LOCUS       sequence-80929-e        6337 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Replaces the psbA2 gene in Synechocystis with the PHLS gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6337)
  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 6337)
  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-PHLS (b)
FEATURES             Location/Qualifiers
     source          1..6337
                     /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"
     primer_bind     669..685
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     670..686
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2842..2944
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2945..3604
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3011..3030)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4152..4170)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4191..4207)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4191..4207)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4204..4226)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4215..4231
                     /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(4239..4245,4246..4263,4264..4269))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4284..4305
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4422..4439)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4593..5181)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4673..4692)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5352..6143,6144..6212))
                     /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     5975..5994
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6213..6317)
                     /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 gaggtcgacg gtatcgataa gcttgatatc gaattcagcg ttccagtgga
      721 tatttgctgg gggttaatga aacattgtgg cggaacccag ggacaatgtg accaaaaaat
      781 tcagggatat caataagtat taggtatatg gatcataatt gtatgcccga ctattgctta
      841 aactgactga ccactgacct taagagtaat ggcgtgcaag gcccagtgat caatttcatt
      901 atttttcatt atttcatctc cattgtccct gaaaatcagt tgtgtcgccc ctctacacag
      961 cccagaacta tggtaaaggc gcacgaaaaa ccgccaggta aactcttctc aacccccaaa
     1021 acgccctctg tttacccatg gaaaaaacga caattacaag aaagtaaaac ttatgtcatc
     1081 tataagcttc gtgtatatta acttcctgtt acaaagcttt acaaaactct cattaatcct
     1141 ttagactaag tttagtcagt tccaatctga acatcgacaa atacataagg aattataacc
     1201 atatgtgtag tttgcaagtt tctgatccta ttcctaccgg acgccgttcc ggtggttatc
     1261 ccccggcctt atgggatttc gatactattc aatccctgaa taccgaatat aagggcgaac
     1321 gtcacatgcg tcgggaagaa gacttaattg gtcaagttcg ggaaatgttg gtgcacgaag
     1381 tagaagatcc cactccccag ttggaattca ttgacgatct gcataaattg ggcatttcct
     1441 gccattttga aaacgagatt ctgcaaattc tcaaatccat ttatctcaac caaaactata
     1501 aacgggacct ctattctacc agtttagcct tccgtctctt gcgtcaatac gggtttatct
     1561 tgccgcagga agtttttgac tgctttaaaa acgaagaagg tacggatttt aaacccagct
     1621 tcggccggga tattaagggt ctgttacagt tgtacgaagc ctcctttttg tcccggaagg
     1681 gggaagaaac tttacaactc gcccgcgaat ttgctaccaa aatcttgcaa aaggaagtcg
     1741 atgaacggga atttgctact aaaatggaat ttcccagtca ctggaccgta caaatgccta
     1801 acgctcggcc ttttatcgat gcctatcgtc ggcgtcccga catgaacccc gtggttctgg
     1861 aactcgccat tctcgatacc aatatcgtgc aagctcagtt tcaagaagaa ttgaaggaga
     1921 cctcccgttg gtgggaaagc acggggattg ttcaagaact gccgtttgtt cgggaccgga
     1981 ttgtggaagg ttatttttgg accattggtg ttactcaacg ccgtgaacac ggttacgaac
     2041 gtattatgac ggccaaagtc atcgctttgg tgacctgttt ggatgatatt tatgacgtat
     2101 atggcactat tgaagaattg caactcttca cctctacgat tcagcgttgg gatttggagt
     2161 ctatgaagca gttaccgact tatatgcagg taagcttcct ggccttgcac aattttgtaa
     2221 ccgaagtggc ctatgatacg ctgaagaaaa agggctacaa ctctaccccc tatttgcgga
     2281 agacttgggt ggatttggtc gaaagttaca ttaaggaagc cacttggtac tataatgggt
     2341 acaaaccctc tatgcaggaa tacctcaaca acgcctggat ctctgtgggc agcatggcta
     2401 ttttgaatca tttgtttttt cgctttacta atgaacgcat gcataagtac cgggacatga
     2461 atcgtgtatc ctctaatatt gtgcggttag ccgacgatat gggaacctct ttggccgaag
     2521 ttgaacgcgg tgacgtgccc aaagctatcc aatgttacat gaatgaaacg aacgcctctg
     2581 aggaggaggc ccgcgaatat gtgcggcgcg ttatccagga agaatgggaa aaactgaaca
     2641 ctgaactgat gcgcgacgac gacgatgacg atgatttcac cttaagtaaa tactactgcg
     2701 aagtcgttgc taacctgacc cggatggctc agttcattta ccaagatggt tccgatgggt
     2761 ttgggatgaa agattccaaa gtaaatcgtt tactgaaaga aacgctgatt gagcgctatg
     2821 agtgaagatc tgcggccgcg ttgatcggca cgtaagaggt tccaactttc accataatga
     2881 aataagatca ctaccgggcg tattttttga gttatcgaga ttttcaggag ctaaggaagc
     2941 taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa
     3001 agaacatttt gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct
     3061 ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt
     3121 tattcacatt cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga
     3181 cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac
     3241 tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat
     3301 atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat
     3361 tgagaatatg tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa
     3421 cgtggccaat atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca
     3481 aggcgacaag gtgctgatgc cgctggcgat tcaggttcat catgccgtct gtgatggctt
     3541 ccatgtcggc agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc
     3601 gtaatttttt taaggcagtt attggtgccc ttaaacgcct ggggatcctt ggtgtaatgc
     3661 caactgaata atctgcaaat tgcactctcc ttcaatgggg ggtgcttttt gcttgactga
     3721 gtaatcttct gattgctgat cttgattgcc atcgatcgcc ggggagtccg gggcagttac
     3781 cattagagag tctagagaat taatccatct tcgatagagg aattatgggg gaagaacctg
     3841 tgccggcgga taaagcatta ggcaagaaat tcaagaaaaa aaatgcctcc tggagcattg
     3901 aagaaagcga agctctgtac cgggttgagg cctggggggc accttatttt gccattaatg
     3961 ccgctggtaa cataaccgtc tctcccaacg gcgatcgggg cggttcgtta gatttgttgg
     4021 aactggtgga agccctgcgg caaagaaagc tcggcttacc cctattaatt cgtttttccg
     4081 atattttggc cgatcgccta gagcgattga atagttgttt tgccaaggcg atcgagctcc
     4141 agcttttgtt ccctttagtg agggttaatt tcgagcttgg cgtaatcatg gtcatagctg
     4201 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4261 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4321 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4381 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4441 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4501 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4561 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4621 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4681 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4741 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4801 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4861 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4921 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4981 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     5041 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5101 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5161 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5221 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5281 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5341 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5401 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5461 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5521 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5581 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5641 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5701 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5761 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5821 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5881 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5941 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6001 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6061 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6121 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6181 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6241 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6301 caaatagggg ttccgcgcac atttccccga aaagtgc
//
