LOCUS       sequence-137695-        7963 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Nicotiana attenuata GLS as a fusion with the CpcB 
            protein plus the CmR cassete in Synechocystis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7963)
  AUTHORS   Formighieri C, Melis A
  TITLE     Cyanobacterial production of plant essential oils.
  JOURNAL   Planta. 2018 Oct;248(4):933-946. doi: 10.1007/s00425-018-2948-0. 
            Epub 2018 Jul 4.
  PUBMED    29974209
REFERENCE   2  (bases 1 to 7963)
  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"; doi: 
            "10.1007/s00425-018-2948-0"; journalName: "Planta"; date: "2018-10";
            volume: "248"; issue: "4"; pages: "933-946"
COMMENT     Sequence Label: CpcBfull-NaGLS fusion_CpcA
FEATURES             Location/Qualifiers
     source          1..7963
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(33..461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(143..164)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     355..374
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     588..610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     602..619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        4354..4456
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             4457..5116
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4523..4542)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(5778..5796)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5817..5833)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5817..5833)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5830..5852)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5841..5857
                     /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(5865..5871,5872..5889,5890..5895))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5910..5931
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6048..6065)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6219..6807)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6299..6318)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6978..7769,7770..7838))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7601..7620
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7839..7943)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
       61 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
      121 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
      181 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
      241 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
      301 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
      361 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
      421 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tgggtaccgg
      661 gccccccctc gagaagagtc cctgaatatc aaaatggtgg gataaaaagc tcaaaaagga
      721 aagtaggctg tggttcccta ggcaacagtc ttccctaccc cactggaaac taaaaaaacg
      781 agaaaagttc gcaccgaaca tcaattgcat aattttagcc ctaaaacata agctgaacga
      841 aactggttgt cttcccttcc caatccagga caatctgaga atcccctgca acattactta
      901 acaaaaaagc aggaataaaa ttaacaagat gtaacagaca taagtcccat caccgttgta
      961 taaagttaac tgtgggattg caaaagcatt caagcctagg cgctgagctg tttgagcatc
     1021 ccggtggccc ttgtcgctgc ctccgtgttt ctccctggat ttatttaggt aatatctctc
     1081 ataaatcccc gggtagttaa cgaaagttaa tggagatcag taacaataac tctagggtca
     1141 ttactttgga ctccctcagt ttatccgggg gaattgtgtt taagaaaatc ccaactcata
     1201 aagtcaagta ggagattaat tcaatgttcg acgtattcac tcgggttgtt tcccaagctg
     1261 atgctcgcgg cgagtacctc tctggttctc agttagatgc tttgagcgct accgttgctg
     1321 aaggcaacaa acggattgat tctgttaacc gcatcaccgg taatgcttcc gctatcgttt
     1381 ccaacgctgc tcgtgctttg ttcgccgaac agccccaatt aatccaaccc ggtggaaacg
     1441 cctacaccag ccgtcgtatg gctgcttgtt tgcgtgacat ggaaatcatc ctccgctatg
     1501 ttacctacgc aaccttcacc ggcgacgctt ccgttctaga agatcgttgc ttgaacggtc
     1561 tccgtgaaac ctacgttgcc ctgggtgttc ccggtgcttc cgtagctgct ggcgttcaaa
     1621 aaatgaaaga agctgccctg gacatcgtta acgatcccaa tggcatcacc cgtggtgatt
     1681 gcagtgctat cgttgctgaa atcgctggtt acttcgaccg cgccgctgct gccgtagcca
     1741 ctagtgattc ctcctccatt gtgtcttcca ttgaatctct agtcaatgag atcaagaaag
     1801 atatgttctc aaaccaagaa tttaacactt ttgttacccc aatatctgcc tatgacactg
     1861 cttggttggc catgatttct tataataatc aagaaaaagc cactaatgat cattcttttt
     1921 ctggccctat gtttgagagt tgtttaaatt ggattctcaa caaccaaaat gagcaaggat
     1981 tttggggaga atccaatggt gaaaatcttc ctaccattta tgctctccct actactcttg
     2041 cttgtatgat agcactcaag aaatgggatg tgggtgaaac caacatcaaa aaaggtttga
     2101 aatttattca tgctaatacg gaaatacttc tgaaagagaa ttctcaacaa tttccacgtt
     2161 ggtttaccat tgttttccct gcaatggttc aacttgctaa atcagtaggc ctggagatta
     2221 tttttgtgaa tggatcagaa agattacttt ctgacgtggt catgaagagg agagtaattc
     2281 ttgaaagtga aaagctaatg gatgtgacag cagggcatga tcaacctctt ttggcatatc
     2341 tagagagctt gccaaattat tttgtggatc aaaaagacca aatcctaaaa catttaagtg
     2401 aagatggttc tttgtttcaa tctccctctg ctacagcaca agcattcatg tccacaggga
     2461 accaaaaatg tcttgaatat ctcatgtcca ttgttcacaa gtgtccaaat ggagtacctt
     2521 caagatttcc agtggatgaa gagctaataa atctttgcat gattgaccac attcagagaa
     2581 tgggattggc taaccacttt aacaaagaga ttgaacagat tcttggccaa gtttataaga
     2641 atcaaatgaa taaccagaat gagtacttat cagaaataag tactttacct gagagattat
     2701 acaaggactc tttggctttt cgacttctcc gtttgcaagg ctataaaata aatcaaggaa
     2761 gtttttgttg gtttgttcat caaccgcaga taagggccta catagaaaag aatcaagaac
     2821 gttttacata tgtgatgtac aatgtgtaca gagctacaga tcttatgttc aatggagaat
     2881 ctgaaatgga gaaagtacga tcttttgcca gaaattttct tgaaagttca atggaactcg
