LOCUS       sequence-218361-        2213 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Promoter repressed by LacI (not natively produced in B. subtilis 
            168).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2213)
  AUTHORS   Wicke N, Radford D, Verrone V, Wipat A, French CE
  TITLE     BacilloFlex: A modular DNA assembly toolkit for Bacillus subtilis 
            synthetic biology
  JOURNAL   bioRxiv 185108
REFERENCE   2  (bases 1 to 2213)
  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"; journalName: "bioRxiv 
            185108"
COMMENT     Sequence Label: pBP-Pspac
FEATURES             Location/Qualifiers
     source          1..2213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      19..76
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(100..117)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(271..859)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(351..370)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1042..1136)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1157..1816)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1731..1750
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     1978..1996
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2000..2043)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2046..2065
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     protein_bind    2173..2192
                     /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."
ORIGIN
        1 ccgtctcatt agctgcagtc cggcaaaaaa gggcaaggtg tcaccaccct gccctttttc
       61 tttaaaaccg aaaagattac ttcgcgttat gcaggcttcc tcgctcactg actcgctgcg
      121 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
      181 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
      241 gaaccgtaaa aaggccgcgt tgctggcgtt tttccacagg ctccgccccc ctgacgagca
      301 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
      361 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
      421 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
      481 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
      541 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
      601 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
      661 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
      721 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
      781 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
      841 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
      901 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
      961 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     1021 gtctgacagc tcgaggcttg gattctcacc aataaaaaac gcccggcggc aaccgagcgt
     1081 tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca ggagtccaag
     1141 cgagctcgat atcaaattac gccccgccct gccactcatc gcagtactgt tgtaattcat
     1201 taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg
     1261 gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga
     1321 agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg
     1381 acacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac
     1441 acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc
     1501 agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat
     1561 cccatatcac cagctcaccg tctttcattg ccatacgaaa ttccggatga gcattcatca
     1621 ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct
     1681 ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact
     1741 gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag
     1801 tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata
     1861 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgcccgatca
     1921 actcgagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     1981 aggcgtatca cgaggcagaa tttcagataa aaaaaatcct tagctttcgc taaggatgat
     2041 ttctggaatt cgcggccgct tctagaatct agagacgcat aggtctcact atagaacaac
     2101 ctctgctaaa attcctgaaa aattttgcaa aaagttgttg actttatcta caaggtgtgg
     2161 cataatgtgt ggaattgtga gcgctcacaa ttaagcttgt acagagaccc atg
//
