LOCUS       sequence-353592-        4171 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmids for production of E. coli bacterial 
            ghost (BG) capsules.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4171)
  AUTHORS   Dranchak PK, Oliphant E, Queme B, Lamy L, Wang Y, Huang R, Xia M, 
            Tao D, Inglese J
  TITLE     In vivo quantitative high-throughput screening for drug discovery 
            and comparative toxicology.
  JOURNAL   Dis Model Mech. 2023 Mar 1;16(3):dmm049863. doi: 10.1242/dmm.049863.
            Epub 2023 Mar 20.
  PUBMED    36786055
REFERENCE   2  (bases 1 to 4171)
  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: "Dis Model 
            Mech."; date: "2023-03-1"; volume: "16(3):dmm049863. doi"; pages: " 
            10.1242/dmm.049863. Epub 2023 Mar 20"
COMMENT     Sequence Label: p-lambda-PR cI-Elysis
FEATURES             Location/Qualifiers
     source          1..4171
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(35..748)
                     /codon_start=1
                     /gene="cIts"
                     /product="temperature-sensitive variant of the phage lambda
                     repressor"
                     /label=lambda repressor (ts)
                     /note="thermosensitivity is conferred by the A67T mutation"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLTKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPKLRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     RBS             837..845
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(1296..1314)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1297..1314)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1332..1348)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1332..1348)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1345..1367)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1356..1372
                     /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(1380..1386,1387..1404,1405..1410))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1425..1446
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1563..1580)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1734..2322)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1814..1833)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2493..3284,3285..3353))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3116..3135
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3354..3458)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3536..3991)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3673..3694)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3885..3904
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4117..4139
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4131..4148
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4132..4148
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(4155..4171,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(4155..4171,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggcgaattg ggcccgacca gaacaccttg ccgatcagcc aaacgtctct tcaggccact
       61 gactagcgat aactttcccc acaacggaac aactctcatt gcatgggatc attgggtact
      121 gtgggtttag tggttgtaaa aacacctgac cgctatccct gatcagtttc ttgaaggtaa
      181 actcatcacc cccaagtctg gctatgcaga aatcacctgg ctcaacagcc tgctcagggt
      241 caacgagaat taacattccg tcaggaaagc ttggcttgga gcctgttggt gcggtcatgg
      301 aattaccttc aacctcaagc cagaatgcag aatcactggc ttttttggtt gtgcttaccc
      361 atctctccgc atcacctttg gtaaaggttc taagcttagg tgagaacatc cctgcctgaa
      421 catgagaaaa aacagggtac tcatactcac ttctaagtga cggctgcata ctaaccgctt
      481 catacatctc gtagatttct ctggcgattg aagggctaaa ttcttcaacg ctaactttga
      541 gaatttttgt aagcaatgcg gcgttataag catttaatgc attgatgcca ttaaataaag
      601 caccaacgcc tgactgcccc atccccatct tgtctgcgac agattcctgg gataagccaa
      661 gttcattttt ctttttttca taaattgctt taaggcgacg tgcgtcctca agctgctctt
      721 gtgttaatgg tttctttttt gtgctcatac gttaaatcta tcaccgcaag ggataaatat
      781 ctaacaccgt gcgtgttgac tattttacct ctggcggtga taatggttgc atgtactaag
      841 gaggttgtat ggaacaacgc ataaccctga aagattatgc aatgcgcttt gggcaaacca
      901 agacagctaa agggccgcgg gaattcgatt tatggtacgc tggactttgt gggataccct
      961 cgctttcctg ctcctgttga gtttattgct gccgtcattg cttattatgt tcatcccgtc
     1021 aacattcaaa cggcctgtct catcatggaa ggcgctgaat ttacggaaaa cattattaat
     1081 ggcgtcgagc gtccggttaa agccgctgaa ttgttcgcgt ttaccttgcg tgtacgcgca
     1141 ggaaacactg acgttcttac tgacgcagaa gaaaacgtgc gtcaaaaatt acgtgcggaa
     1201 ggagtgatgt aatgtaatca ctagtgaatt cgcggccgcc tgcaggtcga ccatatggga
     1261 gagctcccaa cgcgttggat gcatagcttg agtattctat agtgtcacct aaatagcttg
     1321 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     1381 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     1441 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     1501 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     1561 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     1621 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     1681 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1741 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     1801 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     1861 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     1921 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     1981 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2041 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2101 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2161 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2221 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2281 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2341 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     2401 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     2461 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     2521 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     2581 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     2641 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     2701 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     2761 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     2821 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     2881 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     2941 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3001 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3061 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     3121 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     3181 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     3241 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     3301 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     3361 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     3421 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     3481 cctgatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt
     3541 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     3601 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     3661 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     3721 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     3781 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     3841 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     3901 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     3961 gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg cgcaactgtt
     4021 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     4081 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     4141 cggccagtga attgtaatac gactcactat a
//
