LOCUS       sequence-371362-        6806 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To explore endoplasmic reticulum lactate dynamics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6806)
  AUTHORS   Aburto C, Galaz A, Bernier A, Sandoval PY, Holtheuer-Gallardo S, 
            Ruminot I, Soto-Ojeda I, Hertenstein H, Schweizer JA, Schirmeier S, 
            Pastor TP, Mardones GA, Barros LF, San Martin A
  TITLE     Single-Fluorophore Indicator to Explore Cellular and Sub-cellular 
            Lactate Dynamics.
  JOURNAL   ACS Sens. 2022 Oct 28. doi: 10.1021/acssensors.2c00731.
  PUBMED    36306435
REFERENCE   2  (bases 1 to 6806)
  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: "ACS Sens. 
            2022 Oct 28. doi: 10.1021/acssensors.2c00731."
COMMENT     Sequence Label: ER-CanlonicSF
FEATURES             Location/Qualifiers
     source          1..6806
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        12..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     544..564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      650..659
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     join(656..700,783..794)
                     /codon_start=1
                     /product="modified signal sequence from the mouse 
                     immunoglobulin heavy chain variable region"
                     /label=IgH signal sequence
                     /translation="MGWSCIILFLVATATGAHS"
     CDS             join(656..701,784..794)
                     /codon_start=1
                     /product="signal sequence from the mouse immunoglobulin 
                     heavy chain variableregion"
                     /label=IgH signal sequence
                     /translation="MGWSCIILFLVATATGAHS"
     CDS             1371..1622
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGPVLLPDNH
                     YLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     1559..1580
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1686..1707)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1947..1966)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2580..2609
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2663..2680)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2669..2893
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2939..3367
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3026..3045)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3236..3257
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3376..3396)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3381..3710
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3561..3696
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3623..3642
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3736..4530
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MVEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3790..3809)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4400..4419
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4704..4825
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4741..4760)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4795..4814
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4858..4875)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5029..5617)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5109..5128)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5788..6579,6580..6648))
                     /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     6411..6430
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6649..6753)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtaccgaatt cacattgatt attgagtagt tattaatagt aatcaattac ggggtcatta
       61 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      121 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      181 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      241 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      301 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      361 atctacggtt agtcatcgct attaccatag tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctttctggc
      601 taactagaga acccactgct tactggcacg tggaaattaa tacgacgtgg ccaccatggg
      661 atggagctgt atcatcctct tcttggtagc aacagctaca ggtaaggggt taacagtagc
      721 aggcttgagg tctggacata tatatgggtg acaatgacat ccactttgcc tttctctcca
      781 caggcgcgca ctcccaggtc caactgcagg gatccatgtt ttctcctctg gcggtggccc
      841 aagcgaggcg ctaccgttgg cgcatccaga ccgcttggga tgcgggcacg gtaggctatt
      901 ccctgttcca gaagttcacc gagcgtgtta aggagctcac cgacggccag ttggaggttc
      961 agcccttccc cgcaggggcc gtggtgggca cctttgacat gtttgacgcg gtgaagacgg
     1021 gcgttttgga cgggatgaac cccttcaccc tcgcatgggc ggggcggatg ccggtgacgg
     1081 cctttctttc ctcctatgcc ctgggtttgg accggcccga ccagtgggag acctggttct
     1141 acagcctggg cggcctggac atcgcccgga gggccttcgc cgagcagggg cttttctacg
     1201 tgggccccgt gcaacacgat ttgaacatca tccactccaa aaagccgatc cgtcgctttg
     1261 aggatttcaa aggggtaaag ctgcgggtgc cgggcggaat gattgctgaa gtctttgccg
     1321 ccgcgggggc ctccacggtc cttctactag aaaacgtcta tatcaaggcc gacaagcaga
     1381 agaacggcat caaggcgaac ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc
     1441 tcgcctacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1501 accactacct gagcgtccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca
     1561 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     1621 agggtggtac cggtggatct atggtgagca agggcgagga gctgttcacc ggggtggtgc
     1681 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     1741 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     1801 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     1861 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctaca
     1921 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     1981 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     2041 acggcaacat cctggggcac aagctggagt acaacaccag acccggaggc gaggtctatc
     2101 ctgccttgga gcgtggggtc attgatgcgg ccgactttgt ggggcctgcg gtcaactaca
     2161 atctgggctt ccaccaggtg gccaagtaca tcattatggg gcccccggag accccggcca
     2221 tccatcagcc ggtggacctt atggacttca ccatcaacct caatcggtgg cggtcgctac
     2281 ccaagccttt gcaagagcgt ttcatcgccg ctgttcacga atacagttgg attcactacg
     2341 ccgggattca aaaggccaac ctcgaggcct ggcccaagta ccgccaggcg ggtgtggagg
     2401 tcatccgcct gtctaacgag gacgtgcgca agttccgccg tctggccatc cccatctggt
     2461 tcaagtgggc caagatggac aagtactcgc gggaggcctt cgccagccag ctggagtaca
     2521 tgaagggcat cggctacgtt acggacgagg agctcaaggg gctttccctt gcggccgcag
     2581 aacaaaaact catctcagaa gaggatctga atggggccgc aagcgagaag gacgagctgt
     2641 agtctagaag ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     2701 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     2761 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2821 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatggc
     2881 ccgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     2941 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3001 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3061 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     3121 gctttacggc acctctcccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     3181 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3241 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     3301 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3361 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3421 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3481 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3541 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3601 ccctaggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3661 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctccccc
     3721 cccgggaggt ccacaatggt tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3781 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3841 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3901 gccctgaatg aactccagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3961 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4021 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4081 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4141 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4201 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4261 gcgcgtatgc ccgacggcga ggatctcgtc gtgactcatg gcgatgcctg cttgccgaat
     4321 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4381 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4441 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4501 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     4561 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4621 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4681 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     4741 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4801 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgatcttt ccgcttcctc
     4861 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4921 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4981 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5041 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5101 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5161 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5221 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5281 tatgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5341 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5401 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5461 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5521 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5581 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5641 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5701 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5761 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5821 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5881 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     5941 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6001 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6061 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6121 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6181 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6241 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6301 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6361 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6421 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6481 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcggg cacccaactg
     6541 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6601 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6661 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6721 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     6781 cgtcagatcg acggatcggg agatcg
//
