LOCUS       40924_38458        6169 bp DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses BG4 scFv anti-G-quadruplex antibody in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6169)
  AUTHORS   Biffi G, Tannahill D, McCafferty J, Balasubramanian S
  TITLE     Quantitative visualization of DNA G-quadruplex structures in human 
            cells.
  JOURNAL   Nat Chem. 2013 Mar;5(3):182-6. doi: 10.1038/nchem.1548. Epub 2013 
            Jan 20.
  PUBMED    23422559
REFERENCE   2  (bases 1 to 6169)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6169)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nchem.1548"; journalName: "Nat Chem"; date: "2013-03"; 
            volume: "5"; issue: "3"; pages: "182-6"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6169
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(1..140)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     226..248
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(245..433)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(1208..1229)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(1245..2324)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(2325..2402)
                     /label=lacI promoter
     primer_bind     2608..2627
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI,
                     reverse primer"
     promoter        2711..2729
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2730..2754
                     /label=lac operator
                     /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)."
     RBS             2769..2791
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     sig_peptide     2799..2864
                     /label=pelB signal sequence
                     /note="leader peptide for secretion"
     CDS             3630..3647
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             3654..3719
                     /codon_start=1
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags, followed by an 
                     enterokinase cleavage site"
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3744..3761
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      3828..3875
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3912..4367
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(4462..5274)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      5396..5984
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6138..6155
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact
       61 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
      121 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
      181 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
      241 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
      301 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
      361 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
      421 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac
      481 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac
      541 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca
      601 ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc
      661 agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca
      721 gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt
      781 tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa
      841 ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg
      901 gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag
      961 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca
     1021 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct
     1081 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc
     1141 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg
     1201 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
     1261 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     1321 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg
     1381 cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc
     1441 aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta
     1501 tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg
     1561 cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc
     1621 tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt
     1681 tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga
     1741 cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg
     1801 accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg
     1861 ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca
     1921 gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg
     1981 agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc
     2041 accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc
     2101 gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt
     2161 tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc
     2221 cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag
     2281 acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat
     2341 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg
     2401 tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag
     2461 gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa
     2521 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc
     2581 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
     2641 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat
     2701 cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta
     2761 gaaataattt tgtttaactt taagaaggag atatacatat gaaatacctg ctgccgaccg
     2821 ctgctgctgg tctgctgctc ctcgctgccc agccggccat ggccgaggtg cagctggtgc
     2881 agtctggggc tgaggtgaag aagcctgggg cctcagtgaa ggtctcctgc aaggcttctg
     2941 gttacacctt taccagctat agtatcagct gggtgcgaca ggcccctgga caagggcttg
     3001 agtggatggg atggatcagc gcttacaatg gtaacacaag ctatgcacag aagctccagg
     3061 gcagagtcac gattaccgcg gacaaatcca cgagcacagc ctacatggag ctgagcagcc
     3121 tgagatctga ggacacggcc gtgtattact gtgcgaaggc cgggcaccgt tcagggagat
     3181 ataacaactg gttcgacccc tggggccagg gcaccctggt caccgtctcc tcactcgagg
     3241 gtggaggcgg ttcaggcgga ggtggctctg gcggtggcgc tagccagtct gagctgactc
     3301 agcctccctc ggtgtcagtg gccccaggac agacggccag gatcacctgt ggggaaaaca
     3361 acattggaag taaaaatgtg cactggtacc agcagaagcc aggccaggcc cctgtgctga
     3421 tcatctatag gggtagcaac cggccctctg ggatccctga gcgattctct ggctccaact
     3481 cggggaacac ggccaccctg accatcagca gagcccaagc cggggatgag gctgactatt
     3541 actgccaggt gtttgatagg aggagtgacc atccggtggt gttcggcgga gggaccaagc
     3601 tgaccgtcct aggtgcggcc gcatccgcac atcatcatca ccatcacaag ctggactaca
     3661 aagaccatga cggtgattat aaagatcatg acatcgatta caaggatgac gatgacaagt
     3721 aataaaagct ttaataagtc gagcaccacc accaccacca ctgagatccg gctgctaaca
     3781 aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc
     3841 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat
     3901 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3961 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4021 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4081 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4141 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4201 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4261 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4321 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     4381 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4441 ccgctcatga attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
     4501 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     4561 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     4621 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     4681 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
     4741 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     4801 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
     4861 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     4921 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
     4981 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     5041 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     5101 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
     5161 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     5221 gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcataacacc
     5281 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgaccaaa atcccttaac
     5341 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5401 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5461 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5521 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     5581 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5641 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5701 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5761 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5821 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     5881 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     5941 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6001 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6061 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     6121 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcg
//
