LOCUS       sequence-192672-        4772 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  drives expression of gnd using Ptrc promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4772)
  AUTHORS   Puszynska AM, O'Shea EK
  TITLE     Switching of metabolic programs in response to light availability is
            an essential function of the cyanobacterial circadian output 
            pathway.
  JOURNAL   Elife. 2017 Apr 21;6. pii: e23210. doi: 10.7554/eLife.23210.
  PUBMED    28430105
REFERENCE   2  (bases 1 to 4772)
  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: "Elife."; 
            date: "2017-04-21"; volume: "6. pii: e23210. doi"; pages: " 
            10.7554/eLife.23210"
COMMENT     Sequence Label: Ptrc::gnd
FEATURES             Location/Qualifiers
     source          1..4772
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(1034..1849)
                     /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(1034..1849)
                     /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     1757..1776
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1841..1860
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     2198..2220
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        2298..2375
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             2376..3458
                     /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     complement(2395..2414)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(3690..3695,3696..3712,3713..3719)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    3727..3743
                     /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     3732..3754
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 agccagccta agccgtctgt cgatcccagc agcgatagag atggaggagt tgatcgaaat
       61 agtgacgagc taattccagc tgagggaaag acttcaccga aagctgatcg ctccggcaat
      121 ccaaaacacg aacttgatga tcttcaatat tggcaactgc aggactgaga atgacgatcc
      181 agcgaccgat atgttgtcgt tgcgtttcca taagaaagac agatgattgt gcgattaaag
      241 tagcgacaac aaggctaaca aatcgttacc tagggcacga ttttgctctg aaccagagtc
      301 agtcagcaca acgctaaact tttgtagagc tttatcctcg gctcctaaac ggttgcgatc
      361 gccatgttga ttcagcctgc tgcaacgacg tctaccccag aggtggctct cgcaatcatc
      421 acccaaggtc aacagtggtt attgcagctg cgtgatgact tagattggat tctctatccg
      481 gggaagtggg gcttatttgg cggccattta gaaccggatg aaactcccct tgccgccctc
      541 tatcgcgaac tccaagaaga gattggctac tacccctccg ccgcaacccc actctggacg
      601 gaaacagaag aaagggtcac tcgctacctc ttccatgtgc ctttgacctg tgaactatcg
      661 actttgcggc tcggcgaagg gcaggatttc tgcctcgtcg atgccgccgc gatcgcccaa
      721 ggtcgagttt tctcgccccg tttacagcag gaacgagcga tcggtgcgcc ccaccagcag
      781 attttgctgc gcttccaagg ggagtattcg cacaagcttg cccccgtcat cagggcctga
      841 ccggtgcgat cgccgtcaag aatccacccc caggcattca atttttctct gatctgtaaa
      901 taaatccgcg atcgccgccg tgatccccgc aaagaaagtc taaagaggtt gccagagaac
      961 cccaaattgg caggctaacc tgaggtcatt cgttgtccta ctgccagtgt tacaaccaat
     1021 taaccaattc tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat
     1081 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac
     1141 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa
     1201 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac
     1261 catgagtgac gactgaatcc ggtgagaatg gcaaaagctt atgcatttct ttccagactt
     1321 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat
     1381 tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac
     1441 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
     1501 ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga
     1561 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
     1621 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc
     1681 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac
     1741 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
     1801 aatttaatcg cggcctcgag caagacgttt cccgttgaat atggctcata acaccccttg
     1861 tattactgtt tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg
     1921 caatgtaaca tcagagattt tgagacacaa cgtggctttg ttgaataaat cgaacttttg
     1981 ctgagttgaa ggatcagatc acgcatcttc ccgacaacgc agtgctagct cgagcccgcg
     2041 acatcttcct gctccagaag ctcgaaattc gagctcggta cccggggatc tgggccgcgg
     2101 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     2161 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     2221 ttttcaccgt catcaccgaa acgcgcgagg cagcagatca attcgcgcgc gaaggcgaag
     2281 cggcatgcat ttacgttgac accatcgaat ggtgcaaaac ctttcgcggt atggcatgat
     2341 agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg
     2401 tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc
     2461 acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg aattacattc
     2521 ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc gttgccacct
     2581 ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc
     2641 aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag
     2701 cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac tatccgctgg
     2761 atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg ttatttcttg
     2821 atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac ggtacgcgac
     2881 tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta gcgggcccat
     2941 taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc actcgcaatc
     3001 aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt tttcaacaaa
     3061 ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc aacgatcaga
     3121 tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt gcggatatct
     3181 cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg ttaaccacca
     3241 tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg caactctctc
     3301 agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa agaaaaacca
     3361 ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     3421 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtaagt
     3481 tagcgcgaat tgatctggtt tgacagctta tcatcgactg cacggtgcac caatgcttct
     3541 ggcgtcaggc agccatcgga agctgtggta tggctgtgca ggtcgtaaat cactgcataa
     3601 ttcgtgtcgc tcaaggcgca ctcccgttct ggataatgtt ttttgcgccg acatcataac
     3661 ggttctggca aatattctga aatgagctgt tgacaattaa tcatccggct cgtataatgt
     3721 gtggaattgt gagcggataa caatttcaca caggaaacag accatggaat tcatggcact
     3781 gcaacagttc ggcctcatcg ggctggcagt gatgggtgag aatctcgccc tgaacattga
     3841 gcgcaacggt ttttccttga cggtctataa ccgcacggcc gaaaagactg aagcgttcat
     3901 ggccgatcgc gcccaaggca agaacattgt gccggcttac agtctggaag actttgttgc
     3961 cagcttggaa cgtccgcgcc gcattttggt gatggttaaa gcgggcggcc cggtggatgc
     4021 cgtggtcgag cagctcaaac ccctgctaga tcccggtgac ttgatcatcg atgggggcaa
     4081 ctcactgttt accgacaccg agcgccgcgt caaagatcta gaagcgctag gtctgggttt
     4141 catgggcatg ggcgtcagtg gtggcgaaga gggggctttg aatggcccca gtttgatgcc
     4201 ggggggcacc caagcggcct acgaagccgt ggagccgatc gtgcgcagca ttgctgccca
     4261 agtcgacgat ggcccctgcg ttacctacat cggtcctggt ggctctgggc actacgtcaa
     4321 gatggtgcac aacggcattg aatatggcga tatgcagctg atcgccgaag cctatgacct
     4381 cctcaaatcg gtggcaggcc tcaatgccag tgaactgcat gatgtcttcg ccgcctggaa
     4441 caaaacgccg gaactcgact cgttcttgat tgaaatcacg gcggatattt tcaccaaagt
     4501 cgatgacttg ggtactggtc agcccttggt cgagctgatt ttagatgccg ctggtcaaaa
     4561 aggaaccggt cgctggacgg tggaaacggc actggaaatt ggcgttgcga ttccaaccat
     4621 cattgctgct gtcaacgccc gcatcctgtc ctcgatcaaa gccgagcgtc aggcagcttc
     4681 tgaaattctt tcaggaccga ttaccgagcc cttcagtggc gatcgccaag cctttatcga
     4741 cagtgtgcgc gatgcgctct actgctcgaa aa
//
