LOCUS       sequence-388669-        4984 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express wild type 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
FEATURES             Location/Qualifiers
     source          1..4984
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(6..24)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        92..196
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(197..265,266..1057)
                     /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(415..434)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1228..1816
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1717..1736
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1970..1987
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2104..2125
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2140..2145,2146..2163,2164..2170)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2178..2194
                     /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     2183..2205
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2202..2218
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2202..2218
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     sig_peptide     2270..2335
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     CDS             2711..2728
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2735..2800
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2826..2855
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4110..4127)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4110..4126)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4119..4141)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4339..4794
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4426..4445)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4636..4657
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4811..4830)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4930..4952
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
       61 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
      121 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
      181 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
      241 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
      301 agatcagttg ggtgcccgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
      361 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
      421 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
      481 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
      541 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
      601 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
      661 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
      721 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
      781 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
      841 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
      901 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
      961 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     1021 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     1081 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     1141 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     1201 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     1261 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1321 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     1381 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1441 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1501 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1561 catacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1621 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1681 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1741 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     1801 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     1861 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     1921 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     1981 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     2041 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     2101 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     2161 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     2221 tgattacgcc aagcttgcat gcaaattcta tttcaaggag acagtcataa tgaaatacct
     2281 attgcctacg gcagccgctg gattgttatt actcgcggcc cagccggcca tggcggaagt
     2341 ggagctgcag gcttccgggg gaggatttgt gcagccgggg gggtcattgc gactgagctg
     2401 cgccgcatcc ggcggcacaa gtggtacata caacatgggc tggtttcgtc aggcccctgg
     2461 caaggagaga gagttcgttt ccgccatctc gtaccgtgac aacatgacac cttactacgc
     2521 tgacagcgta aagggaagat ttacaattag ccgggataac tccaaaaaca cggtctatct
     2581 ccagatgaac agcctcaggg ccgaggacac agctacgtat tactgtgcac gttatcaggg
     2641 tcgtctgcgt attcaccagt cgacatactg gggacagggg acgcaggtaa ctgtgagtag
     2701 cgcggccgca catcatcatc accatcacaa gctggactac aaagaccatg acggtgatta
     2761 taaagatcat gacatcgatt acaaggatga cgatgacaag taataaagat ctcaacgggg
     2821 ccgcagaaca gaaacttatc agtgaggagg acttgaatgg ggccgcatag actgttgaaa
     2881 gttgtttagc aaaacctcat acagaaaatt catttactaa cgtctggaaa gacgacaaaa
     2941 ctctagatcg ttacgctaac tatgagggct gtctgtggaa tgctacaggc gttgtggttt
     3001 gtactggtga cgaaactcag tgttacggta catgggttcc tattgggctt gctatccctg
     3061 aaaatgaggg tggtggctct gagggtggcg gttctgaggg tggcggttct gagggtggcg
     3121 gtactaaacc tcctgagtac ggtgatacac ctattccggg ctatacttat atcaaccctc
     3181 tcgacggcac ttatccgcct ggtactgagc aaaaccccgc taatcctaat ccttctcttg
     3241 aggagtctca gcctcttaat actttcatgt ttcagaataa taggttccga aataggcagg
     3301 gtgcattaac tgtttatacg ggcactgtta ctcaaggcac tgaccccgtt aaaacttatt
     3361 accagtacac tcctgtatca tcaaaagcca tgtatgacgc ttactggaac ggtaaattca
     3421 gagactgcgc tttccattct ggctttaatg aggatccatt cgtttgtgaa tatcaaggcc
     3481 aatcgtctga cctgcctcaa cctcctgtca atgctggcgg cggctctggt ggtggttctg
     3541 gtggcggctc tgagggtggc ggctctgagg gtggcggctc tgagggtggc ggttctgagg
     3601 gtggcggctc tgagggtggc ggttccggtg gcggctccgg ttccggtgat tttgattatg
     3661 aaaaaatggc aaacgctaat aagggggcta tgaccgaaaa tgccgatgaa aacgcgctac
     3721 agtctgacgc taaaggcaaa cttgattctg tcgctactga ttacggtgct gctatcgatg
     3781 gtttcattgg tgacgtttcc ggccttgcta atggtaatgg tgctactggt gattttgctg
     3841 gctctaattc ccaaatggct caagtcggtg acggtgataa ttcaccttta atgaataatt
     3901 tccgtcaata tttaccttct ttgcctcagt cggttgaatg tcgcccttat gtctttggcg
     3961 ctggtaaacc atatgaattt tctattgatt gtgacaaaat aaacttattc cgtggtgtct
     4021 ttgcgtttct tttatatgtt gccaccttta tgtatgtatt ttcgacgttt gctaacatac
     4081 tgcgtaataa ggagtcttaa taagaattca ctggccgtcg ttttacaacg tcgtgactgg
     4141 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     4201 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     4261 gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     4321 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4381 ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct
     4441 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4501 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     4561 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4621 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg
     4681 gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc
     4741 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     4801 gcagtctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     4861 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     4921 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     4981 gacg
//
