LOCUS       sequence-353453-        3797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ORF filtering plasmid (RNAp associated plasmid for pAVA).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3797)
  AUTHORS   Andrews SS, Schaefer-Ramadan S, Al-Thani NM, Ahmed I, Mohamoud YA, 
            Malek JA
  TITLE     High-resolution protein-protein interaction mapping using 
            all-versus-all sequencing (AVA-Seq).
  JOURNAL   J Biol Chem. 2019 Jul 26;294(30):11549-11558. doi: 
            10.1074/jbc.RA119.008792. Epub 2019 Jun 10.
  PUBMED    31182485
REFERENCE   2  (bases 1 to 3797)
  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"; doi: 
            "10.1074/jbc.RA119.008792"; journalName: "J Biol Chem"; date: 
            "2019-07-26- 26"; volume: "294"; issue: "30"; pages: "11549-11558"
COMMENT     Sequence Label: pBORF-AD
FEATURES             Location/Qualifiers
     source          1..3797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(7..12,13..30,31..37)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    45..61
                     /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     50..72
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     69..85
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     69..85
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     104..124
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        106..124
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     161..177
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     161..177
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(211..227)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(212..228)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             346..1134
                     /codon_start=1
                     /gene="bla(M)"
                     /product="beta-lactamase lacking the signal sequence"
                     /label=bla(M)
                     /note="allows cytosolic expression of beta-lactamase"
                     /translation="PETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMST
                     FKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMS
                     DNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATT
                     LRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAA
                     LGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     primer_bind     complement(492..511)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(1154..1173)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1155..1173)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1183..1200)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1183..1199)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1192..1214)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1341..1796
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1428..1447)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1638..1659
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1822..1926
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1927..2721
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             1927..2721
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1981..2000)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2591..2610
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      2892..3480
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3381..3400
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3634..3651
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3768..3789
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
       61 atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta
      121 aagggaacgc gatcgcgagc tccaccgcgg tggcggccgc tctagaacta gtggatcccc
      181 cgggctgcag gaattcgata tcaagcttat cgataccgtc gacctcgagt agtattcaac
      241 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctgcta
      301 caggatctcc aatcgtttgg tgtcgcactc acaggagagt tgcacccaga aacgctggtg
      361 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
      421 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
      481 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
      541 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
      601 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
      661 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
      721 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
      781 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
      841 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
      901 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
      961 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     1021 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     1081 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaaaagctt
     1141 ggtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca
     1201 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     1261 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     1321 cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     1381 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     1441 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     1501 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     1561 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     1621 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     1681 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     1741 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt
     1801 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     1861 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     1921 aagagtatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     1981 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     2041 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     2101 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     2161 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     2221 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     2281 gcaatgcggc ggctgcatac tcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     2341 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     2401 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     2461 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     2521 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     2581 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     2641 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     2701 cttcttgacg agttcttctg actgtcagac caagtttact catatatact ttagattgat
     2761 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     2821 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     2881 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     2941 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3001 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     3061 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     3121 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     3181 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     3241 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     3301 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     3361 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     3421 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     3481 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     3541 ttctttcctg cgttatccac tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     3601 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     3661 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     3721 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     3781 ctcactcatt aggcacc
//
