LOCUS       sequence-410593-        6116 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control biosensor for intracellular L-lactate.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6116)
  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 6116)
  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: "Nat 
            Commun."; date: "2023-10-27"; volume: "14(1):6598. doi"; pages: " 
            10.1038/s41467-023-42230-5"
COMMENT     Sequence Label: pAAV-gfaABC1D-R-diLACCO1
FEATURES             Location/Qualifiers
     source          1..6116
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     primer_bind     895..914
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        895..913
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      938..947
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1978..1997
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2243..2831
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2296..2316)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2714..2725)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2863..3339
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3062..3081)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3379..3519
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3379..3508
                     /label=AAV2 ITR
     rep_origin      3594..4049
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3681..3700)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3891..3912
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4066..4085)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4185..4207
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4245..4263)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4331..4435
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4436..4504,4505..5296)
                     /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"
     rep_origin      5467..6055
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5956..5975
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtggatct aacatatcct ggtgtggagt aggggacgct
      181 gctctgacag aggctcgggg gcctgagctg gctctgtgag ctggggagga ggcagacagc
      241 caggccttgt ctgcaagcag acctggcagc attgggctgg ccgcccccca gggcctcctc
      301 ttcatgccca gtgaatgact caccttggca cagacacaat gttcggggtg ggcacagtgc
      361 ctgcttcccg ccgcacccca gcccccctca aatgccttcc gagaagccca ttgagcaggg
      421 ggcttgcatt gcaccccagc ctgacagcct ggcatcttgg gataaaagca gcacagcccc
      481 ctaggggctg cccttgctgt gtggcgccac cggcggtgga gaacaaggct ctattcagcc
      541 tgtgcccagg aaaggggatc aggggatgcc caggcatgga cagtgggtgg caggggggga
      601 gaggagggct gtctgcttcc cagaagtcca aggacacaaa tgggtgaggg gagagctctc
      661 cccatagctg ggctgcggcc caaccccacc ccctcaggct atgccagggg gtgttgccag
      721 gggcacccgg gcatcgccag tctagcccac tccttcataa agccctcgca tcccaggagc
      781 gagcagagcc agagcaggtt ggagaggaga cgcatcacct ccgctgctcg cggggatcca
      841 ccggtgagct ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac
      901 gactcactat agggagaccc aagctggcta gcgtgccgcc accatggcct cgagaatggt
      961 acaaaacata gtccagccgt tgaagaccct catgggagac gaccctgatt attcatttga
     1021 tattctggag gcgcggtacg cgatagaagc gtctactgcc tggtacgcag cactgcgagc
     1081 tacacccggg gacaaagaga aaattcaaca gtgcttcgag gctacacggt ccgaagatcc
     1141 cgatatagct tcccaagcaa acgtaaggtt ccacctggcg attgctgaag caagccataa
     1201 catagtactg ctacagacga tgcgaggctt ttttgatgtt ttgcagtctt cagtcaagca
     1261 cagtcgacag cggatgtact tggattgggc ttccgagcgg atgttccccg aggacggcgc
     1321 cctgaggagc gtgatcaaga aggggctgag gctgaaggac ggcggccact acgccgccga
     1381 ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acgtcgtcga
     1441 catcaagttg gacatcgtgt cccacaacga ggactacact atcgtggaac agtgcgaacg
     1501 cgccgagggc cgccactcca ccggcggcgt ggacgagctg tacaagggag gtacaggcgg
     1561 gagtctggtg agcaagggcg aggaggatga catggccatc gtcaaggagt tcatgcgcta
     1621 caaggtgcac atggagggct ccgtaaacgg ccacgagttc gagatcgagg gcgagggcga
     1681 gggccgcccc tacgaggcct ttcagaccgc taagctgaag gtgaccaagg gtggccccct
     1741 gcccttcgcc tgggacatcc tatcccctca gttcatgtac ggctccaagg cctacattaa
     1801 gcacccagcc gatatccccg actacttcaa gctgtccttc cccgagggct tcaggtggga
     1861 gcgcgtgatg tacttcgagg acggcggcat tattcacgtt aaccaggact cctccctgca
     1921 ggacggcgta tttatctaca aggtgaagct gcgcggcacc aacttccccc ccgacggccc
     1981 cgtaatgcag aagaagacca tgggctggga ggcgaggccg ccggtctacg gtcaactcac
     2041 agaacagcat caggcagtta tagacgccat ctttgccggc gatgcggacg gtgctagaaa
     2101 agctatgatg gctcatctga gcttcgtcca ttcgactatg caacgcttcg atggagatca
     2161 ggcccgacac gcacgcattt ccagactgcc tggagagcat aatgagcact ctcgggaaaa
     2221 gaaagcatga aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     2281 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     2341 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     2401 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     2461 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     2521 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     2581 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     2641 atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt
     2701 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     2761 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     2821 cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac
     2881 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc
     2941 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg
     3001 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg
     3061 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc
     3121 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc
     3181 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc
     3241 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg
     3301 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg
     3361 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     3421 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     3481 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     3541 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     3601 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3661 tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3721 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3781 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     3841 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3901 gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat
     3961 tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4021 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     4081 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     4141 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     4201 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     4261 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     4321 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4381 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4441 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4501 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4561 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4621 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4681 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4741 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4801 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4861 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4921 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4981 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5041 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5101 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5161 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5221 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5281 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5341 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5401 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5461 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5521 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5581 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5641 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5701 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5761 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5821 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5881 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5941 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6001 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6061 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
