LOCUS       sequence-92596-e        6830 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of dsRed and constitutive expression of GFP in 
            bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6830)
  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 6830)
  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: pDiGc
FEATURES             Location/Qualifiers
     source          1..6830
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             635..1348
                     /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(665..693)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1288..1308
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             complement(1625..2503)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2530..2814
                     /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     2737..2756
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     primer_bind     complement(3022..3040)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(3072..3092)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     3290..3309
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3484..3507
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(3608..3625)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3778..3864
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3956..3983
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4002..4093
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4094..4162,4163..4954)
                     /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(4312..4331)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4996..5451
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5083..5102)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      5562..6150
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6051..6070
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6304..6321
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6336..6476
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6402..6421)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6562..6584
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6723..6742
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ggggatcccg gagttcatgc gtgatgcaat gaaagtgccg ttctacttcg gtgggacctc
       61 actgcttatc gttgttgtcg tgattatgga ctttatggct caagtgcaaa ctctgatgat
      121 gtccagtcag tatgagtctg cattgaagaa ggcgaacctg aaaggctacg gccgttaatt
      181 ggtcgcctga gaagttacgg agagtaaaaa tgaaagttcg tgcttccgtc aagaaattat
      241 gccgtaactg caaaatcgtt aagcgtgatg gtgtcatccg tgtgatttgc agtgccgagc
      301 cgaagcataa acagcgccaa ggctgatttt ttcgcatatt tttcttgcaa agttgggttg
      361 agctggctag attagccagc caatcttttg tatgtctgta cgtttccatt tgagtatcct
      421 gaaaacgggc ttttcagcat ggtacgtaca tattaaatag taggagtgca tagtggcccg
      481 tatagcaggc attaacattc ctgatcagaa acacgccgtg atcgcgttaa cttcgatcta
      541 cggtgtcggc aagacccgtt ctaaagccat cctggctgca gcgggtatcg ctgaaaatgt
      601 taagatcctc tagatttaag aaggagatat acatatgagt aaaggagaag aacttttcac
      661 tggagttgtc ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt
      721 cagtggagag ggtgaaggtg atgcaacata cggaaaactt acccttaaat ttatttgcac
      781 tactggaaaa ctacctgttc catggccaac acttgtcact actttcggtt atggtgttca
      841 atgctttgcg agatacccag atcatatgaa acagcatgac tttttcaaga gtgccatgcc
      901 cgaaggttat gtacaggaaa gaactatatt tttcaaagat gacgggaact acaagacacg
      961 tgctgaagtc aagtttgaag gtgataccct tgttaataga atcgagttaa aaggtattga
     1021 ttttaaagaa gatggaaaca ttcttggaca caaattggaa tacaactata actcacacaa
     1081 tgtatacatc atggcagaca aacaaaagaa tggaatcaaa gttaacttca aaattagaca
     1141 caacattgaa gatggaagcg ttcaactagc agaccattat caacaaaata ctccaattgg
     1201 cgatggccct gtccttttac cagacaacca ttacctgtcc acacaatctg ccctttcgaa
     1261 agatcccaac gaaaagagag accacatggt ccttcttgag tttgtaacag ctgctgggat
     1321 tacacatggc atggatgaac tatacaaata aatgtccaga cctgcaggca tgcaagcttt
     1381 aatgcggtag tttatcacag ttaaattgct aacgcagtca ggcaccgtgt atgaaatcta
     1441 acaatgcgct catcgtcatc ctcggcaccg tcaccctgga tgctgtaggc ataggcttgg
     1501 ttatgccggt actgccgggc ctcttgcggg acgatgcata atgtgcctgt caaatggacg
     1561 aagcagggat tctgcaaacc ctatgctact ccgtcaagcc gtcaattgtc tgattcgtta
     1621 ccaattatga caacttgacg gctacatcat tcactttttc ttcacaaccg gcacggaact
     1681 cgctcgggct ggccccggtg cattttttaa atacccgcga gaaatagagt tgatcgtcaa
     1741 aaccaacatt gcgaccgacg gtggcgatag gcatccgggt ggtgctcaaa agcagcttcg
     1801 cctggctgat acgttggtcc tcgcgccagc ttaagacgct aatccctaac tgctggcgga
     1861 aaagatgtga cagacgcgac ggcgacaagc aaacatgctg tgcgacgctg gcgatatcaa
     1921 aattgctgtc tgccaggtga tcgctgatgt actgacaagc ctcgcgtacc cgattatcca
     1981 tcggtggatg gagcgactcg ttaatcgctt ccatgcgccg cagtaacaat tgctcaagca
     2041 gatttatcgc cagcagctcc gaatagcgcc cttccccttg cccggcgtta atgatttgcc
     2101 caaacaggtc gctgaaatgc ggctggtgcg cttcatccgg gcgaaagaac cccgtattgg
     2161 caaatattga cggccagtta agccattcat gccagtaggc gcgcggacga aagtaaaccc
     2221 actggtgata ccattcgcga gcctccggat gacgaccgta gtgatgaatc tctcctggcg
