LOCUS pHEN2-SG4 4984 bp DNA circular SYN 25-AUG-2024
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 Aug 25, 2024 from SnapGene Server 7.0.3
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Am Chem
Soc. 2022 Dec 9. doi: 10.1021/jacs.2c10656."
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 197..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 2140..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
//
