LOCUS       sequence-484675-        3659 bp    DNA     circular SYN 26-AUG-2025
DEFINITION  PSP-LacO Cassette; AP Prop Circuit for AraC phage-assisted 
            evolution.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3659)
  TITLE     Phage-Assisted Evolution of Allosteric Protein Switches
REFERENCE   2  (bases 1 to 3659)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 26, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: PACE_AP_GIII_Titrate
FEATURES             Location/Qualifiers
     source          1..3659
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(17..676)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAKCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     591..610
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(824..841)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      994..1080
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1172..1199
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     protein_bind    1363..1382
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     CDS             1452..2726
                     /codon_start=1
                     /gene="III"
                     /product="pIII"
                     /label=M13 gene III
                     /note="minor coat protein"
                     /translation="MKKLLFAIPLVVPFYSHSAETVESCLAKPHTENSFTNVWKDDKTL
                     DRYANYEGCLWNATGVVVCTGDETQCYGTWVPIGLAIPENEGGGSEGGGSEGGGSEGGG
                     TKPPEYGDTPIPGYTYINPLDGTYPPGTEQNPANPNPSLEESQPLNTFMFQNNRFRNRQ
                     GALTVYTGTVTQGTDPVKTYYQYTPVSSKAMYDAYWNGKFRDCAFHSGFNEDPFVCEYQ
                     GQSSDLPQPPVNAGGGSGGGSGGGSEGGGSEGGGSEGGGSEGGGSGGGSGSGDFDYEKM
                     ANANKGAMTENADENALQSDAKGKLDSVATDYGAAIDGFIGDVSGLANGNGATGDFAGS
                     NSQMAQVGDGDNSPLMNNFRQYLPSLPQSVECRPFVFGAGKPYEFSIDCDKINLFRGVF
                     AFLLYVATFMYVFSTFANILRNKES"
     rep_origin      2883..3621
                     /direction=RIGHT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
ORIGIN
        1 acggcctatt tggcttttac gccccgccct gccactcatc gcagtactgt tgtaattcat
       61 taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg
      121 gcatcagcac cttgtcgcct tgcgtataat atttgcccat agtgaaaacg ggggcgaaga
      181 agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg
      241 agacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac
      301 acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc
      361 agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat
      421 cccatatcac cagctcaccg tctttcattg ccatacggaa ctccggatga gcattcatca
      481 ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct
      541 ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattta gcaactgact
      601 gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag
      661 tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata
      721 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccacagtc
      781 agtagggccc tggctgtttt ggcggatgag agaagatttt cagcctgata cagattaaat
      841 cagaacgcag aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc
      901 cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt
      961 ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa
     1021 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
     1081 ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc
     1141 ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt
     1201 gcgtttctac aaactcgctg aggacattgg ctccgtgatg aaattcgcca cttgttagtg
     1261 taattcgcta actcatcctg gcatgttgct gttgattctt caatcagatc tttataaatc
     1321 aaaaagataa aaaattggca cgcaaattgt attaacagtt caaattgtga gcgctcacaa
     1381 ttgcaggaca atcctgaacg cagaaatcaa gaggacaaca ttacctgcag gtgcagtaaa
     1441 ggaaaaaaaa aatgaaaaaa ttattattcg caattccttt agttgttcct ttctattctc
     1501 actccgctga aactgttgaa agttgtttag caaaacccca tacagaaaat tcatttacta
     1561 acgtctggaa agacgacaaa actttagatc gttacgctaa ctatgagggc tgtctgtgga
     1621 atgctacagg cgttgtagtt tgtactggtg acgaaactca gtgttacggt acatgggttc
     1681 ctattgggct tgctatccct gaaaatgagg gtggtggctc tgagggtggc ggttctgagg
     1741 gtggcggttc tgagggtggc ggtactaaac ctcctgagta cggtgataca cctattccgg
     1801 gctatactta tatcaaccct ctcgacggca cttatccgcc tggtactgag caaaaccccg
     1861 ctaatcctaa tccttctctt gaggagtctc agcctcttaa tactttcatg tttcagaata
     1921 ataggttccg aaataggcag ggggcattaa ctgtttatac gggcactgtt actcaaggca
     1981 ctgaccccgt taaaacttat taccagtaca ctcctgtatc atcaaaagcc atgtatgacg
     2041 cttactggaa cggtaaattc agagactgcg ctttccattc tggctttaat gaggatccat
     2101 tcgtttgtga atatcaaggc caatcgtctg acctgcctca acctcctgtc aatgctggcg
     2161 gcggctctgg tggtggttct ggtggcggct ctgagggtgg tggctctgag ggtggcggtt
     2221 ctgagggtgg cggctctgag ggaggcggtt ccggtggtgg ctctggttcc ggtgattttg
     2281 attatgaaaa gatggcaaac gctaataagg gggctatgac cgaaaatgcc gatgaaaacg
     2341 cgctacagtc tgacgctaaa ggcaaacttg attctgtcgc tactgattac ggtgctgcta
     2401 tcgatggttt cattggtgac gtttccggcc ttgctaatgg taatggtgct actggtgatt
     2461 ttgctggctc taattcccaa atggctcaag tcggtgacgg tgataattca cctttaatga
     2521 ataatttccg tcaatattta ccttccctcc ctcaatcggt tgaatgtcgc ccttttgtct
     2581 ttggcgctgg taaaccttac gagttcagta tcgactgcga taagatcaac ctgttccgcg
     2641 gtgtctttgc gtttctttta tatgttgcca cctttatgta tgtattttct acgtttgcta
     2701 acatactgcg taataaggag tcttaaaggg tccctaagtc tcctcagcaa aacgaaaggc
     2761 ccagtctttc gactgagcct ttcgttttat ttgaccggat gtcctcttcc tcctgggctg
     2821 acttcaggtg ctacatttga agagataaat tgcactgaaa tctagagcgg ttcagtagaa
     2881 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatcttttg ccctgtaaac
     2941 gaaaaaacca cctggggagg tggtttgatc gaaggttaag tcagttgggg aactgcttaa
     3001 ccgtggtaac tggctttcgc agagcacagc aaccaaatct gtccttccag tgtagccgga
     3061 ctttggcgca cacttcaaga gcaaccgcgt gtttagctaa acaaatcctc tgcgaactcc
     3121 cagttaccaa tggctgctgc cagtggcgtt ttaccgtgct tttccgggtt ggactcaagt
     3181 gaacagttac cggataaggc gcagcagtcg ggctgaacgg ggagttcttg cttacagccc
     3241 agcttggagc gaacgaccta caccgagccg agataccagt gtgtgagcta tgagaaagcg
     3301 ccacacttcc cgtaagggag aaaggcggaa caggtatccg gtaaacggca gggtcggaac
     3361 aggagagcgc aagagggagc gacccgccgg aaacggtggg gatctttaag tcctgtcggg
     3421 tttcgcccgt actgtcagat tcatggttga gcctcacggc tcccacagat gcaccggaaa
     3481 agcgtctgtt tatgtgaact ctggcaggag ggcggagcct atggaaaaac gccaccggcg
     3541 cggccctgct gttttgcctc acatgttagt cccctgctta tccacggaat ctgtgggtaa
     3601 ctttgtatgt gtccgcagcg cccgccgcag tctcacgccc ggagcgtagc gaccgagtg
//