     2281 ggaacagcaa aatatcaccc ggtcggcaaa caaattctcg tccctgattt ttcaccaccc
     2341 cctgaccgcg aatggtgaga ttgagaatat aacctttcat tcccagcggt cggtcgataa
     2401 aaaaatcgag ataaccgttg gcctcaatcg gcgttaaacc cgccaccaga tgggcattaa
     2461 acgagtatcc cggcagcagg ggatcatttt gcgcttcagc catacttttc atactcccgc
     2521 cattcagaga agaaaccaat tgtccatatt gcatcagaca ttgccgtcac tgcgtctttt
     2581 actggctctt ctcgctaacc aaaccggtaa ccccgcttat taaaagcatt ctgtaacaaa
     2641 gcgggaccaa agccatgaca aaaacgcgta acaaaagtgt ctataatcac ggcagaaaag
     2701 tccacattga ttatttgcac ggcgtcacac tttgctatgc catagcattt ttatccataa
     2761 gattagcgga tcctacctga cgctttttat cgcaactctc tactgtttct ccatacccgt
     2821 ttttttgggc tagcaggagg aattcaccat ggtacccggg gatcctctag atttaagaag
     2881 gagatataca tatggcctcc accgaggacg tcatcaagga gttcatgcgc ttcaaggtgc
     2941 gcatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     3001 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc ctgcccttcg
     3061 cctgggacat cctgtccccc cagttccagt acggctccaa ggtgtacgtg aagcaccccg
     3121 ccgacatccc cgactacaag aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     3181 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggct
     3241 gcttcatcta caaggtgaag ttcatcggcg tgaacttccc ctccgacggc cccgtaatgc
     3301 agaagaagac tatgggctgg gagccctcca ccgagcgcct gtacccccgc gacggcgtgc
     3361 tgaagggcga gatccacaag gccctgaagc tgaaggacgg cggccactac ctggtggagt
     3421 tcaagtccat ctacatggcc aagaagcccg tgcagctgcc cggctactac tacgtggact
     3481 ccaagctgga catcacctcc cacaacgagg actacaccat cgtggagcag tacgagcgca
     3541 ccgagggccg ctaccacctg ttcctgtaga agcttggctg ttttggcgga tgagagaaga
     3601 ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc
     3661 ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc
     3721 gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa
     3781 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     3841 aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg
     3901 cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag
     3961 gccatcctga cggatggcct ttttgcgttt ctacaaactc ttttgtttat ttttctaaat
     4021 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4081 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4141 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4201 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     4261 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     4321 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     4381 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     4441 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4501 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4561 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4621 tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4681 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4741 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4801 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4861 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4921 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4981 actttagatt gatttacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     5041 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     5101 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     5161 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     5221 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     5281 cgttctttaa tagtggactc ttgttccaaa cttgaacaac actcaaccct atctcgggct
     5341 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     5401 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt taaaaggatc
     5461 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5521 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5581 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5641 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5701 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5761 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5821 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5881 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5941 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6001 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6061 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6121 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6181 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6241 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     6301 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     6361 catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc
     6421 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     6481 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     6541 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     6601 cgaaacgcgc gaggcagcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
     6661 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
     6721 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
     6781 acgatgcgtc cggcgtagag gatctgctca tgtttgacag cttatcatcg
//
