LOCUS       sequence-360537-        4843 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C. elegans plasmid encoding pH-Countdown with three introns.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4843)
  AUTHORS   Seidel ZP, Wang JCK, Riegler J, York AG, Ingaramo M
  TITLE     Relaxation Sensors
  JOURNAL   (2021), doi: 10.5281/zenodo.5810930
REFERENCE   2  (bases 1 to 4843)
  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: "(2021), 
            doi: 10.5281/zenodo.5810930"
COMMENT     Sequence Label: pUC57-Peft-pHCountdown-unc54
FEATURES             Location/Qualifiers
     source          1..4843
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        416..1015
                     /gene="eft-3"
                     /label=eft-3 promoter
                     /note="promoter for the C. elegans elongation factor 
                     1-alpha gene (Mitrovich and Anderson, 2000)"
     primer_bind     complement(2667..2683)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2667..2683)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2680..2702)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2691..2707
                     /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(2715..2721,2722..2739,2740..2745))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2760..2781
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2898..2915)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3069..3657)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3149..3168)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3828..4643)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3828..4643)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4551..4570
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(4644..4748)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4816..4834
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gacgcgtatt gggatgcacc
      421 tttggtcttt tattgtcaac ttccattggt tcttccattg tttctgttaa attaatgaat
      481 ttttcataaa ataaagacat tatacaatat aaaaatgaag aatttattga aaataaactg
      541 ccagagagaa aaagtatgca acactcccgc cgagagtgtt tgaaatggtg tacggtacat
      601 tttcgtgcta ggagttagat gtgcaggcag caacgagagg gggagagatt tttttgggcc
      661 ttgtgaaatt aacgtgagtt ttctggtcat ctgactaatc atgttggttt tttgttggtt
      721 tattttgttt ttatctttgt ttttatccag attaggaaat ttaaatttta tgaatttata
      781 atgaggtcaa acattcagtc ccagcgtttt tcctgttctc actgtttagt cgaattttta
      841 ttttaggctt tcaacaaatg ttctaactgt cttatttgtg acctcacttt ttatattttt
      901 ttaattttta aaaatattag aagtttctag gataattttt tcgactttta ttctctctac
      961 cgtccgcact cttcttactt ttaaattaaa ttgttttttt ttcagttggg aaacactttg
     1021 ctcactccgt agcagcctta attaaaaaat ggtttctaag ggagaagaaa acaacatggc
     1081 tgttatcaag ccagatatga agattaagct tcgtatggag ggatccgtta acggacaccg
     1141 cttccgcatc gagggagtcg gactcggaaa gccacttgaa ggaaagcaat ccatggatct
     1201 caaggttaag gtaagtttaa acatatatat actaactaac cctgattatt taaattttca
     1261 ggagggagga ccacttccat tcgcttacga catccttact atggctttct gctacggaaa
     1321 ccgtgttttc gctaagtacc cagagaacat cgttgattac ttcaagcaat ctttcccaga
     1381 aggatactcc tgggagcgcc aaatgatcta cgaggatgga ggaatctgcg ttgctaccaa
     1441 cgatatcacc cttgacggag attgcatgat ctctgagatc cgcttcaagg taagtttaaa
     1501 cagttcggta ctaactaacc atacatattt aaattttcag ggagttaatt tcccagctaa
     1561 cggaccagtc ttccaaaagc gcaccgttaa gtgggagctt tctcacgaga agctttacgc
     1621 ccgtgacgga cttctttact ccgacggaaa ctacgccctt tctctcgagg gaggaggaca
     1681 ctaccgctgc gataacaagg taagtttaaa catgatttta ctaactaact aatctgattt
     1741 aaattttcag accacctaca aggccaagaa ggtcgtccaa cttccagatt accactgggt
     1801 tactcacagt atcgttatca agtctcacga taaggactac tccaacgtca accttcacga
     1861 gcacgctgag gctcactccg agcttccacg tcaagctatg gatgagctct acaagtaact
     1921 cgagcccggg taagtccaat tactcttcaa catccctaca tgctctttct ccctgtgctc
     1981 ccacccccta tttttgttat tatcaaaaaa cttctcttaa tttctttgtt ttttagcttc
     2041 ttttaagtca cctctaacaa tgaaattgtg tagattcaaa aatagaatta attcgtaata
     2101 aaaagtcgaa aaaaattgtg ctccctcccc ccattaataa taattctatc ccaaaatcta
     2161 cacaatgttc tgtgtacact tcttatgttt tttacttctg ataaattttt ttgaaacatc
     2221 atagaaaaaa ccgcacacaa aataccttat catatgttac gtttcagttt atgaccgcaa
     2281 tttttatttc ttcgcacgtc tgggcctctc atgacgtcaa atcatgctca tcgtgaaaaa
     2341 gttttggagt atttttggaa tttttcaatc aagtgaaagt ttatgaaatt aattttcctg
     2401 cttttgcttt ttggggtttc ccctattgtt tgtcaagatt tcgaggacgg cgtttttctt
     2461 gctaaaatca caagtattga tgagcacgat gcaagaaaga tcggaagaag gtttgggttt
     2521 gaggctcagt ggaaggtgag tagaagttga taatttgaaa gtggagtagt gtctatgggg
     2581 tttttgcctt aaatgacaga atacattccc aatataccaa acataactgt ttatcccaat
     2641 ggcgcgccga gcttggctcg agcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     2701 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     2761 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     2821 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     2881 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     2941 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3001 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3061 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3121 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3181 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3241 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3301 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3361 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3421 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3481 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3541 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3601 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3661 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3721 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3781 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta gaaaaactca
     3841 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
     3901 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     3961 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     4021 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
     4081 aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
     4141 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga
     4201 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     4261 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     4321 tggaatgctg ttttcccagg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     4381 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     4441 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     4501 tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
     4561 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac
     4621 gtttcccgtt gaatatggct catactcttc ctttttcaat attattgaag catttatcag
     4681 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     4741 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     4801 acattaacct ataaaaatag gcgtatcacg aggccctttc gtc
//
