LOCUS       sequence-342531-        8034 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of dsRed and GFP in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8034)
  AUTHORS   Helaine S, Thompson JA, Watson KG, Liu M, Boyle C, Holden DW
  TITLE     Dynamics of intracellular bacterial replication at the single cell 
            level.
  JOURNAL   Proc Natl Acad Sci U S A. 2010 Feb 23;107(8):3746-51. doi: 
            10.1073/pnas.1000041107. Epub 2010 Feb 2.
  PUBMED    20133586
REFERENCE   2  (bases 1 to 8034)
  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: 
            "10.1073/pnas.1000041107"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2010-02-23- 23"; volume: "107"; issue: "8"; pages: "3746-51"
COMMENT     Sequence Label: pDiGi
FEATURES             Location/Qualifiers
     source          1..8034
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..17
                     /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        join(47..52,53..70,71..77)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    85..101
                     /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)."
     primer_bind     109..125
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     109..125
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             189..902
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(219..247)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     842..862
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             complement(1181..2059)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2086..2370
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     2293..2312
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     primer_bind     complement(2578..2596)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(2628..2648)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2846..2865
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3040..3063
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(3164..3181)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3334..3420
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3512..3539
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3558..3649
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3650..3718,3719..4510)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3868..3887)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4552..5007
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4639..4658)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4849..4870
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5118..5706
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5607..5626
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5860..5877
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6043..6065
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6203..6222
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     protein_bind    6425..6458
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(6610..7692)
                     /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"
     primer_bind     7654..7673
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(7999..8004,8005..8020,8021..8027)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
ORIGIN
        1 ttgtgagcgg ataacaattt cacacaggaa actaggcacc ccaggcttta cactttatgc
       61 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
      121 atgaccatga ttacgaattc ccggattaat taagtgggat cctctagatt taagaaggag
      181 atatacatat gagtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat
      241 tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa
      301 catacggaaa acttaccctt aaatttattt gcactactgg aaaactacct gttccatggc
      361 caacacttgt cactactttc ggttatggtg ttcaatgctt tgcgagatac ccagatcata
      421 tgaaacagca tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaaagaacta
      481 tatttttcaa agatgacggg aactacaaga cacgtgctga agtcaagttt gaaggtgata
      541 cccttgttaa tagaatcgag ttaaaaggta ttgattttaa agaagatgga aacattcttg
      601 gacacaaatt ggaatacaac tataactcac acaatgtata catcatggca gacaaacaaa
      661 agaatggaat caaagttaac ttcaaaatta gacacaacat tgaagatgga agcgttcaac
      721 tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca
      781 accattacct gtccacacaa tctgcccttt cgaaagatcc caacgaaaag agagaccaca
      841 tggtccttct tgagtttgta acagctgctg ggattacaca tggcatggat gaactataca
      901 aataaatgtc cagacctgca ggcatgcaag ctttaatgcg gtagtttatc acagttaaat
      961 tgctaacgca gtcaggcacc gtgtatgaaa tctaacaatg cgctcatcgt catcctcggc
     1021 accgtcaccc tggatgctgt aggcataggc ttggttatgc cggtactgcc gggcctcttg
     1081 cgggatacta tgcataatgt gcctgtcaaa tggacgaagc agggattctg caaaccctat
     1141 gctactccgt caagccgtca attgtctgat tcgttaccaa ttatgacaac ttgacggcta
     1201 catcattcac tttttcttca caaccggcac ggaactcgct cgggctggcc ccggtgcatt
     1261 ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga ccgacggtgg
     1321 cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc
     1381 gccagcttaa gacgctaatc cctaactgct ggcggaaaag atgtgacaga cgcgacggcg
     1441 acaagcaaac atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc
     1501 tgatgtactg acaagcctcg cgtacccgat tatccatcgg tggatggagc gactcgttaa
     1561 tcgcttccat gcgccgcagt aacaattgct caagcagatt tatcgccagc agctccgaat
     1621 agcgcccttc cccttgcccg gcgttaatga tttgcccaaa caggtcgctg aaatgcggct
     1681 ggtgcgcttc atccgggcga aagaaccccg tattggcaaa tattgacggc cagttaagcc
     1741 attcatgcca gtaggcgcgc ggacgaaagt aaacccactg gtgataccat tcgcgagcct
     1801 ccggatgacg accgtagtga tgaatctctc ctggcgggaa cagcaaaata tcacccggtc
     1861 ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg accgcgaatg gtgagattga
     1921 gaatataacc tttcattccc agcggtcggt cgataaaaaa atcgagataa ccgttggcct
     1981 caatcggcgt taaacccgcc accagatggg cattaaacga gtatcccggc agcaggggat
     2041 cattttgcgc ttcagccata cttttcatac tcccgccatt cagagaagaa accaattgtc
     2101 catattgcat cagacattgc cgtcactgcg tcttttactg gctcttctcg ctaaccaaac
     2161 cggtaacccc gcttattaaa agcattctgt aacaaagcgg gaccaaagcc atgacaaaaa
     2221 cgcgtaacaa aagtgtctat aatcacggca gaaaagtcca cattgattat ttgcacggcg
     2281 tcacactttg ctatgccata gcatttttat ccataagatt agcggatcct acctgacgct
     2341 ttttatcgca actctctact gtttctccat acccgttttt ttgggctagc aggaggaatt
