LOCUS       sequence-262924-        4613 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Negative control for the AVA-seq system. LGF2 (with one nucleotide 
            insertion) (lambda CI associated) and Gal11p (with one nucleotide 
            insertion) (RNAp associated) interaction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4613)
  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 4613)
  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: pAVA-LGF2(fs)-Gal11p(fs)
FEATURES             Location/Qualifiers
     source          1..4613
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     134..153
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(220..322)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(714..731)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(848..1393)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(928..947)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(1556..1561,1562..1579,1580..1586)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1594..1610
                     /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)."
     CDS             1631..2341
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=lambda repressor
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     primer_bind     complement(3582..3598)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3582..3598)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    3606..3622
                     /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)."
     promoter        complement(join(3630..3636,3637..3654,3655..3660))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             complement(join(4173..4613,1..219))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 gaactccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt
       61 gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt
      121 ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga
      181 tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga
      241 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
      301 ggaacctctt acgtgccgat caacgtctca ttttcgccaa aagttggccc agggcttccc
      361 ggtatcaaca gggacaccag gatttattta ttctgcgaag tgatcttccg tcacaggtat
      421 ttattcggcg caaagtgcgt cgggtgatgc tgccaactta ctgatttagt gtatgatggt
      481 gtttttgagg tgctccagtg gcttctgttt ctatcagctg tccctcctgt tcagctactg
      541 acggggtggt gcgtaacggc aaaagcaccg ccggacatca gcgctagcgg agtgtatact
      601 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa
      661 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc
      721 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc
      781 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa
      841 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
      901 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc
      961 tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc
     1021 gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac
     1081 tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt
     1141 gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt
     1201 agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg
     1261 tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt
     1321 cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc
     1381 aaaacgatct caagaagatc atcttattaa tcagataaaa tatttctaga tttcagtgca
     1441 atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt tgtaattctc
     1501 atgtttgaca gcttatcatc gataagctaa ttctcactca ttaggcaccc caggctttac
     1561 actttatgct tccggctcgt ataatgtgtg gaattgtgag cggataacaa tttcacacag
     1621 gaaacagcgt atgagcacaa aaaagaaacc attaacacaa gagcagcttg aggacgcacg
     1681 tcgccttaaa gcaatttatg aaaaaaagaa aaatgaactt ggcttatccc aggaatctgt
     1741 cgcagacaag atggggatgg ggcagtcagg cgttggtgct ttatttaatg gcatcaatgc
     1801 attaaatgct tataacgccg cattgcttgc aaaaattctc aaagttagcg ttgaagaatt
     1861 tagcccttca atcgccagag aaatctacga gatgtatgaa gcggttagta tgcagccgtc
     1921 acttagaagt gagtatgagt accctgtttt ttctcatgtt caggcaggga tgttctcacc
     1981 tgagcttaga acctttacca aaggtgatgc ggagagatgg gtaagcacaa ccaaaaaagc
     2041 cagtgattct gcattctggc ttgaggttga aggtaattcc atgaccgcac caacaggctc
     2101 caagccaagc tttcctgacg gaatgttaat tctcgttgac cctgagcagg ctgttgagcc
     2161 aggtgatttc tgcatagcca gacttggggg tgatgagttt accttcaaga aactgatcag
     2221 ggatagcggt caggtgtttt tacaaccact aaacccacag tacccaatga tcccatgcaa
     2281 tgagagttgt tccgttgtgg ggaaagttat cgctagtcag tggcctgaag agacgtttgg
     2341 cccacaaggg tggatcaagg ctagaaagac tggaacagct atttctactg atttttcctc
     2401 gagaagacct tgacatgatt ttgaaaatgg attctttaca ggatataaaa gcattgttaa
     2461 caggattatt tataactgac tgattgataa tcaattactt aatcatagct tggatttttc
     2521 tcagggcctt ttgcacaatg tatttcctaa taaaactttg cttgacaggt tccgtcatcg
     2581 tactacccaa ttgtgttttt tcaaaattct tgcaggtttc gatggcttct tgtaataagc
     2641 tgcgttgctc ctctggagaa aagatttggt taatttggtt gaggacgttt ggtacggtat
     2701 tttgtgatga ttgctgttgc aaagggttgt tgtccgcact tgaattggga ggttgcattt
     2761 gctgctgttg aggtccaaac gattggggcc gcctctggtt tctcttcttt cacttcaggc
     2821 tgacgtacat cacgtaagtc aacgaaagct tccagttgtt cagccagaat ggttgccgca
     2881 cgacgaatcg cctcttcagg atcgattgtg ccgttggttt ccatttcgat gaccagcttg
     2941 tccaggtcgg tacgctgttc tacacgcgct gcttcaacat tgtaggcaat acgctccaca
     3001 gggctgtagc atgcgtcgac cagcagacgg ccgattgggc gctcatcttc ttccgaatga
     3061 actcgggtag aagccggcac ataaccacga ccgcgctgaa ctttgatacg catgctaata
     3121 gacgcgttct catcggtcag gtggcagatc acgtgctgcg gcttgacgat ttcgacatca
     3181 ccgtcgtggg tgatatcggc tgcagtcaca gggccaatgc cagatttatt caaggtaaga
     3241 ataacttcat ctttgccctg aactctcacc gccagccctt tcaggttgag caggatttcc
     3301 aggatatctt cctgaacgcc ttctttggtg ctgtactcat gtagtacacc atcaatctca
     3361 acctcggtca ccgcgcaacc cggcatcgat gagagcagaa tacggcgcag tgcgttaccc
     3421 agagtatggc caaagccacg ctctaaaggc tcaagggtca ccttggcgtg cgtcgaactc
     3481 acttgctcga tatcaaccag gcgcggtttt agaaactctg tcacagaacc ctgcattgtg
     3541 tcctctcttt ggtctagagt tccagtgaat ccgtaatcat ggtcatagct gtttcctgtg
     3601 tgaaattgtt atccgctcac aattccacac attatacgag ccggaagcat aaagtgtaaa
     3661 gcctctctag ctagattgag ttaatctcca tgtagcgtta caagtataac acaaagtttt
     3721 ttatgttgtc aatatttttt tgatgggaag gcacttattt ttgacgttcg ctcgagttaa
     3781 ttaattaatt aagatcttag gcgcgccgga tctgcatcgc aggatgctgc tggctaccct
     3841 gtggaacacc tacatctgta ttaacgaagc gctaaccgtt tttatcaggc tctgggaggc
     3901 agaataaatg atcatatcgt caattattac ctccacgggg agagcctgag caaactggcc
     3961 tcaggcattt gagaagcaca cggtcacact gcttccggta gtcaataaac cggtaaacca
     4021 gcaatagaca taagcggcta tttaacgacc ctgccctgaa ccgacgaccg ggtcgaattt
     4081 gctttcgaat ttctgccatt catccgctta ttatcactta ttcaggcgta gcaaccaggc
     4141 gtttaagggc accaataact gccttaaaaa aattacgccc cgccctgcca ctcatcgcag
     4201 tactgttgta attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga
     4261 acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatggtg
     4321 aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc
     4381 acccagggat tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc
     4441 aggttttcac cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg
     4501 tcgtggtatt cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa
     4561 caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgccat acg
//
