LOCUS       sequence-368445-        6769 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal His-tagged CYP110D1 coding sequence under the control of 
            PpsbA2* promoter and the BBa_B0032 RBS, for the expression in 
            Synechocystis..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6769)
  AUTHORS   Mascia F, Pereira SB, Pacheco CC, Oliveira P, Solarczek J, Schallmey
            A, Kourist R, Alphand V, Tamagnini P.
  TITLE     Light-driven hydroxylation of testosterone by Synechocystis sp. PCC 
            6803 expressing the heterologous CYP450 monooxygenase CYP110D1
  JOURNAL   Green Chemistry, 24 (2022):6156
REFERENCE   2  (bases 1 to 6769)
  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"; journalName: "Green 
            Chemistry, 24 (2022):6156"
COMMENT     Sequence Label: pSEVA251::PpsbA2*::B0032::CYP110D1 N-terminal
            ...
FEATURES             Location/Qualifiers
     source          1..6769
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     9..31
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    121..142
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             226..243
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1663..1757
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1878..1897)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1889..2704
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            2871..2979
                     /note="incP origin of transfer"
     CDS             complement(2990..3841)
                     /codon_start=1
                     /gene="repC"
                     /product="replication protein C of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepC
                     /translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TADAMVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     CDS             complement(5167..6138)
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepB
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     rep_origin      6245..6639
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     terminator      complement(6677..6763)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaattaaag cggataacaa tttcacacag gaggccgcct aggccgcggc cgcgcgaatt
       61 cgagctcggt acccgggcta attagggggt gtcgcccttt acacgtactt agtcgctgaa
      121 gaattcgcgg ccgcttctag agagctttac aaaactctca ttaatccttt agactaagtt
      181 tagtcagttc caatctgtac tagtcacaca ggaaagtact agatgcacca tcaccatcac
      241 catacagtca ctcaaaacct acccaacggc cccagaatac cccggttgtt acggttgttt
      301 aagtttatta ctcaaccaat acagtatgtg gaagattttg ccaaagtcta cggtgacaac
      361 ttcactatct gggggagtgg agaaagttat tttgtctact ttagccatcc ccaagcattg
      421 gaacagattt ttactaatgt tagttgtttt gagagtagtg gtggaggtag tccgctttta
      481 gaacttttat tgggtaagaa ttctttaatt ttactagaag gcgatcgcca tcagcgccaa
      541 cgtcaactat taaccccacc tttccacggt gaaaggatgc gggcttatgg tcaaactatt
      601 cgtgaaataa ctcaacaagt cactcaagca tggcaaatgg gtaagccctt caatatccgg
      661 gcttccatgc aggaaattac catgcgtgtc attttgcggg tagtgtttgg tgttgatgag
      721 ggggaattgt ttcaagaact gcggcaattg ctaactaccc tgttagattt catgggttct
      781 cctctaatgt ccagcacctt cttttttagt ttcacgcaaa aggactatgg tgcatggagt
      841 ccttgggggc gaatggtgcg cctcatcaaa aaaattgacc aactgattta cgctttgatt
      901 gctcaacgtc gggctgagtt tggcgaaaat cgtcaagata tccttagttt attaatttcg
      961 gcacgttatg acgacggaca gccaatgtca gacgtagagt tacgggatga attaatgaca
     1021 atgttggttg caggacatga aaccactgct tcagccttga cttgggcgtt ttattggatt
     1081 gatagcgtac cagaagtacg tgaaaagtta tttcaggaat tagatacact gaatgacgat
     1141 tcagaaccca gtattattgc caaattgcct tatttaacag cagtttgcca agaaacccta
     1201 cgcttttatc ctattgtgct gaatgctttc ttccggcgaa caaaaaaccc tatggaaatt
     1261 atgggctaca aactgccgaa agcaacatta gttgttccta gtatttattt agcccatcat
     1321 cgagaagaag tttatccgca atccaagcaa ttccgaccag aacgcttttt agaaaagcaa
     1381 ttctcccctt acgaatactt accctttggc ggcggtaatc ggcgctgcat cggtctagca
     1441 tttgcccaat atgaaatgaa aattgtctta gcaacaattt tgtcacaatt ccaagtatct
     1501 agactcagta agcgtccggt gcagcctgtg cgtcggggat taactttggc agcaccagga
