LOCUS       sequence-355808-        4980 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cis-/medial-Golgi expression of ratiometric pHLuorin2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4980)
  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 4980)
  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: MGAT2-RpHLuorin2
FEATURES             Location/Qualifiers
     source          1..4980
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        51..354
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        355..558
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     509..529
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(1222..1241)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    1758..1879
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1795..1814)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1849..1868
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1886..2341)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2023..2044)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2235..2254
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2368..2472
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2474..2831
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2474..2665
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2497..2517)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2682..2817
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2866..3660
                     /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             2866..3660
                     /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(2920..2939)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3530..3549
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3848..3867)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3892..3939
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4268..4856
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4757..4776
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
       61 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      121 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      181 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      241 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      301 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      361 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      421 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      481 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
      541 gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta ccggactcag
      601 atctcgagct caagcttcga attcatgcgc ttccgcatct acaagcgcaa ggtgctgatc
      661 ctgaccctgg tggtggccgc ctgcggcttc gtgctgtgga gcagcaacgg ccgccagcgc
      721 aagaacgagg ccctggcccc ccccctgctg gacgccgagc ccgcccgcgg cgccggcggc
      781 cgcggcggcg accaccccag cgtggccgtg ggcatccgcc gcgtgagcaa cgtgagcgcc
      841 gccagcctgg tgcccgccgt gccccagccc gaggccgaca acctgaccct gttggatcca
      901 ggcggcagcg gcggcagcgc tagcaagggc gaggagctgt tcaccggcgt ggtgcccatc
      961 ctggtggagc tggacggcga cgtgaacggc cacaagttca gcgtgagcgg cgagggcgag
     1021 ggcgacgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     1081 gtgccctggc ccaccctggt gaccaccctg agctacggcg tgcagtgctt cagccgctac
     1141 cccgaccaca tgaagcagca cgacttcttc aagagcgcca tgcccgaggg ctacgtgcag
     1201 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1261 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1321 aacatcctgg gccacaagct ggagtacaac tacaacgagc acctggtcta catcatggcc
     1381 gacaagcaga agaacggcac caaggccatc ttccaggtgc accacaacat cgaggacggc
     1441 ggcgtgcagc tggccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1501 ctgcccgaca accactacct gcacacccag agcgccctga gcaaggaccc caacgagaag
     1561 cgcgaccaca tggtgctgct ggagttcgtg accgccgccg gcatcaccca cggcatggac
     1621 gagctgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag
     1681 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     1741 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     1801 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     1861 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     1921 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     1981 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     2041 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     2101 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     2161 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     2221 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     2281 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     2341 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2401 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     2461 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     2521 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2581 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2641 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2701 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     2761 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     2821 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     2881 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     2941 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3001 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     3061 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3121 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3181 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3241 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3301 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3361 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     3421 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     3481 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     3541 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     3601 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     3661 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     3721 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     3781 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     3841 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     3901 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     3961 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     4021 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     4081 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     4141 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4201 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4261 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4321 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4381 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4441 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4501 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4561 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4621 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4681 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4741 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4801 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4861 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4921 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa
//
