LOCUS       sequence-137699-        7090 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Pinus banksiana 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 7090)
  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 7090)
  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-Pinus-banksianaPHLS_CpcA
FEATURES             Location/Qualifiers
     source          1..7090
                     /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"
     CDS             complement(3342..3353)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        3481..3583
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3584..4243
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3650..3669)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4905..4923)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4944..4960)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4944..4960)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4957..4979)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4968..4984
                     /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(4992..4998,4999..5016,5017..5022))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5037..5058
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5175..5192)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5346..5934)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5426..5445)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6105..6896,6897..6965))
                     /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     6728..6747
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6966..7070)
                     /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 atatggtcag tagcacagcc agtgtttcaa atgatgatgg agtgcggaga cgcgtgggcg
     1801 attatcgata caatcactgg gatgaagacc tgatcgattc cttggccacc tcatacgaag
     1861 ctccttccta tttgaaacgt gcggatacgc tcgtcgaagc gattaaagat cggtttaata
     1921 gtatgggtgt agatgatggg gaaagaatgt ctccattaac tgatctctat caacgactgt
     1981 ggatggtaga tagtgtcgaa cgtttgggta tagatcgcca tttccaaaat gagattaaaa
     2041 gcgccttgga ctatgtgttt tcttattgga aagagaaagg catcggtcgt ggccgccaaa
     2101 gcgctgttac cgatctgaat tccaccgccc tgggcttacg tactttacga ctgcacgggt
     2161 accccgtgtc atccgatgtg ttggaaaatt ttaaagatca taacggtcag tttacgtgta
     2221 gcggcattca aacagaaggt gagattagag gcgtgttaaa cttgtttcgt gctagtctta
     2281 tcgccttccc cggcgaaaaa gtcatggaag aagccgaaat tttttccact atgtatctta
     2341 aacatgcctt gcaaaaaatt gcggtgtcca gtctgtctca ggaaatcgaa tacttactgg
     2401 agtatggctg gcataccaac ccgcctcgcc tggaagcgcg catgtatatg gaagtatttc
     2461 cccaagacac catctacgaa cagaaattag tggaactggc taaagtggag ttcaatatct
     2521 ttcattctct acagaagcgt gaactgcagt ctttgacccg atggtggaaa cattatgggt
     2581 tcccccaatt aagttttact agacatatac atgtagagta ttacacattt ggtagctgta
     2641 tagccactga tccgaaacaa tcggcatttc ggctctgttt tgccaaaatg tcatatttcg
     2701 taaccgtgct agatgatatc tacgacacgt atggaaccat ggaggaactg gaattgttta
     2761 ctgccgccat taaacgctgg gacccctccg ttgtcgattg tttaccggag tatatgaaag
     2821 gtgtatatat ggccgtttac gataccgtaa acgaaatggc gaaggaggcg gaaaaagtac
     2881 aaggtcgtga tacgctaaac tacgtgcgcc aggcctggga actctatatc gacgcttata
     2941 tgccggaagc caaatggatt tcaagtggct acctgcccac ttttcaagaa tacctcgata
     3001 attcgaaaat ctcttttggc actcggatca ccattcttca accaatttta acgttgggtg
     3061 agcccctgcc gcacgaaatt ctacaggaga ttgactttcc tgccaagttt aatgatctaa
     3121 tttccgtgat tctacggtta aagggcgaca ccaggtgtta caaagccgat cgagcgcgag
     3181 gcgaggaggc atcgtccgtc tcctgctata tgaaagataa cgcggggata accgaagaag
     3241 atgccattca ctgcattaat gatatggtta ataatctgct gaaagagttg aactgggaac
     3301 tgctgaaacc cgacagtaac gtgcccatca gttgccggaa agcggccttc gatatttgta
     3361 ggatttttca tcacggatac aaatatcggg atggctatgg cgacgcaact attgaagtta
     3421 aaaatttggt caaacgcacc gtattagaac ccgttcccct gtagagatct gcggccgcgt
     3481 tgatcggcac gtaagaggtt ccaactttca ccataatgaa ataagatcac taccgggcgt
     3541 attttttgag ttatcgagat tttcaggagc taaggaagct aaaatggaga aaaaaatcac
     3601 tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg aggcatttca
     3661 gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg cctttttaaa
     3721 gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc ttgcccgcct
     3781 gatgaatgct catccggaat tccgtatggc aatgaaagac ggtgagctgg tgatatggga
     3841 tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt catcgctctg
     3901 gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag atgtggcgtg
     3961 ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt ttttcgtctc
     4021 agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata tggacaactt
     4081 cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc
     4141 gctggcgatt caggttcatc atgccgtctg tgatggcttc catgtcggca gaatgcttaa
     4201 tgaattacaa cagtactgcg atgagtggca gggcggggcg taattttttt aaggcagtta
     4261 ttggtgccct taaacgcctg gggatcctct ggttatttta aaaaccaact ttactcaggt
     4321 tccatacccg agaaaatcca gcttaaagct gacatatcta ggaaaatttt cacattctaa
     4381 cgggagatac cagaacaatg aaaacccctt taactgaagc cgtttccacc gctgactctc
     4441 aaggtcgctt tctgagcagc accgaattgc aaattgcttt cggtcgtcta cgtcaagcta
     4501 atgctggttt gcaagccgct aaagctctga ccgacaatgc ccagagcttg gtaaatggtg
     4561 ctgcccaagc cgtttataac aaattcccct acaccaccca aacccaaggc aacaactttg
     4621 ctgcggatca acggggtaaa gacaagtgtg cccgggacat cggctactac ctccgcatcg
     4681 ttacctactg cttagttgct ggtggtaccg gtcctttgga tgagtacttg atcgccggta
     4741 ttgatgaaat caaccgcacc tttgacctct cccccagctg gtatgttgaa gctctgaaat
     4801 acatcaaagc taaccacggc ttgagtggcg atgcccgtga cgaagctaat tcctacctcg
     4861 attacgccat caatgctctg agctaggagc tccagctttt gttcccttta gtgagggtta
     4921 atttcgagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     4981 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     5041 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5101 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5161 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5221 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5281 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5341 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5401 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5461 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5521 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5581 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5641 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5701 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5761 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     5821 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5881 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5941 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6001 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6061 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6121 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6181 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6241 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     6301 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     6361 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6421 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6481 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6541 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6601 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6661 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6721 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6781 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6841 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6901 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6961 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7021 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7081 cgaaaagtgc
//
