LOCUS       sequence-80932-e        6295 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Replaces the psbA2 gene in Synechocystis with the Ptrc promoter - 
            PHLS gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6295)
  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 6295)
  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: psbA2-Ptrc-PHLS (d)
FEATURES             Location/Qualifiers
     source          1..6295
                     /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        join(1091..1096,1097..1113,1114..1120)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1129..1145
                     /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        2800..2902
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2903..3562
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2969..2988)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4110..4128)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4149..4165)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4149..4165)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4162..4184)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4173..4189
                     /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(4197..4203,4204..4221,4222..4227))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4242..4263
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4380..4397)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4551..5139)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4631..4650)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5310..6101,6102..6170))
                     /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     5933..5952
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6171..6275)
                     /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 gaggtggata tttgctgggg gttaatgaaa cattgtggcg gaacccaggg
      721 acaatgtgac caaaaaattc agggatatca ataagtatta ggtatatgga tcataattgt
      781 atgcccgact attgcttaaa ctgactgacc actgacctta agagtaatgg cgtgcaaggc
      841 ccagtgataa atttcattat ttttcattat ttcatctcca ttgtccctga aaatcagttg
      901 tgtcgcccct ctacacagcc cagaactatg gtaaaggcgc acgaaaaacc gccaggtaaa
      961 ctcttctcaa cccccaaaac gccctctgtt tacccatgga aaaaacgaca attacaagaa
     1021 agtaaaactt atgtcatcta taagcttcgt gtatattaac ttcctgttac aaagattctg
     1081 aaatgagctg ttgacaatta atcatccggc tcgtataatg tgtggaaatt gtgagcggat
     1141 aacaattagg aggtaaacat atgtgtagtt tgcaagtttc tgatcctatt cctaccggac
     1201 gccgttccgg tggttatccc ccggccttat gggatttcga tactattcaa tccctgaata
     1261 ccgaatataa gggcgaacgt cacatgcgtc gggaagaaga cttaattggt caagttcggg
     1321 aaatgttggt gcacgaagta gaagatccca ctccccagtt ggaattcatt gacgatctgc
     1381 ataaattggg catttcctgc cattttgaaa acgagattct gcaaattctc aaatccattt
     1441 atctcaacca aaactataaa cgggacctct attctaccag tttagccttc cgtctcttgc
     1501 gtcaatacgg gtttatcttg ccgcaggaag tttttgactg ctttaaaaac gaagaaggta
     1561 cggattttaa acccagcttc ggccgggata ttaagggtct gttacagttg tacgaagcct
     1621 cctttttgtc ccggaagggg gaagaaactt tacaactcgc ccgcgaattt gctaccaaaa
     1681 tcttgcaaaa ggaagtcgat gaacgggaat ttgctactaa aatggaattt cccagtcact
     1741 ggaccgtaca aatgcctaac gctcggcctt ttatcgatgc ctatcgtcgg cgtcccgaca
     1801 tgaaccccgt ggttctggaa ctcgccattc tcgataccaa tatcgtgcaa gctcagtttc
     1861 aagaagaatt gaaggagacc tcccgttggt gggaaagcac ggggattgtt caagaactgc
     1921 cgtttgttcg ggaccggatt gtggaaggtt atttttggac cattggtgtt actcaacgcc
     1981 gtgaacacgg ttacgaacgt attatgacgg ccaaagtcat cgctttggtg acctgtttgg
     2041 atgatattta tgacgtatat ggcactattg aagaattgca actcttcacc tctacgattc
     2101 agcgttggga tttggagtct atgaagcagt taccgactta tatgcaggta agcttcctgg
     2161 ccttgcacaa ttttgtaacc gaagtggcct atgatacgct gaagaaaaag ggctacaact
     2221 ctacccccta tttgcggaag acttgggtgg atttggtcga aagttacatt aaggaagcca
     2281 cttggtacta taatgggtac aaaccctcta tgcaggaata cctcaacaac gcctggatct
     2341 ctgtgggcag catggctatt ttgaatcatt tgttttttcg ctttactaat gaacgcatgc
     2401 ataagtaccg ggacatgaat cgtgtatcct ctaatattgt gcggttagcc gacgatatgg
     2461 gaacctcttt ggccgaagtt gaacgcggtg acgtgcccaa agctatccaa tgttacatga
     2521 atgaaacgaa cgcctctgag gaggaggccc gcgaatatgt gcggcgcgtt atccaggaag
     2581 aatgggaaaa actgaacact gaactgatgc gcgacgacga cgatgacgat gatttcacct
     2641 taagtaaata ctactgcgaa gtcgttgcta acctgacccg gatggctcag ttcatttacc
     2701 aagatggttc cgatgggttt gggatgaaag attccaaagt aaatcgttta ctgaaagaaa
     2761 cgctgattga gcgctatgag tgaagatctg cggccgcgtt gatcggcacg taagaggttc
     2821 caactttcac cataatgaaa taagatcact accgggcgta ttttttgagt tatcgagatt
     2881 ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca ccgttgatat
     2941 atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta
     3001 taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca
     3061 caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt
     3121 ccgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac
     3181 cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt
     3241 ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta
     3301 tttccctaaa gggtttattg agaatatgtt tttcgtctca gccaatccct gggtgagttt
     3361 caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat
     3421 gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca
     3481 tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga
     3541 tgagtggcag ggcggggcgt aattttttta aggcagttat tggtgccctt aaacgcctgg
     3601 ggatccttgg tgtaatgcca actgaataat ctgcaaattg cactctcctt caatgggggg
     3661 tgctttttgc ttgactgagt aatcttctga ttgctgatct tgattgccat cgatcgccgg
     3721 ggagtccggg gcagttacca ttagagagtc tagagaatta atccatcttc gatagaggaa
     3781 ttatggggga agaacctgtg ccggcggata aagcattagg caagaaattc aagaaaaaaa
     3841 atgcctcctg gagcattgaa gaaagcgaag ctctgtaccg ggttgaggcc tggggggcac
     3901 cttattttgc cattaatgcc gctggtaaca taaccgtctc tcccaacggc gatcggggcg
     3961 gttcgttaga tttgttggaa ctggtggaag ccctgcggca aagaaagctc ggcttacccc
     4021 tattaattcg tttttccgat attttggccg atcgcctaga gcgattgaat agttgttttg
     4081 ccaaggcgat cgagctccag cttttgttcc ctttagtgag ggttaatttc gagcttggcg
     4141 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4201 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4261 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4321 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4381 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4441 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4501 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4561 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4621 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4681 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4741 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4801 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4861 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4921 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4981 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5041 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5101 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5161 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5221 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5281 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5341 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5401 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5461 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5521 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5581 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5641 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5701 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5761 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5821 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5881 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5941 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6001 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6061 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6121 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6181 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6241 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgc
//
