LOCUS       sequence-348363-        4405 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lac inducible, AMP Resistant, cerulean gene on a low copy backbone. 
            Used as a negative control to the pMesh1_CFP plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4405)
  AUTHORS   Buke F, Grilli J, Cosentino Lagomarsino M, Bokinsky G, Tans SJ
  TITLE     ppGpp is a bacterial cell size regulator.
  JOURNAL   Curr Biol. 2022 Feb 28;32(4):870-877.e5. doi: 
            10.1016/j.cub.2021.12.033. Epub 2022 Jan 5.
  PUBMED    34990598
REFERENCE   2  (bases 1 to 4405)
  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: 
            "10.1016/j.cub.2021.12.033"; journalName: "Curr Biol"; date: 
            "2022-02-28- 28"; volume: "32"; issue: "4"; pages: "870-877.e5"
COMMENT     Sequence Label: pCFP
FEATURES             Location/Qualifiers
     source          1..4405
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     939..958
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(978..1055)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1261..1280
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        join(1366..1371,1372..1389,1390..1396)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1404..1420
                     /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)."
     terminator      2202..2273
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2289..2316
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(2344..2361)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2478..3023)
                     /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(2558..2577)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3137..3231)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(3261..4052,4053..4121))
                     /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     3884..3903
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4122..4226)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(join(4300..4405,1..977))
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt
       61 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt
      121 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatgtcc
      181 gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg
      241 ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga
      301 aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga
      361 gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc
      421 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta
      481 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat
      541 aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga
      601 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag
      661 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg
      721 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca
      781 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa
      841 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc
      901 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat
      961 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc
     1021 ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg
     1081 cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc
     1141 aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac
     1201 catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc
     1261 ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac
     1321 gatgcgtccg gcgtagagga tcgagatcgt ttaggcaccc caggctttac actttatgct
     1381 tccggctcgt ataatgtgtg gaattgtgag cggataacaa tttcagaatt caaaagatct
     1441 aggaggaaaa aaatgagcaa gggcgaggag ctgtttactg gtgtggttcc tatcctggtc
     1501 gaactggacg gtgatgtcaa cggtcataag ttcagcgtta gcggcgaggg tgaaggcgac
     1561 gcaacctacg gtaagctgac cctgaagttc atttgtacga cgggtaagct gccggtgcct
     1621 tggcctaccc tggttaccac cctgacctgg ggtgttcaat gcttcgctcg ctatccagac
     1681 cacatgaagc agcacgattt ttttaagagc gccatgccag agggttacgt tcaggagcgc
     1741 accattttct ttaaagatga tggtaattac aagactcgcg cagaggtgaa gtttgaaggc
     1801 gacactctgg tgaatcgtat tgagctgaag ggcatcgatt ttaaggaaga tggtaatatc
     1861 ctgggccata aactggaata taatgcgatt agcgataatg tgtatatcac cgccgataaa
     1921 caaaagaatg gtattaaggc gcattttaaa attcgccata atatcgagga cggctctgtg
     1981 caactggctg accattatca acaaaatacc ccgattggcg acggtcctgt tctgctgccg
     2041 gacaatcact atctgagcac tcaatctaag ctgagcaaag atcctaatga gaagcgcgat
     2101 cacatggtgc tgctggaatt tgtaacggct gctggcatca cgcatggcat ggatgagctg
     2161 tataaataag gatcctaact cgagtaagga tctccaggca tcaaataaaa cgaaaggctc
     2221 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctctactaga
     2281 gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta tacctaggga tatattccgc
     2341 ttcctcgctc actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta
     2401 cgaacggggc ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg
     2461 gccgcggcaa agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga
     2521 cgctcaaatc agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2581 ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg
     2641 ttatggccgc gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc
     2701 caagctggac tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa
     2761 ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg
     2821 taattgattt agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga
     2881 caagttttgg tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc
     2941 agagaacctt cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta
     3001 cgcgcagacc aaaacgatct caagaagatc atcttattaa tcagataaaa tatttctaga
     3061 tttcagtgca atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt
     3121 tgttactagt gcttggattc tcaccaataa aaaacgcccg gcggcaaccg agcgttctga
     3181 acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt ccaagcgagc
     3241 tcgtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3301 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3361 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3421 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3481 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3541 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3601 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3661 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3721 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3781 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3841 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3901 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3961 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4021 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4081 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     4141 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4201 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcggt
     4261 gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg
     4321 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     4381 cgtattgggc gccagggtgg ttttt
//
