LOCUS       sequence-496179-        6588 bp    DNA     circular SYN 05-JAN-2026
DEFINITION  Transient expression of the indicator protein CLOMELEON for 
            monitoring chloride concentrations in the cytoplasm of plant cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6588)
  AUTHORS   Lorenzen I, Aberle T, Plieth C
  TITLE     Salt stress-induced chloride flux: a study using transgenic 
            Arabidopsis expressing a fluorescent anion probe.
  JOURNAL   Plant J. 2004 May;38(3):539-44. doi: 
            10.1111/j.0960-7412.2004.02053.x.
  PUBMED    15086798
REFERENCE   2  (bases 1 to 6588)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 5, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1111/j.0960-7412.2004.02053"; journalName: "Plant J"; date: 
            "2004-05"; volume: "38"; issue: "3"; pages: "539-44"
COMMENT     Sequence Label: pART7::CLOMELEON
FEATURES             Location/Qualifiers
     source          1..6588
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(19..36)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(19..35)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(28..50)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(176..631)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(313..334)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     525..544
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        658..762
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(763..831,832..1623)
                     /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     complement(981..1000)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1794..2382
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2283..2302
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2536..2553
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2670..2691
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2706..2711,2712..2729,2730..2736)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2744..2760
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2749..2771
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2768..2784
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2768..2784
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2814..2833
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2814..2832
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        3850..4195
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     4138..4158
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     RBS             4213..4224
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4243..4260
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(4285..4287,4288..4290,4291..5001)
                     /codon_start=1
                     /product="cyan variant of GFP"
                     /label=CFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             5026..5046
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(5074..5076,5077..5079,5080..5790)
                     /codon_start=1
                     /product="enhanced yellow variant of GFP"
                     /label=Topaz YFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMRQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     terminator      5812..6519
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(6560..6578)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6561..6578)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 tgaccaagtc agcttggcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
       61 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
      121 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggaaatt
      181 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
      241 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
      301 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
      361 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
      421 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
      481 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
      541 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
      601 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
      661 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
      721 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
      781 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
      841 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
      901 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
      961 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     1021 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     1081 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     1141 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     1201 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     1261 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     1321 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     1381 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     1441 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     1501 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     1561 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     1621 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     1681 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     1741 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     1801 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     1861 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     1921 