LOCUS       sequence-355806-        4752 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luminal ER expression of ratiometric pHLuorin2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4752)
  AUTHORS   Linders PTA, Ioannidis M, Ter Beest M, van den Bogaart G
  TITLE     Fluorescence Lifetime Imaging of pH along the Secretory Pathway.
  JOURNAL   ACS Chem Biol. 2022 Jan 21;17(1):240-251. doi: 
            10.1021/acschembio.1c00907. Epub 2022 Jan 10.
  PUBMED    35000377
REFERENCE   2  (bases 1 to 4752)
  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: 
            "10.1021/acschembio.1c00907"; journalName: "ACS Chem Biol"; date: 
            "2022-01-21- 21"; volume: "17"; issue: "1"; pages: "240-251"
COMMENT     Sequence Label: ER-RpHLuorin2
FEATURES             Location/Qualifiers
     source          1..4752
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(195..214)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    239..286
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      615..1203
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1104..1123
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1378..1681
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1682..1885
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1836..1856
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(2295..2314)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2878..2999
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2915..2934)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2969..2988
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3006..3461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3143..3164)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3355..3374
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3488..3592
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3594..3785
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3617..3637)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3782..3916
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(3965..4752,1..7)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             join(3965..4752,1..7)
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4019..4038)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4629..4648
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
       61 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
      121 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
      181 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
      241 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
      301 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
      361 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
      421 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
      481 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
      541 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
      601 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
      661 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
      721 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
      781 ttaggccacc acttcaagaa ctctgtatca ccgcctacat acctcgctct gctaatcctg
      841 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
      901 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
      961 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     1021 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     1081 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     1141 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     1201 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     1261 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag
     1321 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     1381 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     1441 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     1501 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     1561 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     1621 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     1681 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     1741 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     1801 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     1861 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agcatgctgc
     1921 tatccgtgcc gttgctgctc ggcctcctcg gcctggccgt cgccgaccgg tcaggcggca
     1981 gcggcggcag cgctagcaag ggcgaggagc tgttcaccgg cgtggtgccc atcctggtgg
     2041 agctggacgg cgacgtgaac ggccacaagt tcagcgtgag cggcgagggc gagggcgacg
     2101 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     2161 ggcccaccct ggtgaccacc ctgagctacg gcgtgcagtg cttcagccgc taccccgacc
     2221 acatgaagca gcacgacttc ttcaagagcg ccatgcccga gggctacgtg caggagcgca
     2281 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     2341 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     2401 tgggccacaa gctggagtac aactacaacg agcacctggt ctacatcatg gccgacaagc
     2461 agaagaacgg caccaaggcc atcttccagg tgcaccacaa catcgaggac ggcggcgtgc
     2521 agctggccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     2581 acaaccacta cctgcacacc cagagcgccc tgagcaagga ccccaacgag aagcgcgacc
     2641 acatggtgct gctggagttc gtgaccgccg ccggcatcac ccacggcatg gacgagctgt
     2701 acaaggacga gctgtaagaa ttctgcagtc gacggtaccg cgggcccggg atccaccgga
     2761 tctagataac tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     2821 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     2881 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     2941 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     3001 ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     3061 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     3121 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     3181 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     3241 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     3301 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     3361 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     3421 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     3481 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3541 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     3601 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3661 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3721 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3781 gtcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3841 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     3901 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     3961 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     4021 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     4081 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4141 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4201 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4261 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4321 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4381 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4441 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     4501 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4561 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4621 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4681 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4741 tcttgacgag tt
//