     2401 caccatggta cccggggatc ctctagattt aagaaggaga tatacatatg gcctccaccg
     2461 aggacgtcat caaggagttc atgcgcttca aggtgcgcat ggagggctcc gtgaacggcc
     2521 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
     2581 agctgaaggt gaccaagggc ggccccctgc ccttcgcctg ggacatcctg tccccccagt
     2641 tccagtacgg ctccaaggtg tacgtgaagc accccgccga catccccgac tacaagaagc
     2701 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg
     2761 tgaccgtgac ccaggactcc tccctgcagg acggctgctt catctacaag gtgaagttca
     2821 tcggcgtgaa cttcccctcc gacggccccg taatgcagaa gaagactatg ggctgggagc
     2881 cctccaccga gcgcctgtac ccccgcgacg gcgtgctgaa gggcgagatc cacaaggccc
     2941 tgaagctgaa ggacggcggc cactacctgg tggagttcaa gtccatctac atggccaaga
     3001 agcccgtgca gctgcccggc tactactacg tggactccaa gctggacatc acctcccaca
     3061 acgaggacta caccatcgtg gagcagtacg agcgcaccga gggccgctac cacctgttcc
     3121 tgtagaagct tggctgtttt ggcggatgag agaagatttt cagcctgata cagattaaat
     3181 cagaacgcag aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc
     3241 cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt
     3301 ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa
     3361 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
     3421 ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc
     3481 ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt
     3541 gcgtttctac aaactctttt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3601 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     3661 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     3721 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     3781 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     3841 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc
     3901 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     3961 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4021 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4081 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4141 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat
     4201 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4261 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4321 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4381 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4441 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4501 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt tacgcgccct
     4561 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4621 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4681 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4741 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     4801 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4861 tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta taagggattt
     4921 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4981 ttaacaaaat attaacgttt acaatttaaa aggatctagg tgaagatcct ttttgataat
     5041 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5101 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5161 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5221 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     5281 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5341 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5401 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5461 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5521 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5581 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5641 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5701 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5761 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     5821 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     5881 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     5941 atagggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     6001 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     6061 agaggttttc accgtcatca ccgaaacgcg cgaggcagaa ggagatggcg cccaacagtc
     6121 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
     6181 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
     6241 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatctgctca tgtttgacag
     6301 cttatcatcg atgcataaac tgcatccctt aacttgtttt tcgtgtacct attttttgtg
     6361 aatcgactct agaggatccc cgggtaccga gctcgaattc cgatcatatt caataaccct
     6421 taatataact tcgtataatg tatgctatac gaagttatta ggtctgaaga ggagtttacg
     6481 tccagccaag cttgccgtct gaaatggttt cagacggcat gcaggtcgag ctcccgaacg
     6541 ccagcaagac gtagcccagc gcgtcggcca gcttgcaatt cgcgctaact tacattaatt
     6601 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     6661 atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt
     6721 caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag
     6781 caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg
     6841 cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc
     6901 aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc
     6961 aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc
     7021 ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag
     7081 atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa
     7141 cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc
     7201 ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc
     7261 cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt
     7321 aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc
     7381 gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga
     7441 tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc
     7501 aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag
     7561 ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt
     7621 caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt
     7681 tactggtttc acattcaccc cctgaattga ctctcttccg ggcgctatca tgccataccg
     7741 cgaaaggttt tgcaccattc gatggtgtca acgtaaatgc cgcttcgcct tcgcgcgcga
     7801 attgcaagct gatccgggct tatcgactgc acggtgcacc aatgcttctg gcgtcaggca
     7861 gccatcggaa gctgtggtat ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct
     7921 caaggcgcac tcccgttctg gataatgttt tttgcgccga catcataacg gttctggcaa
     7981 atattctgaa atgagctgtt gacaattaat catcggctcg tataatgtgt ggaa
//
