LOCUS       sequence-80935-e        6324 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Replaces the psbA2 gene in Synechocystis with the Ptrc 
            promoter-T7-g10-PHLS gene .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6324)
  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 6324)
  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-T7-g10-PHLS (e)
FEATURES             Location/Qualifiers
     source          1..6324
                     /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        2829..2931
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2932..3591
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2998..3017)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4139..4157)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4178..4194)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4178..4194)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    4202..4218
                     /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(4226..4232,4233..4250,4251..4256))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4271..4292
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4409..4426)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4580..5168)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4660..4679)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5339..6130,6131..6199))
                     /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     5962..5981
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6200..6304)
                     /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 ccagtgatca 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 aacaatttca tgttaacttt aagaaggaga tatagttatg gctagccata tgtgtagttt
     1201 gcaagtttct gatcctattc ctaccggacg ccgttccggt ggttatcccc cggccttatg
     1261 ggatttcgat actattcaat ccctgaatac cgaatataag ggcgaacgtc acatgcgtcg
     1321 ggaagaagac ttaattggtc aagttcggga aatgttggtg cacgaagtag aagatcccac
     1381 tccccagttg gaattcattg acgatctgca taaattgggc atttcctgcc attttgaaaa
     1441 cgagattctg caaattctca aatccattta tctcaaccaa aactataaac gggacctcta
     1501 ttctaccagt ttagccttcc gtctcttgcg tcaatacggg tttatcttgc cgcaggaagt
     1561 ttttgactgc tttaaaaacg aagaaggtac ggattttaaa cccagcttcg gccgggatat
     1621 taagggtctg ttacagttgt acgaagcctc ctttttgtcc cggaaggggg aagaaacttt
     1681 acaactcgcc cgcgaatttg ctaccaaaat cttgcaaaag gaagtcgatg aacgggaatt
     1741 tgctactaaa atggaatttc ccagtcactg gaccgtacaa atgcctaacg ctcggccttt
     1801 tatcgatgcc tatcgtcggc gtcccgacat gaaccccgtg gttctggaac tcgccattct
     1861 cgataccaat atcgtgcaag ctcagtttca agaagaattg aaggagacct cccgttggtg
     1921 ggaaagcacg gggattgttc aagaactgcc gtttgttcgg gaccggattg tggaaggtta
     1981 tttttggacc attggtgtta ctcaacgccg tgaacacggt tacgaacgta ttatgacggc
     2041 caaagtcatc gctttggtga cctgtttgga tgatatttat gacgtatatg gcactattga
     2101 agaattgcaa ctcttcacct ctacgattca gcgttgggat ttggagtcta tgaagcagtt
     2161 accgacttat atgcaggtaa gcttcctggc cttgcacaat tttgtaaccg aagtggccta
     2221 tgatacgctg aagaaaaagg gctacaactc taccccctat ttgcggaaga cttgggtgga
     2281 tttggtcgaa agttacatta aggaagccac ttggtactat aatgggtaca aaccctctat
     2341 gcaggaatac ctcaacaacg cctggatctc tgtgggcagc atggctattt tgaatcattt
     2401 gttttttcgc tttactaatg aacgcatgca taagtaccgg gacatgaatc gtgtatcctc
     2461 taatattgtg cggttagccg acgatatggg aacctctttg gccgaagttg aacgcggtga
     2521 cgtgcccaaa gctatccaat gttacatgaa tgaaacgaac gcctctgagg aggaggcccg
     2581 cgaatatgtg cggcgcgtta tccaggaaga atgggaaaaa ctgaacactg aactgatgcg
     2641 cgacgacgac gatgacgatg atttcacctt aagtaaatac tactgcgaag tcgttgctaa
     2701 cctgacccgg atggctcagt tcatttacca agatggttcc gatgggtttg ggatgaaaga
     2761 ttccaaagta aatcgtttac tgaaagaaac gctgattgag cgctatgagt gaagatctgc
     2821 ggccgcgttg atcggcacgt aagaggttcc aactttcacc ataatgaaat aagatcacta
     2881 ccgggcgtat tttttgagtt atcgagattt tcaggagcta aggaagctaa aatggagaaa
     2941 aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag
     3001 gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc
     3061 tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt
     3121 gcccgcctga tgaatgctca tccggaattc cgtatggcaa tgaaagacgg tgagctggtg
     3181 atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca
     3241 tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat
     3301 gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt
     3361 ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg
     3421 gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg
     3481 ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga
     3541 atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta atttttttaa
     3601 ggcagttatt ggtgccctta aacgcctggg gatccttggt gtaatgccaa ctgaataatc
     3661 tgcaaattgc actctccttc aatggggggt gctttttgct tgactgagta atcttctgat
     3721 tgctgatctt gattgccatc gatcgccggg gagtccgggg cagttaccat tagagagtct
     3781 agagaattaa tccatcttcg atagaggaat tatgggggaa gaacctgtgc cggcggataa
     3841 agcattaggc aagaaattca agaaaaaaaa tgcctcctgg agcattgaag aaagcgaagc
     3901 tctgtaccgg gttgaggcct ggggggcacc ttattttgcc attaatgccg ctggtaacat
     3961 aaccgtctct cccaacggcg atcggggcgg ttcgttagat ttgttggaac tggtggaagc
     4021 cctgcggcaa agaaagctcg gcttacccct attaattcgt ttttccgata ttttggccga
     4081 tcgcctagag cgattgaata gttgttttgc caaggcgatc gagctccagc ttttgttccc
     4141 tttagtgagg gttaatttcg agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     4201 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     4261 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     4321 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     4381 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     4441 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4501 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4561 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4621 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4681 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4741 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4801 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4861 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4921 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4981 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     5041 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5101 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5161 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5221 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     5281 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     5341 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     5401 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     5461 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5521 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5581 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5641 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5701 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5761 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5821 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5881 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5941 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6001 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6061 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6121 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6181 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     6241 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     6301 cgcgcacatt tccccgaaaa gtgc
//
