LOCUS       sequence-319736-        3302 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Golden Gate assembly of a pcDNA3.1+ or BioBrick RFC23 compatible 
            fusion-protein ORF insert.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3302)
  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 3302)
  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: GGDest1-Amp
FEATURES             Location/Qualifiers
     source          1..3302
                     /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"
     regulatory      128..137
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    157..177
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(249..265)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(249..265)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(262..284)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    273..289
                     /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(297..303,304..321,322..327))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    342..363
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(480..497)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(651..1239)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(731..750)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1410..2201,2202..2270))
                     /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     2033..2052
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2271..2375)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2443..2461
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2499..2521)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2621..2640
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2657..3112)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2794..2815)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3006..3025
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattggaggc tacagtcagt
       61 ggagaggact ttcactgact gactgactgc gtctcaacct gaattcgcgg ccgcatctag
      121 acccgccgcc accatggagg ggtcttcgag aagacctact agtagcggcc gctgcagagc
      181 aagagacgac tgactgactg actggaaaga ggaagggctg gaagaggaag gagcttggcg
      241 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
      301 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
      361 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
      421 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
      481 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
      541 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
      601 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
      661 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
      721 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
      781 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
      841 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
      901 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
      961 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     1021 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     1081 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     1141 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     1201 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     1261 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     1321 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     1381 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     1441 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     1501 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     1561 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     1621 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     1681 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     1741 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     1801 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     1861 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     1921 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     1981 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     2041 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     2101 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     2161 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     2221 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     2281 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     2341 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     2401 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     2461 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     2521 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     2581 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     2641 ctgagagtgc accataaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt
     2701 tgttaaatca gctcattttt taaccaatag accgaaatcg gcaaaatccc ttataaatca
     2761 aaagaatagc ccgagataga gttgagtgtt gttccagttt ggaacaagag tccactatta
     2821 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     2881 cgtgaaccat cacccaaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     2941 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     3001 aaggaaggga agaaagcgaa aggagcgggc gctaaggcgc tggcaagtgt agcggtcacg
     3061 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtactatggt
     3121 tgctttgacg tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     3181 ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     3241 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     3301 ca
//
