LOCUS       sequence-409116-        5339 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Biosensor for intracellular L-lactate.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5339)
  AUTHORS   Nasu Y, Aggarwal A, Le GNT, Vo CT, Kambe Y, Wang X, Beinlich FRM, 
            Lee AB, Ram TR, Wang F, Gorzo KA, Kamijo Y, Boisvert M, Nishinami S,
            Kawamura G, Ozawa T, Toda H, Gordon GR, Ge S, Hirase H, Nedergaard 
            M, Paquet ME, Drobizhev M, Podgorski K, Campbell RE
  TITLE     Lactate biosensors for spectrally and spatially multiplexed 
            fluorescence imaging.
  JOURNAL   Nat Commun. 2023 Oct 27;14(1):6598. doi: 10.1038/s41467-023-42230-5.
  PUBMED    37891202
REFERENCE   2  (bases 1 to 5339)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2023-10-27"; volume: "14(1):6598. doi"; pages: " 
            10.1038/s41467-023-42230-5"
COMMENT     Sequence Label: pBAD-R-iLACCO1.1
FEATURES             Location/Qualifiers
     source          1..5339
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /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     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             331..348
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             352..384
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(388..396,397..411)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     1449..1468
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1740..1757)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1910..1996
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2088..2115
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2134..2225
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2226..2294,2295..3086)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2444..2463)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3257..3845
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3746..3765
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3999..4016
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4031..4171
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4097..4116)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4257..4279
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4435..5313)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aattaaccat ggggggttct catcatcatc atcatcatgg tatggctagc
      361 atgactggtg gacagcaaat gggtcgggat ctgtacgacg atgacgataa ggatccgagc
      421 tcgagaatgg aacaaatcat agtccagccg ttgaagaccc tcatgggaga cgaccctgat
      481 tattcatttg atattctgga ggcgcggtac gcgatagaag cgtcaactgc ctggtacgct
      541 gcactgcgag ctacacccgg ggacaaagag aaaattcaac tgtgctacga ggctacacgg
      601 tccgaagatc ccgatatagc ttcccaagca gacgtaaggt tccacctggc gattgctgaa
      661 gcaagccata acatagtact gcttcagaca atgcgaggct tttttgatgt tttgcagtct
      721 tcagtcaagc acagtcgaca gcggatgtac ttggattggg tttccgagcg gatgttcccc
      781 gaggacggcg ccctgaagag cgagatcaag aaggggctga ggctgaagga cggcggccac
      841 tacgccgccg aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc
      901 tacgtcgtcg acatcaagtt ggacatcgtg tcccacaacg aggactacac tatcgtggaa
      961 cagtgcgaac gcgccgaggg ccgccactcc accggcggcg tggacgagct gtacaaggga
     1021 ggtacaggcg ggagtctggt gagcaagggc gaggaagatg acatggccat cgtcaaggag
     1081 ttcatgcgct acaaggtgca catggagggc tccgtaaacg gccacgagtt cgagatcgag
     1141 ggcgagggcg agggccgccc ctacgaggcc tttcagaccg ctaagctgaa ggtgaccaag
     1201 ggtggccccc tgcccttcgc ctgggacatc ctatcccctc agttcatgta cggctccaag
     1261 gcctacatta agcacccagc cgacatcccc gactacttca agctgtcctt ccccgagggc
     1321 ttcaggtggg agcgcgtgat gtacttcgag gacggcggca ttattcacgt taaccaggac
     1381 tcctccctgc aggacggcgt attcatctac aaggtgaagc tgcgcggcac caacttcccc
     1441 cccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcgaggcc gccggtctac
     1501 ggtcaactca cagaacagca tcaggctgtt atagacgcca tctttgccgg cgatgcggac
     1561 ggtgctagaa aagctatgat ggctcatctg agcttcgtcc atacgactat gaaacgcttc
     1621 gatggagatc aggcccgaca cgcacgcatt tccagactgc ctggagagca taatgagcac
     1681 tctcgggaaa agaaagcatg aaagcttggc tgttttggcg gatgagagaa gattttcagc
     1741 ctgatacaga ttaaatcaga acgcagaagc ggtctgataa aacagaattt gcctggcggc
     1801 agtagcgcgg tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc
     1861 gatggtagtg tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg
     1921 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     1981 cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg
     2041 gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct
     2101 gacggatggc ctttttgcgt ttctacaaac tcttttgttt atttttctaa atacattcaa
     2161 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2221 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2281 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     2341 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     2401 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     2461 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     2521 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     2581 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     2641 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     2701 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     2761 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     2821 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     2881 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     2941 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3001 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3061 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     3121 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     3181 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     3241 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3301 aacaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3361 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     3421 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3481 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3541 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3601 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3661 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3721 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3781 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     3841 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     3901 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     3961 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     4021 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     4081 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc
     4141 gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc
     4201 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     4261 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agcagatcaa
     4321 ttcgcgcgcg aaggcgaagc ggcatgcata atgtgcctgt caaatggacg aagcagggat
     4381 tctgcaaacc ctatgctact ccgtcaagcc gtcaattgtc tgattcgtta ccaattatga
     4441 caacttgacg gctacatcat tcactttttc ttcacaaccg gcacggaact cgctcgggct
     4501 ggccccggtg cattttttaa atacccgcga gaaatagagt tgatcgtcaa aaccaacatt
     4561 gcgaccgacg gtggcgatag gcatccgggt ggtgctcaaa agcagcttcg cctggctgat
     4621 acgttggtcc tcgcgccagc ttaagacgct aatccctaac tgctggcgga aaagatgtga
     4681 cagacgcgac ggcgacaagc aaacatgctg tgcgacgctg gcgatatcaa aattgctgtc
     4741 tgccaggtga tcgctgatgt actgacaagc ctcgcgtacc cgattatcca tcggtggatg
     4801 gagcgactcg ttaatcgctt ccatgcgccg cagtaacaat tgctcaagca gatttatcgc
     4861 cagcagctcc gaatagcgcc cttccccttg cccggcgtta atgatttgcc caaacaggtc
     4921 gctgaaatgc ggctggtgcg cttcatccgg gcgaaagaac cccgtattgg caaatattga
     4981 cggccagtta agccattcat gccagtaggc gcgcggacga aagtaaaccc actggtgata
     5041 ccattcgcga gcctccggat gacgaccgta gtgatgaatc tctcctggcg ggaacagcaa
     5101 aatatcaccc ggtcggcaaa caaattctcg tccctgattt ttcaccaccc cctgaccgcg
     5161 aatggtgaga ttgagaatat aacctttcat tcccagcggt cggtcgataa aaaaatcgag
     5221 ataaccgttg gcctcaatcg gcgttaaacc cgccaccaga tgggcattaa acgagtatcc
     5281 cggcagcagg ggatcatttt gcgcttcagc catacttttc atactcccgc cattcagag
//