     2941 tgagcaatga ggacaactta ttactacttc cttcacttca aaaagtgatt gagcatgagt
     3001 tgaatgttcc atgggttgct cgattagaac atcttgatca cagattatgg attgaattaa
     3061 gccaatctct ccctctttca attggaaaat ctgcagatta ttggttgtct tgtctgcaca
     3121 atgacaaatt ggtgaaacta gcagtgcaaa attacgaata tcgacagtca gtttacagga
     3181 cagaattgga agaactcaag aggtggtcga aagagaaaga cctcgtggac attggatttg
     3241 ggcgagagaa aactacatat tcctattttg caagtgcagc tagcagcagc agttttctac
     3301 cttttgattc ttttcttcga ttggttgttg caaaatgttc catagttatt acagttgctg
     3361 atgattttta tgatgaggaa gcctctttaa atgacttgaa aatcttaacc gacgctgtac
     3421 agaggtggga tggaagtaac cttgatggcc ctagtaagat catatttgat gcactcgatg
     3481 atcttgtgtg cgatgttgct aagttgtacc accttcgaca cgcaatcgac atcacttcag
     3541 aacttcgacg tcagtggcag gagacatttc ttgcatggat gatggaaagc acgtggagta
     3601 acaatgtagc catgccatct aggaatgaat acctagaaat tggcatgata tcaattggtg
     3661 cacatatttt ggttcttcat gctgcttctc ttgaaaatcc aagtttgcca agggaaaaac
     3721 ttaagccaat tgatggccaa tatgaaaata ttacaaagtt gcttatggtc accactcgtt
     3781 tgttgaatga tatccagagc tatcaaaaag aatgtgaagt agggaaaatg aactacacat
     3841 tacttcacat gaatgaaaat ccaggggcac aacttgatga ttcaattgaa tatgtgaaag
     3901 agattttggc caacaagaag aaggaatttc ttgaaaatgt tctaatggat ggattcaatg
     3961 atatgccaaa gacatgcaaa ctacttcatc tttcttgcct aaatgtgttt cacatgttct
     4021 tcaattcttg caatttattt gataccaaag cagcaattct tgaagatata atgagagcta
     4081 tttacattcc tcttcaagaa caaacctcaa ttccatcatt aaaaacttct ccaattttaa
     4141 cacctcaaga aaagaagaag aagaaggaga agaagattga gaatattcca actaaagttt
     4201 ctgcttgtct caatgggaaa aacttcaaac atcaagttgg cattggaata agttttgttg
     4261 gaagtaaagc tcctaagaat catccttttg gttggaatac aaaaggattt gcttcacata
     4321 agctaagttc atgtttcatg taagcggccg cgttgatcgg cacgtaagag gttccaactt
     4381 tcaccataat gaaataagat cactaccggg cgtatttttt gagttatcga gattttcagg
     4441 agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca
     4501 atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca
     4561 gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt
     4621 ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg aattccgtat
     4681 ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt
     4741 ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca
     4801 gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc
     4861 taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag
     4921 ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa
     4981 atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt
     5041 ctgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg
     5101 gcagggcggg gcgtaatttt tttaaggcag ttattggtgc ccttaaacgc ctggggatcc
     5161 tctggttatt ttaaaaacca actttactca ggttccatac ccgagaaaat ccagcttaaa
     5221 gctgacatat ctaggaaaat tttcacattc taacgggaga taccagaaca atgaaaaccc
     5281 ctttaactga agccgtttcc accgctgact ctcaaggtcg ctttctgagc agcaccgaat
     5341 tgcaaattgc tttcggtcgt ctacgtcaag ctaatgctgg tttgcaagcc gctaaagctc
     5401 tgaccgacaa tgcccagagc ttggtaaatg gtgctgccca agccgtttat aacaaattcc
     5461 cctacaccac ccaaacccaa ggcaacaact ttgctgcgga tcaacggggt aaagacaagt
     5521 gtgcccggga catcggctac tacctccgca tcgttaccta ctgcttagtt gctggtggta
     5581 ccggtccttt ggatgagtac ttgatcgccg gtattgatga aatcaaccgc acctttgacc
     5641 tctcccccag ctggtatgtt gaagctctga aatacatcaa agctaaccac ggcttgagtg
     5701 gcgatgcccg tgacgaagct aattcctacc tcgattacgc catcaatgct ctgagctagg
     5761 agctccagct tttgttccct ttagtgaggg ttaatttcga gcttggcgta atcatggtca
     5821 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     5881 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5941 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     6001 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     6061 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     6121 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6181 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6241 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6301 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6361 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6421 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6481 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6541 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6601 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     6661 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6721 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6781 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6841 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6901 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6961 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     7021 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     7081 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     7141 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     7201 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     7261 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     7321 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     7381 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     7441 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7501 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7561 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7621 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7681 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7741 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7801 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7861 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7921 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgc
//
