LOCUS       sequence-324111-        3230 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Golden Gate assembly and cell-free (bacterial) expression of 
            fusion-proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3230)
  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 3230)
  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: GGDestX1-Amp
FEATURES             Location/Qualifiers
     source          1..3230
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(328..1119,1120..1188))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     951..970
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1189..1293)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1361..1379
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     RBS             1717..1739
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1789..1810)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2050..2069)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2397..2418
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2628..2645)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(2799..3230,1..157))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2879..2898)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
       61 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
      121 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
      181 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
      241 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
      301 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
      361 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
      421 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
      481 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
      541 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
      601 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
      661 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
      721 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
      781 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
      841 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
      901 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaacacggg ataataccgc
      961 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     1021 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     1081 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     1141 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     1201 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     1261 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     1321 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     1381 ctttcgactt caagaattct ggcgaatcct ctgaccagcc agaaaacgac ctttctgtgg
     1441 tgaaaccgga tgctgcaatt cagagcgcca gcaagtgggg gacagcagaa gtcctgaccg
     1501 ccgcagagtg gatgtttgac atggtgaagt ctatcgcacc atcagccaga aaaccgaatt
     1561 ttgctgggtg ggctaacgat atccgcctga tgcgtgaacg tgacggacgt aaccaccgcg
     1621 acatgtgtgt gctgttccgc tgggcatgct gagctaacac cgtgcgtgtt gacaatttta
     1681 cctctggcgg tgataatggt tgcagctagc aataattttg tttaacttta agaaggagat
     1741 atacatatgg aggggtcttc ggagcttttc actggcgttg ttcccatcct ggtcgagctg
     1801 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     1861 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     1921 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     1981 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     2041 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     2101 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     2161 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     2221 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     2281 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     2341 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     2401 gtcctgctgg agttcgtgac cgccgccggg atcagaagac ctactagcta actcgagcaa
     2461 agcccgccga aaggcgggct tttctgtgtc gaccgatgcc cttgagagcc ttcaacccag
     2521 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgacttct
     2581 ttatcatgca actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac
     2641 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2701 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2761 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2821 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2881 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2941 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3001 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3061 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3121 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3181 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
//
