LOCUS       sequence-80936-e        6429 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Replaces the cpc operon in Synechocystis with the PHLS gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6429)
  AUTHORS   Formighieri C, Melis A
  TITLE     Regulation of beta-phellandrene synthase gene expression, 
            recombinant protein accumulation, and monoterpene hydrocarbons 
            production in Synechocystis transformants.
  JOURNAL   Planta. 2014 Aug;240(2):309-24. doi: 10.1007/s00425-014-2080-8. Epub
            2014 May 20.
  PUBMED    24838596
REFERENCE   2  (bases 1 to 6429)
  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/s00425-014-2080-8"; journalName: "Planta"; date: "2014-08";
            volume: "240"; issue: "2"; pages: "309-24"
COMMENT     Sequence Label: cpc-PHLS (g)
FEATURES             Location/Qualifiers
     source          1..6429
                     /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        2869..2971
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2972..3631
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3038..3057)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     regulatory      3995..4004
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4244..4262)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4283..4299)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4283..4299)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4296..4318)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4307..4323
                     /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(4331..4337,4338..4355,4356..4361))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4376..4397
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4514..4531)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4685..5273)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4765..4784)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5444..6235,6236..6304))
                     /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     6067..6086
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6305..6409)
                     /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 tcaatgcata tgtgtagttt gcaagtttct gatcctattc
     1261 ctaccggacg ccgttccggt ggttatcccc cggccttatg ggatttcgat actattcaat
     1321 ccctgaatac cgaatataag ggcgaacgtc acatgcgtcg ggaagaagac ttaattggtc
     1381 aagttcggga aatgttggtg cacgaagtag aagatcccac tccccagttg gaattcattg
     1441 acgatctgca taaattgggc atttcctgcc attttgaaaa cgagattctg caaattctca
     1501 aatccattta tctcaaccaa aactataaac gggacctcta ttctaccagt ttagccttcc
     1561 gtctcttgcg tcaatacggg tttatcttgc cgcaggaagt ttttgactgc tttaaaaacg
     1621 aagaaggtac ggattttaaa cccagcttcg gccgggatat taagggtctg ttacagttgt
     1681 acgaagcctc ctttttgtcc cggaaggggg aagaaacttt acaactcgcc cgcgaatttg
     1741 ctaccaaaat cttgcaaaag gaagtcgatg aacgggaatt tgctactaaa atggaatttc
     1801 ccagtcactg gaccgtacaa atgcctaacg ctcggccttt tatcgatgcc tatcgtcggc
     1861 gtcccgacat gaaccccgtg gttctggaac tcgccattct cgataccaat atcgtgcaag
     1921 ctcagtttca agaagaattg aaggagacct cccgttggtg ggaaagcacg gggattgttc
     1981 aagaactgcc gtttgttcgg gaccggattg tggaaggtta tttttggacc attggtgtta
     2041 ctcaacgccg tgaacacggt tacgaacgta ttatgacggc caaagtcatc gctttggtga
     2101 cctgtttgga tgatatttat gacgtatatg gcactattga agaattgcaa ctcttcacct
     2161 ctacgattca gcgttgggat ttggagtcta tgaagcagtt accgacttat atgcaggtaa
     2221 gcttcctggc cttgcacaat tttgtaaccg aagtggccta tgatacgctg aagaaaaagg
     2281 gctacaactc taccccctat ttgcggaaga cttgggtgga tttggtcgaa agttacatta
     2341 aggaagccac ttggtactat aatgggtaca aaccctctat gcaggaatac ctcaacaacg
     2401 cctggatctc tgtgggcagc atggctattt tgaatcattt gttttttcgc tttactaatg
     2461 aacgcatgca taagtaccgg gacatgaatc gtgtatcctc taatattgtg cggttagccg
     2521 acgatatggg aacctctttg gccgaagttg aacgcggtga cgtgcccaaa gctatccaat
     2581 gttacatgaa tgaaacgaac gcctctgagg aggaggcccg cgaatatgtg cggcgcgtta
     2641 tccaggaaga atgggaaaaa ctgaacactg aactgatgcg cgacgacgac gatgacgatg
     2701 atttcacctt aagtaaatac tactgcgaag tcgttgctaa cctgacccgg atggctcagt
     2761 tcatttacca agatggttcc gatgggtttg ggatgaaaga ttccaaagta aatcgtttac
     2821 tgaaagaaac gctgattgag cgctatgagt gaagatctgc ggccgcgttg atcggcacgt
     2881 aagaggttcc aactttcacc ataatgaaat aagatcacta ccgggcgtat tttttgagtt
     2941 atcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg gatataccac
     3001 cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca
     3061 atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa
     3121 aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga tgaatgctca
     3181 tccggaattc cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc
     3241 ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca
     3301 cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa
     3361 cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg
     3421 ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt
     3481 tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca
     3541 ggttcatcat gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca
     3601 gtactgcgat gagtggcagg gcggggcgta atttttttaa ggcagttatt ggtgccctta
     3661 aacgcctggg gatccgctat tttgttaatt actatttgag ctgagtgtaa aataccttac
     3721 ttactcaaaa gcattaacta accataacaa tgactaatct ctttttttga ttgaactcca
     3781 aactagaata gccatcgagt cagtccattt agttcattat tagtgaaagt ttgttggcgg
     3841 tgggttatcc gttgataaac caccgttttt gtttgggcaa agtaacgatt tgatgcagtg
     3901 atgggtttaa agataatccc gtttgaggaa atcctgcagg acgacgggaa ctttaacctg
     3961 accgctgctg ggttcgtaat aattttctaa aattgccgcc atggtgcgcc cgatcgccaa
     4021 accggaaccg ttgagagtgt gaacaaattg ggtgcctttt ttgccctttt ccttgtagcg
     4081 aatgttggcc cgacgggctt ggaaatcgtg gaagttagaa caactggaaa tttcccggta
     4141 ggtgttagcc gatggtaacc aaacttccaa gtcgtagcat ttagccgctc caaaacctaa
     4201 atcaccggta cataattcca ccactgagct ccagcttttg ttccctttag tgagggttaa
     4261 tttcgagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     4321 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     4381 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     4441 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4501 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4561 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4621 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4681 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4741 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4801 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4861 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4921 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4981 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5041 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5101 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     5161 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5221 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5281 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5341 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5401 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5461 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5521 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5581 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5641 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5701 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5761 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5821 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5881 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5941 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6001 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6061 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6121 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6181 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6241 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6301 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6361 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6421 gaaaagtgc
//
