LOCUS       sequence-388449-        4984 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mutant (R105A) SG4 in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4984)
  AUTHORS   Galli S, Melidis L, Flynn SM, Varshney D, Simeone A, Spiegel J, 
            Madden SK, Tannahill D, Balasubramanian S
  TITLE     DNA G-Quadruplex Recognition In Vitro and in Live Cells by a 
            Structure-Specific Nanobody.
  JOURNAL   J Am Chem Soc. 2022 Dec 9. doi: 10.1021/jacs.2c10656.
  PUBMED    36488193
REFERENCE   2  (bases 1 to 4984)
  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: "J Am Chem 
            Soc. 2022 Dec 9. doi: 10.1021/jacs.2c10656."
COMMENT     Sequence Label: pHEN2-SG4 R105A
FEATURES             Location/Qualifiers
     source          1..4984
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        82..186
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(187..255,256..1047)
                     /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(405..424)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1218..1806
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1707..1726
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1960..1977
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2094..2115
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2130..2135,2136..2153,2154..2160)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2168..2184
                     /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)."
     primer_bind     2173..2195
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2192..2208
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2192..2208
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     sig_peptide     2260..2325
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     CDS             2701..2718
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2725..2790
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2816..2845
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4100..4117)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4100..4116)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4109..4131)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4329..4784
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4416..4435)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4626..4647
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4801..4820)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4920..4942
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(join(4980..4984,1..14))
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
       61 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
      121 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
      181 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
      241 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
      301 ggtgcccgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
      361 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
      421 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
      481 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
      541 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
      601 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
      661 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
      721 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
      781 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
      841 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
      901 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
      961 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     1021 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     1081 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     1141 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1201 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1261 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     1321 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     1381 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     1441 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     1501 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg catacagccc
     1561 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     1621 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     1681 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     1741 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     1801 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     1861 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     1921 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     1981 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     2041 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     2101 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     2161 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     2221 aagcttgcat gcaaattcta tttcaaggag acagtcataa tgaaatacct attgcctacg
     2281 gcagccgctg gattgttatt actcgcggcc cagccggcca tggcggaagt ggagctgcag
     2341 gcttccgggg gaggatttgt gcagccgggg gggtcattgc gactgagctg cgccgcatcc
     2401 ggcggcacaa gtggtacata caacatgggc tggtttcgtc aggcccctgg caaggagaga
     2461 gagttcgttt ccgccatctc gtaccgtgac aacatgacac cttactacgc tgacagcgta
     2521 aagggaagat ttacaattag ccgggataac tccaaaaaca cggtctatct ccagatgaac
     2581 agcctcaggg ccgaggacac agctacgtat tactgtgcac gttatcaggg tgctctgcgt
     2641 attcaccagt cgacatactg gggacagggg acgcaggtaa ctgtgagtag cgcggccgca
     2701 catcatcatc accatcacaa gctggactac aaagaccatg acggtgatta taaagatcat
     2761 gacatcgatt acaaggatga cgatgacaag taataaagat ctcaacgggg ccgcagaaca
     2821 gaaacttatc agtgaggagg acttgaatgg ggccgcatag actgttgaaa gttgtttagc
     2881 aaaacctcat acagaaaatt catttactaa cgtctggaaa gacgacaaaa ctctagatcg
     2941 ttacgctaac tatgagggct gtctgtggaa tgctacaggc gttgtggttt gtactggtga
     3001 cgaaactcag tgttacggta catgggttcc tattgggctt gctatccctg aaaatgaggg
     3061 tggtggctct gagggtggcg gttctgaggg tggcggttct gagggtggcg gtactaaacc
     3121 tcctgagtac ggtgatacac ctattccggg ctatacttat atcaaccctc tcgacggcac
     3181 ttatccgcct ggtactgagc aaaaccccgc taatcctaat ccttctcttg aggagtctca
     3241 gcctcttaat actttcatgt ttcagaataa taggttccga aataggcagg gtgcattaac
     3301 tgtttatacg ggcactgtta ctcaaggcac tgaccccgtt aaaacttatt accagtacac
     3361 tcctgtatca tcaaaagcca tgtatgacgc ttactggaac ggtaaattca gagactgcgc
     3421 tttccattct ggctttaatg aggatccatt cgtttgtgaa tatcaaggcc aatcgtctga
     3481 cctgcctcaa cctcctgtca atgctggcgg cggctctggt ggtggttctg gtggcggctc
     3541 tgagggtggc ggctctgagg gtggcggctc tgagggtggc ggttctgagg gtggcggctc
     3601 tgagggtggc ggttccggtg gcggctccgg ttccggtgat tttgattatg aaaaaatggc
     3661 aaacgctaat aagggggcta tgaccgaaaa tgccgatgaa aacgcgctac agtctgacgc
     3721 taaaggcaaa cttgattctg tcgctactga ttacggtgct gctatcgatg gtttcattgg
     3781 tgacgtttcc ggccttgcta atggtaatgg tgctactggt gattttgctg gctctaattc
     3841 ccaaatggct caagtcggtg acggtgataa ttcaccttta atgaataatt tccgtcaata
     3901 tttaccttct ttgcctcagt cggttgaatg tcgcccttat gtctttggcg ctggtaaacc
     3961 atatgaattt tctattgatt gtgacaaaat aaacttattc cgtggtgtct ttgcgtttct
     4021 tttatatgtt gccaccttta tgtatgtatt ttcgacgttt gctaacatac tgcgtaataa
     4081 ggagtcttaa taagaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     4141 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     4201 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc
     4261 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     4321 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4381 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
     4441 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4501 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     4561 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4621 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
     4681 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
     4741 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcagtctcag
     4801 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     4861 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     4921 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     4981 cctc
//