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     1981 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2041 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2101 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2161 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     2221 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     2281 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     2341 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     2401 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     2461 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     2521 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     2581 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     2641 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     2701 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     2761 tttcacacag gaaacagcta tgacatgatt acgaatttgg ccaagtcggc ctctaatacg
     2821 actcactata gggagctcgt cgagcggccg ctcgacgaat taattccaat cccacaaaaa
     2881 tctgagctta acagcacagt tgctcctctc agagcagaat cgggtattca acaccctcat
     2941 atcaactact acgttgtgta taacggtcca catgccggta tatacgatga ctggggttgt
     3001 acaaaggcgg caacaaacgg cgttcccgga gttgcacaca agaaatttgc cactattaca
     3061 gaggcaagag cagcagctga cgcgtacaca acaagtcagc aaacagacag gttgaacttc
     3121 atccccaaag gagaagctca actcaagccc aagagctttg ctaaggccct aacaagccca
     3181 ccaaagcaaa aagcccactg gctcacgcta ggaaccaaaa ggcccagcag tgatccagcc
     3241 ccaaaagaga tctcctttgc cccggagatt acaatggacg atttcctcta tctttacgat
     3301 ctaggaagga agttcgaagg tgaaggtgac gacactatgt tcaccactga taatgagaag
     3361 gttagcctct tcaatttcag aaagaatgct gacccacaga tggttagaga ggcctacgca
     3421 gcaggtctca tcaagacgat ctacccgagt aacaatctcc aggagatcaa ataccttccc
     3481 aagaaggtta aagatgcagt caaaagattc aggactaatt gcatcaagaa cacagagaaa
     3541 gacatatttc tcaagatcag aagtactatt ccagtatgga cgattcaagg cttgcttcat
     3601 aaaccaaggc aagtaataga gattggagtc tctaaaaagg tagttcctac tgaatctaag
     3661 gccatgcatg gagtctaaga ttcaaatcga ggatctaaca gaactcgccg tgaagactgg
     3721 cgaacagttc atacagagtc ttttacgact caatgacaag aagaaaatct tcgtcaacat
     3781 ggtggagcac gacactctgg tctactccaa aaatgtcaaa gatacagtct cagaagacca
     3841 aagggctatt gagacttttc aacaaaggat aatttcggga aacctcctcg gattccattg
     3901 cccagctatc tgtcacttca tcgaaaggac agtagaaaag gaaggtggct cctacaaatg
     3961 ccatcattgc gataaaggaa aggctatcat tcaagatctc tctgccgaca gtggtcccaa
     4021 agatggaccc ccacccacga ggagcatcgt ggaaaaagaa gacgttccaa ccacgtcttc
     4081 aaagcaagtg gattgatgtg acatctccac tgacgtaagg gatgacgcac aatcccacta
     4141 tccttcgcaa gacccttcct ctatataagg aagttcattt catttggaga ggacacgctc
     4201 gaggaattca ttaaagagga gaaattaact atgagaggat cgcatcacca tcaccatcac
     4261 ggatccgcat gcgagctcgg taccatggtg agcaagggcg aggagctgtt caccggggtg
     4321 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaggttcag cgtgtccggc
     4381 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     4441 aagctgcccg tgccctggcc caccctcgtg accaccctga cctggggcgt gcagtgcttc
     4501 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     4561 tacgtccagg agcgtaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     4621 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     4681 gaggacggca acatcctggg gcacaagctg gagtacaact acatcagcca caacgtctat
     4741 atcaccgccg acaagcagaa gaacggcatc aaggcccact tcaagatccg ccacaacatc
     4801 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     4861 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     4921 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     4981 ggcatggacg agctgtacaa gaagctcacc ggtagtggca gtggtgagaa tttgtacttc
     5041 caaggaggcg gcagcggagg cactagtagt accatggtga gcaagggcga ggagctgttc
     5101 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     5161 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     5221 accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcgtg
     5281 cagtgcttcg cccgctaccc cgaccacatg cgccagcacg acttcttcaa gtccgccatg
     5341 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     5401 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     5461 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     5521 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     5581 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     5641 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc
     5701 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     5761 atcactctcg gcatggacga gctgtacaag aagcttggat cctctagagt cctgctttaa
     5821 tgagatatgc gagacgccta tgatcgcatg atatttgctt tcaattctgt tgtgcacgtt
     5881 gtaaaaaacc tgagcatgtg tagctcagat ccttaccgcc ggtttcggtt cattctaatg
     5941 aatatatcac ccgttactat cgtattttta tgaataatat tctccgttca atttactgat
     6001 tgtaccctac tacttatatg tacaatatta aaatgaaaac aatatattgt gctgaatagg
     6061 tttatagcga catctatgat agagcgccac aataacaaac aattgcgttt tattattaca
     6121 aatccaattt taaaaaaagc ggcagaaccg gtcaaaccta aaagactgat tacataaatc
     6181 ttattcaaat ttcaaaagtg ccccaggggc tagtatctac gacacaccga gcggcgaact
     6241 aataacgctc actgaaggga actccggttc cccgccggcg cgcatgggtg agattccttg
     6301 aagttgagta ttggccgtcc gctctaccga aagttacggg caccattcaa cccggtccag
     6361 cacggcggcc gggtaaccga cttgctgccc cgagaattat gcagcatttt tttggtgtat
     6421 gtgggcccca aatgaagtgc aggtcaaacc ttgacagtga cgacaaatcg ttgggcgggt
     6481 ccagggcgaa ttttgcgaca acatgtcgag gctcagcagg acctgcaggc atgcaagcta
     6541 gcttactagt gatgcatatt ctatagtgtc acctaaatct gcggccgc
//
