LOCUS       sequence-262926-        4608 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Positive control for the AVA-seq system. LGF2 (lambda CI associated)
            and Gal11p (RNAp associated) interaction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4608)
  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 4608)
  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-Gal11p
FEATURES             Location/Qualifiers
     source          1..4608
                     /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(3577..3593)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3577..3593)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    3601..3617
                     /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(3625..3631,3632..3649,3650..3655))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             complement(join(4168..4608,1..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 tggtcaaggc tagaaagact ggaacagcta tttctactga tttttcctcg
     2401 agaagacctt gacatgattt tgaaaatgga ttctttacag gatataaaag cattgttaac
     2461 aggattattt ataactgact gattgataat caattactta atcatagctt ggatttttct
     2521 cagggccttt tgcacaatgt atttcctaat aaaactttgc ttgacaggtt ccgtcatcgt
     2581 actacccaat tgtgtttttt caaaattctt gcaggtttcg atggcttctt gtaataagct
     2641 gcgttgctcc tctggagaaa agatttggtt aatttggttg aggacgtttg gtacggtatt
     2701 ttgtgatgat tgctgttgca aagggttgtt gtccgcactt gaattgggag gttgcatttg
     2761 ctgctgttgc caaacgattg gggccgcctc tggtttctct tctttcactt caggctgacg
     2821 tacatcacgt aagtcaacga aagcttccag ttgttcagcc agaatggttg ccgcacgacg
     2881 aatcgcctct tcaggatcga ttgtgccgtt ggtttccatt tcgatgacca gcttgtccag
     2941 gtcggtacgc tgttctacac gcgctgcttc aacattgtag gcaatacgct ccacagggct
     3001 gtagcatgcg tcgaccagca gacggccgat tgggcgctca tcttcttccg aatgaactcg
     3061 ggtagaagcc ggcacataac cacgaccgcg ctgaactttg atacgcatgc taatagacgc
     3121 gttctcatcg gtcaggtggc agatcacgtg ctgcggcttg acgatttcga catcaccgtc
     3181 gtgggtgata tcggctgcag tcacagggcc aatgccagat ttattcaagg taagaataac
     3241 ttcatctttg ccctgaactc tcaccgccag ccctttcagg ttgagcagga tttccaggat
     3301 atcttcctga acgccttctt tggtgctgta ctcatgtagt acaccatcaa tctcaacctc
     3361 ggtcaccgcg caacccggca tcgatgagag cagaatacgg cgcagtgcgt tacccagagt
     3421 atggccaaag ccacgctcta aaggctcaag ggtcaccttg gcgtgcgtcg aactcacttg
     3481 ctcgatatca accaggcgcg gttttagaaa ctctgtcaca gaaccctgca ttgtgtcctc
     3541 tctttggtct agagttccag tgaatccgta atcatggtca tagctgtttc ctgtgtgaaa
     3601 ttgttatccg ctcacaattc cacacattat acgagccgga agcataaagt gtaaagcctc
     3661 tctagctaga ttgagttaat ctccatgtag cgttacaagt ataacacaaa gttttttatg
     3721 ttgtcaatat ttttttgatg ggaaggcact tatttttgac gttcgctcga gttaattaat
     3781 taattaagat cttaggcgcg ccggatctgc atcgcaggat gctgctggct accctgtgga
     3841 acacctacat ctgtattaac gaagcgctaa ccgtttttat caggctctgg gaggcagaat
     3901 aaatgatcat atcgtcaatt attacctcca cggggagagc ctgagcaaac tggcctcagg
     3961 catttgagaa gcacacggtc acactgcttc cggtagtcaa taaaccggta aaccagcaat
     4021 agacataagc ggctatttaa cgaccctgcc ctgaaccgac gaccgggtcg aatttgcttt
     4081 cgaatttctg ccattcatcc gcttattatc acttattcag gcgtagcaac caggcgttta
     4141 agggcaccaa taactgcctt aaaaaaatta cgccccgccc tgccactcat cgcagtactg
     4201 ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg
     4261 aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac
     4321 gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca
     4381 gggattggct gagacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt
     4441 ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg
     4501 gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg
     4561 gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacg
//
