LOCUS       sequence-149375-        4433 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses YhjH under a T7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4433)
  AUTHORS   Wang XC, Wilson SC, Hammond MC
  TITLE     Next-generation RNA-based fluorescent biosensors enable anaerobic 
            detection of cyclic di-GMP.
  JOURNAL   Nucleic Acids Res. 2016 Jul 5. pii: gkw580.
  PUBMED    27382070
REFERENCE   2  (bases 1 to 4433)
  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: "Nucleic 
            Acids Res. 2016 Jul 5. pii: gkw580."
COMMENT     Sequence Label: pCOLA-T7-YhjH
FEATURES             Location/Qualifiers
     source          1..4433
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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)."
     CDS             83..100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        214..232
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    233..257
                     /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)."
     CDS             1080..1124
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1162..1180)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1176..1223
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1453..2268)
                     /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(1453..2268)
                     /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     2176..2195
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2269..2360)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2378..3013)
                     /direction=LEFT
                     /label=ColA ori
                     /note="Plasmids containing the ColA origin of replication 
                     can be propagated in E. coli cells that contain additional 
                     plasmids with compatible origins."
     primer_bind     complement(2523..2542)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3211..4293)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4255..4274
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4294..4371)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(4417..4433,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cgaattcgag
      121 ctcggcgcgc ctgcaggtcg acaagcttgc ggccgcataa tgcttaagtc gaacagaaag
      181 taatcgtatt gtacacggcc gcataatcga aattaatacg actcactata ggggaattgt
      241 gagcggataa caattcccca tcttagtata ttagttaagt ataagaagga gatatacata
      301 tgataaggca ggttatccag cgaataagca accctgaagc aagcatcgag agcttgcagg
      361 aacggcgttt ttggttgcag tgtgagcgtg cttacacctg gcagccgatc tatcaaacat
      421 gcgggcggtt aatggccgtg gagctattaa cggtggtcac gcatcccttg aacccttcgc
      481 aacgcctgcc gccggatcgc tattttactg aaatcaccgt cagccatcgg atggaggttg
      541 tgaaagagca gattgatttg ctggcgcaaa aagccgactt ctttatagag cacggcctgc
      601 tggcatcggt caatattgat ggccctacgc tcatcgccct gcgtcagcaa ccaaaaatcc
      661 tgcgccagat tgagcgtctt ccctggctgc gtttcgaact ggtggagcat atccgtctgc
      721 cgaaagattc aacctttgcc tcgatgtgtg aatttggccc gctgtggctg gatgattttg
      781 gtaccgggat ggcaaatttc tctgcgctaa gtgaagtgcg ttatgactac atcaaaatcg
      841 cgcgagaact gtttgtgatg ctgcgtcagt cgccggaagg acgcacactc ttttctcagc
      901 ttttacatct aatgaatcgc tattgtcgcg gggtgattgt cgagggcgta gaaacgccgg
      961 aagagtggcg tgatgttcag aactcgcccg cattcgccgc acaaggctgg tttctttcac
     1021 gcccggcacc gatagaaacg ctgaatacgg cggttctggc gctataactc gagtctggta
     1081 aagaaaccgc tgctgcgaaa tttgaacgcc agcacatgga ctcgtctact agcgcagctt
     1141 aattaaccta ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
     1201 aacgggtctt gaggggtttt ttgctgaaac ctcaggcatt tgagaagcac acggtcacac
     1261 tgcttccggt agtcaataaa ccggtaaacc agcaatagac ataagcggct atttaacgac
     1321 cctgccctga accgacgaca agctgacgac cgggtctccg caagtggcac ttttcgggga
     1381 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     1441 atgaattaat tcttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc
     1501 aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc
     1561 gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac
     1621 atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc
     1681 atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg
     1741 ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt
     1801 cattcgtgat tgcgcctgag cgagacgaaa tacgcggtcg ctgttaaaag gacaattaca
     1861 aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc
     1921 tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag
     1981 taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc
     2041 cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc
     2101 atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc
     2161 tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga
     2221 atttaatcgc ggcctagagc aagacgtttc ccgttgaata tggctcatac tcttcctttt
     2281 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     2341 tatttagaaa aataaacaaa taggcatgct agcgcagaaa cgtcctagaa gatgccagga
     2401 ggatacttag cagagagaca ataaggccgg agcgaagccg tttttccata ggctccgccc
     2461 ccctgacgaa catcacgaaa tctgacgctc aaatcagtgg tggcgaaacc cgacaggact
     2521 ataaagatac caggcgtttc cccctgatgg ctccctcttg cgctctcctg ttcccgtcct
     2581 gcggcgtccg tgttgtggtg gaggctttac ccaaatcacc acgtcccgtt ccgtgtagac
     2641 agttcgctcc aagctgggct gtgtgcaaga accccccgtt cagcccgact gctgcgcctt
     2701 atccggtaac tatcatcttg agtccaaccc ggaaagacac gacaaaacgc cactggcagc
     2761 agccattggt aactgagaat tagtggattt agatatcgag agtcttgaag tggtggccta
     2821 acagaggcta cactgaaagg acagtatttg gtatctgcgc tccactaaag ccagttacca
     2881 ggttaagcag ttccccaact gacttaacct tcgatcaaac cgcctcccca ggcggttttt
     2941 tcgtttacag agcaggagat tacgacgatc gtaaaaggat ctcaagaaga tcctttacgg
     3001 attcccgaca ccatcactct agatttcagt gcaatttatc tcttcaaatg tagcacctga
     3061 agtcagcccc atacgatata agttgtaatt ctcatgttag tcatgccccg cgcccaccgg
     3121 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
     3181 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     3241 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3301 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     3361 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     3421 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
     3481 cactaccgag atgtccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     3541 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     3601 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     3661 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     3721 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     3781 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     3841 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     3901 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     3961 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     4021 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     4081 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     4141 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     4201 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     4261 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     4321 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
     4381 gacgctctcc cttatgcgac tcctgcatta ggaaattaat acgactcact ata
//
