LOCUS       sequence-453770-        5687 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of green fluorescent sodium ion indicator 
            HaloGFP-Na2.4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5687)
  AUTHORS   Takeuchi S, Imai S, Terai T, Campbell RE
  TITLE     A chemigenetic indicator based on a synthetic chelator and a green 
            fluorescent protein for imaging of intracellular sodium ions.
  JOURNAL   RSC Chem Biol. 2024 Dec 3. doi: 10.1039/d4cb00256c.
  PUBMED    39678364
REFERENCE   2  (bases 1 to 5687)
  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: "RSC Chem 
            Biol. 2024 Dec 3. doi: 10.1039/d4cb00256c."
COMMENT     Sequence Label: pBAD-HaloGFP-Na2.4
FEATURES             Location/Qualifiers
     source          1..5687
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     30..47
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    62..202
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(128..147)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     288..310
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(466..1344)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1371..1655
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1578..1597
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1701..1718
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1722..1754
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(1758..1766,1767..1781)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(2625..2646)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2886..2905)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(3458..3475)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3628..3714
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3806..3833
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3852..3943
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3944..4012,4013..4804)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4162..4181)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4975..5563
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5464..5483
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
       61 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca
      121 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
      181 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
      241 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
      301 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagcagatca attcgcgcgc
      361 gaaggcgaag cggcatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac
      421 cctatgctac tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac
      481 ggctacatca ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt
      541 gcatttttta aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac
      601 ggtggcgata ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc
      661 ctcgcgccag cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga
      721 cggcgacaag caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg
      781 atcgctgatg tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc
      841 gttaatcgct tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc
      901 cgaatagcgc ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg
      961 cggctggtgc gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt
     1021 aagccattca tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg
     1081 agcctccgga tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc
     1141 cggtcggcaa acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag
     1201 attgagaata taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt
     1261 ggcctcaatc ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag
     1321 gggatcattt tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa
     1381 ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac
     1441 caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac
     1501 aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca
     1561 cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg
     1621 acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc taacaggagg
     1681 aattaaccat ggggggttct catcatcatc atcatcatgg tatggctagc atgactggtg
     1741 gacagcaaat gggtcgggat ctgtacgacg atgacgataa ggatccgagc tcgagaatgg
     1801 cagaaatcgg tactggcttt ccattcgacc cccattatgt ggaagtcctg ggcgagcgca
     1861 tgcactacgt cgatgttggt ccgcgcgatg gcacccctgt gctgttcctg cacggtaacc
     1921 cgacctcctc ctacgtgtgg cgcaacatca tcccgcatgt tgcaccgacc catcgctgca
     1981 ttgctccaga cctgatcggt atgggcaaat ccgacaaacc agacctgggt tatttcttcg
     2041 acgaccacgt ccgcttcatg gatgccttca tcgaagccct gggtctggaa gaggtcgtcc
     2101 tggtcattca cgactggggc tccgctctgg gcttccactg ggccaagcgc aatccagagc
     2161 gcgtcaaagg tattgcattt atggagttca tccgccctat cccgacctgg gacgaatggc
     2221 cagaatttgc ccgcgagacc ttccaggcct tccgcaccac cgacgtcggc cgcaagctga
     2281 tcatcggtga aaacgtctat atcaaggccg acaagcagaa gaacggcatc aaggcgaact
     2341 tcgagatccg ccacaacacc gaggacggcg gcgtgcagct cgcctaccac taccagcaga
     2401 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcgtccagt
     2461 ccaaactttc gaaagacccc aacgagaagc gcgatcacat ggtcctgctg gagctcgtga
     2521 ccgccgccgg gatcactctc ggcatggacg agctgtacga gggcggtacc ggagggagca
     2581 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     2641 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg tgagggcgat gccacctacg
     2701 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     2761 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     2821 agcacgactt cttcaagtcc gccatgcccg aaggctacat ccaggagcgc accatcttct
     2881 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     2941 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcata
     3001 agctggagta caacctgtcg aattttttta tcgagggtac gctgccgatg ggtgtcgtcc
     3061 gcccgctgac tgaggtcgag atggaccatt accgcgagcc gttcctgaat cctgttgacc
     3121 gcgagccact gtggcgcttc ccaaacgagc tgccaatcgc cggtgagcca gcgaacatcg
     3181 tcgcgctggt cgaagaatac atggactggc tgcaccagtc ccctgtcccg aagctgctgt
     3241 tctggggcac cccaggcgtt ctgatcccac cggccgaagc cgctcgcctg gccaaaagcc
     3301 tgcctaactg caaggctgtg gacatcggcc cgggtctgaa tctgctgcaa gaagacaacc
     3361 cggacctgat cggcagcgag atcgcgcgct ggctgtcgac gcacgagatt cccggctgaa
     3421 agcttggctg ttttggcgga tgagagaaga ttttcagcct gatacagatt aaatcagaac
     3481 gcagaagcgg tctgataaaa cagaatttgc ctggcggcag tagcgcggtg gtcccacctg
     3541 accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc
     3601 atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     3661 gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg
     3721 ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca
     3781 taaactgcca ggcatcaaat taagcagaag gccatcctga cggatggcct ttttgcgttt
     3841 ctacaaactc ttttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3901 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     3961 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4021 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4081 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4141 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     4201 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4261 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4321 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4381 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4441 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4501 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4561 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4621 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4681 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4741 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4801 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     4861 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4921 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4981 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa caaccaccgc taccagcggt
     5041 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5101 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     5161 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5221 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5281 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5341 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5401 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5461 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5521 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5581 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     5641 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgata
//
