LOCUS       sequence-192670-        5330 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Replaces native promoter of glgP_gap1 with Ptrc promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5330)
  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 5330)
  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::glgP_gap1
FEATURES             Location/Qualifiers
     source          1..5330
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     1011..1030
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1249..1353)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1421..1439
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1556..1658
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1659..2318
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1725..1744)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     2759..2781
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        2859..2936
                     /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             2937..4019
                     /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(2956..2975)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4251..4256,4257..4273,4274..4280)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    4288..4304
                     /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     4293..4315
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 tgataccaaa ctcctccgcc tgaatcgtgg cattgaccac ccatttgaca acgtcaaacc
       61 aaggcgagtc attgttgagg gtggctgggc tcagcggctc tttggagatc actgcatcca
      121 gcaattggtg ctgatcggca tcggacaagg tcgtccgacg ggcggccagc tgagagcgat
      181 cggaggttac cccctcacaa cggccttggg cataggcagc gtagttggca tcggagttcg
      241 gaaatttaat ctcctgatat tggacaccca gctcgcgcat cgtgtccgcc agattgagtt
      301 ccgatgtggt gcccgtctcc acgcagattg acttgccctg aaaatcctgc agcgattgga
      361 tgccactatt gcgccgcacc atcaaccctt ggccgtcata gaaagtggtt ggcgcaaact
      421 caaggttgtt gccgcccttg gcatcgcgac tcagggtcca agtggtgttg cgcgagagca
      481 aatcgacttc accacttgct aaggctggaa accgttcaac agaatcgaga ctgcggtatt
      541 cgatcgcctt gggatcattg aacagtgctg cggcgatcgc cttacaaatg tcgacatcca
      601 agccactgta attgccctgg gaatcaagga agctaaagcc aggcagtcgg ccttcaactc
      661 cgcagagcaa cttaccgcgc gcctgcactt ggttgaggcg actattgctc tcagtctcat
      721 tgccacccaa gctacgacag gctgctagcg gtaaaaagca aagcagtacc aaaagaaagc
      781 gcgaggcgat cgcaggcatg ggtccgccgg ataaaaggat cgccttcatt caatcacgtc
      841 ttgccggttc taccagcctg cccagccaac ggctgagccg aatttgatct gcatgggatc
      901 ccaaacctga tctgagaaga gcattgcaaa gcactacggc tttctttcat tcctcttaag
      961 gccgatcgac cgccgtgatc ccctgtctat agtcagggca gtcacacggg ataataccgc
     1021 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     1081 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     1141 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     1201 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     1261 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     1321 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     1381 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     1441 ctttcgtctt cgaataaata cctgtgacgg aagatcactt cgcagaataa ataaatcctg
     1501 gtgtccctgt tgataccggg aagccctggg ccaacttttg gcgaaaatga gacgttgatc
     1561 ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg ggcgtatttt
     1621 ttgagttatc gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat
     1681 ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag
     1741 ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg
     1801 taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga
     1861 atgctcatcc ggaattccgt atggcaatga aagacggtga gctggtgata tgggatagtg
     1921 ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg
     1981 aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg
     2041 gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca
     2101 atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg
     2161 cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg
     2221 cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat
     2281 tacaacagta ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc agttattggt
     2341 gcccttaaac gcctggtgct acgcctgaat aagtgataat aagcggatga atggcagaaa
     2401 ttcgaaagca aattcgaccc ggtcgtcggt tcagggcagg gtcgttaaat agccgcttat
     2461 gtctattgct ggtttaccgg tttattgact accggaagca gtgtgaccgt gtgcttctca
     2521 aatgcctgag gccagtttgc tcaggctctc cccgtggagg taataattga cgatatgatc
     2581 gactgctagc tcgagcccgc gacatcttcc tgctccagaa gctcgaaatt cgagctcggt
     2641 acccggggat ctgggccgcg ggtcatggct gcgccccgac acccgccaac acccgctgac
     2701 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     2761 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagcagatc
     2821 aattcgcgcg cgaaggcgaa gcggcatgca tttacgttga caccatcgaa tggtgcaaaa
     2881 cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga
     2941 aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc
     3001 gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga
     3061 tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt
     3121 tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg
     3181 cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa
     3241 gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc
     3301 tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta
     3361 atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt attattttct
     3421 cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa
     3481 tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc
     3541 ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg
     3601 ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga
     3661 tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc
     3721 tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga taccgaagac agctcatgtt
     3781 atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg
     3841 accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct
     3901 cactggtgaa aagaaaaacc accctggcgc ccaatacgca aaccgcctct ccccgcgcgt
     3961 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     4021 cgcaacgcaa ttaatgtaag ttagcgcgaa ttgatctggt ttgacagctt atcatcgact
     4081 gcacggtgca ccaatgcttc tggcgtcagg cagccatcgg aagctgtggt atggctgtgc
     4141 aggtcgtaaa tcactgcata attcgtgtcg ctcaaggcgc actcccgttc tggataatgt
     4201 tttttgcgcc gacatcataa cggttctggc aaatattctg aaatgagctg ttgacaatta
     4261 atcatccggc tcgtataatg tgtggaattg tgagcggata acaatttcac acaggaaaca
     4321 gaccatggaa ttcatgtcca aaattcgtgt tggaattaac ggctttggcc gtattggtcg
     4381 tctctgctta cgggctggat ttgatgatcc gcaactcgat tttgttgcga ttaacgacct
     4441 cgttccggcc aaaaatcttg cctatcttct tcactacgac tccacccacg gccgcctaca
     4501 acaaacagtg cgggcagaag aagatcgcat ttgtgccggc gatcgctgca ttcaatgttt
     4561 ctccgaaact gatccggcag ctattccttg gggggagtta ggggttgatg tggtgattga
     4621 ggctacgggg cgcttcaccg attacgctgc cgccagtcgc cacctggaag caggcgctca
     4681 acgggtagtg atttcggccc cgacctcgga gaaagatccc gatcgcgttc ccaccttcgt
     4741 ctacggcgtc aatcatcaga actacgaccc gcaacagcat cgcattctct ccaacgccag
     4801 ctgcaccacc aactgcctgg ctccgatcgc caaggtgctt caagagcact gcgggattga
     4861 agacggtctg atgactacca tccatgcggt caccgccacc caaccaactg tcgatggacc
     4921 cagccgtaaa gatatgcgcg gcgggcgcag tgcggctcaa aacatcattc cggccagtac
     4981 gggcgctgcc aaagccgtgg gcttggtgct gcccgaactc cagggtcacc tgaccggtat
     5041 ggcgttgcgg gttccgacgg cggatgtctc ggtggtggat ctgaccttcc gtagccgcca
     5101 atccaccacc tatgccgcca tctgtgaggt gatgcaaaag gcggccaaca cccaactcca
     5161 aggcatcctg ggctataccg aggatgacct cgtttcgagt gactttattg gcgatgctcg
     5221 gtccagcatt tttgatgcca cggcaggcat ggctctcaac gatcgcttct tcaaagtgat
     5281 cgcttggtac gacaacgaat ggggatacag caaccgcatg ttggatctga
//
