LOCUS       sequence-137697-        7093 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Picea sitchensis PHLS 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 7093)
  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 7093)
  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: CpcB-Picea-sitchensisPHLS_CpcA
FEATURES             Location/Qualifiers
     source          1..7093
                     /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        3484..3586
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3587..4246
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3653..3672)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4908..4926)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4947..4963)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4947..4963)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4960..4982)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4971..4987
                     /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(4995..5001,5002..5019,5020..5025))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5040..5061
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5178..5195)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5349..5937)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5429..5448)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6108..6899,6900..6968))
                     /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     6731..6750
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6969..7073)
                     /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 gccgtagccc
     1741 atatgtccag ccctgtgtct gatgacggag ttcaaagacg gacgggtggc taccattcca
     1801 acttatggaa tgatgatatt attcaatttt taagtacgac ttatggtgaa cctgcctatc
     1861 gagaacgagg cgaacgcttg attgatgaag ttaaaaatat gtttaattcg atttctatgg
     1921 aagatgttga gttttcacca ctgaatgact tgatccagcg tctgtggatc gtcgattccg
     1981 tcgaacgttt ggggatcgat cgccatttta aaaacgaaat taagtcaacc ctcgactatg
     2041 tttactccta ctggacgcag aagggcatag ggtgtggcat cgaaagcgta gtaccagact
     2101 taaacagtac ggcactggga ttgcgcacgc tccgtcttca tggatatcct gtaagcgctg
     2161 aagtccttaa acacttccaa aatcaaaatg gtcagtttgc ctgtagcccg tcagagacgg
     2221 aaggcgaaat gcggtccatt gtaaacttat atcgggcttc cttaattgca tttcccggcg
     2281 aaaaagtgat ggaagaagcc gaaatttttt ccacgaaata cttgaaggaa gcactgcaga
     2341 aaattcccgt aagtagcctg agccgtgaaa tcggggatgt attggaacaa gactggcata
     2401 ctaacttgcc acgattagaa gctcgaaatt acattgatgt cttcggtcaa gatactaaag
     2461 atacgaaact gtacatgaaa accgagaaac ttctggaatt agctaaactt gaattcaaca
     2521 tttttcagtc actccaaaaa acggaactgg actccttgtt gcgctggtgg aaagatagtg
     2581 gctttcatca tatcaccttt tcccgacatc tacatgtgga atattacaca ttggcctcgt
     2641 gtatcgccat tgagccccag cactctagat ttcgccttgg ctttgccaaa gcgtgtcatg
     2701 tgataaccat cctggatgac atgtacgatg tttttggcac catcgatgag ttagagcttt
     2761 ttaccgcaca gattaagcgc tgggacccgt cagctaccga ttgtttgccc aaatatatga
     2821 aacgcatgta tatgatctta tacgatatgg tgaacgaaat gtctcgtgaa gccgaaaccg
     2881 cccagggaag ggataccctt aattacgcca gacaagcctg ggaagatttt atcgattcct
     2941 atatgcagga agcaaagtgg atcgctaccg ggtaccttcc cacgttcgat gagtacttcg
     3001 agaacggtaa ggtttcctcc ggccatcgtg ttgccgcgtt gcagcccatc cttacaatgg
     3061 atatcccctt tccccatgac atattgaaag aagttgattt tccgtccaaa ctcaatgatc
     3121 tagcctctgc catcttgcgt ctacgcggcg atacgaggtg ttataaagcc gaccgtgctc
     3181 gtggtgaaga ggcctcctgc atttcctgtt atatgaaaga caacccgggc gctactgagg
     3241 aggacgcttt gagccatatt aacgcggtga tatccgatgt cattaaagga ttaaactggg
     3301 aactactcaa tcccaatagt tccgtgccga tttcgtcaaa aaagcacgtt tttgacgtgt
     3361 cgcgcgctct gcactatgga tataagtatc gtgatgggta tagcgtgtca aatattgaaa
     3421 caaagtcatt ggtcatgaga actctcctag aatcagtacc attttagaga tctgcggccg
     3481 cgttgatcgg cacgtaagag gttccaactt tcaccataat gaaataagat cactaccggg
     3541 cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat
     3601 cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt
     3661 tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt
     3721 aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg
     3781 cctgatgaat gctcatccgg aattccgtat ggcaatgaaa gacggtgagc tggtgatatg
     3841 ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct
     3901 ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc
     3961 gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt
     4021 ctcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa
     4081 cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat
     4141 gccgctggcg attcaggttc atcatgccgt ctgtgatggc ttccatgtcg gcagaatgct
     4201 taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag
     4261 ttattggtgc ccttaaacgc ctggggatcc tctggttatt ttaaaaacca actttactca
     4321 ggttccatac ccgagaaaat ccagcttaaa gctgacatat ctaggaaaat tttcacattc
     4381 taacgggaga taccagaaca atgaaaaccc ctttaactga agccgtttcc accgctgact
     4441 ctcaaggtcg ctttctgagc agcaccgaat tgcaaattgc tttcggtcgt ctacgtcaag
     4501 ctaatgctgg tttgcaagcc gctaaagctc tgaccgacaa tgcccagagc ttggtaaatg
     4561 gtgctgccca agccgtttat aacaaattcc cctacaccac ccaaacccaa ggcaacaact
     4621 ttgctgcgga tcaacggggt aaagacaagt gtgcccggga catcggctac tacctccgca
     4681 tcgttaccta ctgcttagtt gctggtggta ccggtccttt ggatgagtac ttgatcgccg
     4741 gtattgatga aatcaaccgc acctttgacc tctcccccag ctggtatgtt gaagctctga
     4801 aatacatcaa agctaaccac ggcttgagtg gcgatgcccg tgacgaagct aattcctacc
     4861 tcgattacgc catcaatgct ctgagctagg agctccagct tttgttccct ttagtgaggg
     4921 ttaatttcga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     4981 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     5041 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5101 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5161 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5221 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5281 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5341 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5401 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5461 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5521 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5581 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5641 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5701 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5761 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     5821 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5881 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5941 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6001 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6061 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6121 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6181 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6241 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6301 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6361 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6421 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6481 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6541 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6601 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6661 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6721 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6781 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6841 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6901 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6961 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7021 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7081 ccccgaaaag tgc
//
