccaatagacc gaaatcggca aaatccctta taaatcaaaa gaatagcccg agatagagtt
     2761 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     2821 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac ccaaatcaag
     2881 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     2941 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     3001 agcgggcgct aaggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     3061 cgcgcttaat gcgccgctac agggcgcgta ctatggttgc tttgacgtat gcggtgtgaa
     3121 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg
     3181 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     3241 gggggatgtg ctgcaaggcg attaagttgg gtaacgcca
//
