LOCUS       sequence-319732-        3284 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Golden Gate assembly of a pTXB1 compatible fusion-protein ORF 
            insert.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3284)
  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 3284)
  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: GGDest3-Amp
FEATURES             Location/Qualifiers
     source          1..3284
                     /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(231..247)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(231..247)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(244..266)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    255..271
                     /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(279..285,286..303,304..309))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    324..345
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(462..479)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(633..1221)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(713..732)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1392..2183,2184..2252))
                     /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     2015..2034
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2253..2357)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2425..2443
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2481..2503)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2603..2622
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2639..3094)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2776..2797)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2988..3007
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattggaggc tacagtcagt
       61 ggagaggact ttcactgact gactgactgc gtctcaacct gaattccata tgaaggaggg
      121 gtcttcgaga agacctacta ggtgccgaag agcctgcaga gcaagagacg actgactgac
      181 tgactggaaa gaggaagggc tggaagagga aggagcttgg cgtaatcatg gtcatagctg
      241 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
      301 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
      361 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
      421 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
      481 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
      541 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
      601 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
      661 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
      721 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
      781 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
      841 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
      901 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
      961 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1021 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     1081 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     1141 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1201 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     1261 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     1321 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     1381 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     1441 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     1501 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     1561 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     1621 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     1681 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     1741 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     1801 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     1861 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     1921 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     1981 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     2041 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     2101 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     2161 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     2221 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     2281 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     2341 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     2401 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     2461 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     2521 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     2581 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccataaa
     2641 attgtaaacg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     2701 tttaaccaat agaccgaaat cggcaaaatc ccttataaat caaaagaata gcccgagata
     2761 gagttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     2821 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcacccaaa
     2881 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     2941 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     3001 aaaggagcgg gcgctaaggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     3061 cccgccgcgc ttaatgcgcc gctacagggc gcgtactatg gttgctttga cgtatgcggt
     3121 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca
     3181 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     3241 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gcca
//
