LOCUS       sequence-378876-        6152 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Turn-on fluorescent sensor for chloride.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6152)
  AUTHORS   Tutol JN, Ong WSY, Phelps SM, Peng W, Goenawan H, Dodani SC
  TITLE     Engineering the ChlorON Series: Turn-On Fluorescent Protein Sensors 
            for Imaging Labile Chloride in Living Cells.
  JOURNAL   ACS Cent Sci. 2023 Dec 18;10(1):77-86. doi: 
            10.1021/acscentsci.3c01088. eCollection 2024 Jan 24.
  PUBMED    38292617
REFERENCE   2  (bases 1 to 6152)
  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/acscentsci.3c01088"; journalName: "ACS Cent Sci"; date: 
            "2023-12-18- 18"; volume: "10"; issue: "1"; pages: "77-86"
COMMENT     Sequence Label: ChlorON-1
FEATURES             Location/Qualifiers
     source          1..6152
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(49..68)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        240..619
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        620..823
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     774..794
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     868..887
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        868..886
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             931..948
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      1222..1231
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1748..1765)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1754..1978
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2024..2452
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2111..2130)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2321..2342
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2461..2481)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2466..2795
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2646..2781
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2708..2727
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2862..3656
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2916..2935)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3526..3545
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3830..3951
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3867..3886)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3921..3940
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4000..4016)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4000..4016)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4013..4035)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4024..4040
                     /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)."
     promoter        complement(join(4048..4054,4055..4072,4073..4078))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4093..4114
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4231..4248)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4402..4990)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4482..4501)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5161..5952,5953..6021))
                     /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     5784..5803
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6022..6126)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc
       61 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
      121 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
      181 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg
      241 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
      301 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      361 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      421 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      481 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      541 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      601 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      661 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      721 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      781 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga
      841 acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg
      901 ctagcgttta aacttaagct tgccaccatg catcatcacc atcaccatgg taccgagctc
      961 ggatccatgg tgtccaaggg cgaggaggac aatatggcct ctctgccagc cacccacgag
     1021 ctgcacatct tcggctctat caacggcgtg gactttgata tggtgggaca gggaaccgga
     1081 aacccaaatg acggctacga ggagctgaat ctgaagtcta caaagggcga tctgcagttc
     1141 agcccttgga ttctggtgcc acacatcggc tatggctttc accagtatct gccctaccct
     1201 gacggcatgt ctcctttcca ggccgccatg gtggatggca gcggctacca ggtgcacagg
     1261 acaatgcagt ttgaggacgg cgcctccctg accgtgaact accgctatac atacgagggc
     1321 tctcacatca agggagaggc acaggtgaag ggaaccggat tcccagcaga tggacccgtg
     1381 atgaccaaca gcctgacagc agcagactgg tgccggtcca agtggacata tcccaatgat
     1441 aagaccatca tcagcacctt caagtggtcc tataccacag gcaacggcaa gcggtacaga
     1501 agcaccgccc ggaccacata tacatttgcc aagcccatgg ccgccaacta tctgaagaat
     1561 cagcctatgt acgtgttcct gaagaccgag ctgaagcact ccaagacaga gctgaatttc
     1621 aaggagtggc agaaggcctt taccgacgtg atgggcatgg atgagctgta caagtgagaa
     1681 ttctgcagat atccagcaca gtggcggccg ctcgagtcta gagggcccgt ttaaacccgc
     1741 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     1801 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     1861 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     1921 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     1981 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     2041 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     2101 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     2161 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     2221 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     2281 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     2341 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     2401 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     2461 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2521 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     2581 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     2641 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     2701 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     2761 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     2821 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     2881 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     2941 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3001 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     3061 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     3121 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     3181 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     3241 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     3301 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     3361 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     3421 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     3481 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     3541 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     3601 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     3661 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     3721 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     3781 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     3841 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3901 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     3961 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4021 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4081 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4141 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4201 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4261 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4321 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4381 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4441 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4501 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4561 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4621 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4681 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4741 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4801 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4861 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4921 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4981 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5041 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5101 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5161 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5221 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5281 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5341 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5401 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5461 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5521 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5581 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5641 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5701 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5761 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5821 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5881 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5941 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6001 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6061 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6121 tccgcgcaca tttccccgaa aagtgccacc tg
//