     1561 ggaatgaaga tggtggctaa taaaagaatg cgtaattcgt aactgcaggc atgcaagctt
     1621 gcggccgcgt cgtgactggg aaaaccctgg cgactagtct tggactcctg ttgatagatc
     1681 cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc cgccgggcgt
     1741 tttttattgg tgagaatcca ggggtcccca ataattacga tttaaatttg tgtctcaaaa
     1801 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     1861 ttacataaac agtaatacaa ggggtgttat gagccatatt cagcgtgaaa cgagctgtag
     1921 ccgtccgcgt ctgaacagca acatggatgc ggatctgtat ggctataaat gggcgcgtga
     1981 taacgtgggt cagagcggcg cgaccattta tcgtctgtat ggcaaaccgg atgcgccgga
     2041 actgtttctg aaacatggca aaggcagcgt ggcgaacgat gtgaccgatg aaatggtgcg
     2101 tctgaactgg ctgaccgaat ttatgccgct gccgaccatt aaacatttta ttcgcacccc
     2161 ggatgatgcg tggctgctga ccaccgcgat tccgggcaaa accgcgtttc aggtgctgga
     2221 agaatatccg gatagcggcg aaaacattgt ggatgcgctg gccgtgtttc tgcgtcgtct
     2281 gcatagcatt ccggtgtgca actgcccgtt taacagcgat cgtgtgtttc gtctggccca
     2341 ggcgcagagc cgtatgaaca acggcctggt ggatgcgagc gattttgatg atgaacgtaa
     2401 cggctggccg gtggaacagg tgtggaaaga aatgcataaa ctgctgccgt ttagcccgga
     2461 tagcgtggtg acccacggcg attttagcct ggataacctg attttcgatg aaggcaaact
     2521 gattggctgc attgatgtgg gccgtgtggg cattgcggat cgttatcagg atctggccat
     2581 tctgtggaac tgcctgggcg aatttagccc gagcctgcaa aaacgtctgt ttcagaaata
     2641 tggcattgat aatccggata tgaacaaact gcaatttcat ctgatgctgg atgaattttt
     2701 ctaataatta attggaccgc ggtccgcgcg ttgtcctttt ccgctgcata accctgcttc
     2761 ggggtcatta tagcgatttt ttcggtatat ccatcctttt tcgcacgata tacaggattt
     2821 tgccaaaggg ttcgtgtaga ctttccttgg tgtatccaac ggcgtcagcc gggcaggata
     2881 ggtgaagtag gcccacccgc gagcgggtgt tccttcttca ctgtccctta ttcgcacctg
     2941 gcggtgctca acgggaatcc tgctctgcga ggctggccgt aggccggcct cagcctgccg
     3001 ccttgggccg ggtgatgtcg tacttgcccg ccgcgaactc ggttaccgtc cagcccagcg
     3061 cgaccagctc cggcaacgcc tcgcgcaccc gctggcggcg cttgcgcatg gtcgaaccac
     3121 tggcctctga cggccagaca tagccgcaca aggtatctat ggaagccttg ccggttttgc
     3181 cggggtcgat ccagccacac agccgctggt gcagcaggcg ggcggtttcg ctgtccagcg
     3241 cccgcacctc gtccatgctg atgcgcacat gctggccgcc acccatgacg gcctgcgcga
     3301 tcaaggggtt cagggccacg tacaggcgcc cgtccgcctc gtcgctggcg tactccgaca
     3361 gcagccgaaa cccctgccgc ttgcggccat tctgggcgat gatggatacc ttccaaaggc
     3421 gctcgatgca gtcctgtatg tgcttgagcg ccccaccact atcgacctct gccccgattt
     3481 cctttgccag cgcccgatag ctacctttga ccaccatggc atcagcggtg acggcctccc
     3541 acttgggttc caggaacagc cggagctgcc gtccgccttc ggtcttgggt tccgggccaa
     3601 gcactaggcc attaggccca gccatggcca ccagcccttg caggatgcgc agatcatcag
     3661 cgcccagcgg ctccgggccg ctgaactcga tccgcttgcc gtcgccgtag tcatacgtca
     3721 cgtccagctt gctgcgcttg cgctcgcccc gcttgagggc acggaacagg ccgggggcca
     3781 gacagtgcgc cgggtcgtgc cggacgtggc tgaggctgtg cttgttctta ggcttcacca
     3841 cggggcaccc ccttgctctt gcgctgcctc tccagcacgg cgggcttgag caccccgccg
     3901 tcatgccgcc tgaaccaccg atcagcgaac ggtgcgccat agttggcctt gctcacaccg
     3961 aagcggacga agaaccggcg ctggtcgtcg tccacacccc attcctcggc ctcggcgctg
     4021 gtcatgctcg acaggtagga ctgccagcgg atgttatcga ccagtaccga gctgccccgg
     4081 ctggcctgct gctggtcgcc tgcgcccatc atggccgcgc ccttgctggc atggtgcagg
     4141 aacacgatag agcacccggt atcggcggcg atggcctcca tgcgaccgat gacctgggcc
     4201 atggggccgc tggcgttttc ttcctcgatg tggaaccggc gcagcgtgtc cagcaccatc
     4261 aggcggcggc cctcggcggc gcgcttgagg ccgtcgaacc actccggggc catgatgttg
     4321 ggcaggctgc cgatcagcgg ctggatcagc aggccgtcag ccacggcttg ccgttcctcg
     4381 gcgctgaggt gcgccccaag ggcgtgcagg cggggtgggc gggtcttcgg cgggcaggta
     4441 gatcaccggg ccggtgggca gttcgcccac ctccagcaga tccggcccgc ctgcaatctg
     4501 tgcggccagt tgcagggcca gcatggattt accggcacca ccgggcgaca ccagcgcccc
     4561 gaccgtaccg gccaccatgt tgggcaaaac gtagtccagc ggtggcggcg ctgctgcgaa
     4621 cgcctccaga atattgatag gcttatgggt agccattgat tgcctccttt gcaggcagtt
     4681 ggtggttagg cgctggcggg gtcactaccc ccgccctgcg ccgctctgag ttcttccagg
     4741 cactcgcgca gcgcctcgta ttcgtcgtcg gtcagccaga acttgcgctg acgcatccct
     4801 ttggccttca tgcgctcggc atatcgcgct tggcgtacag cgtcagggct ggccagcagg
     4861 tcgccggtct gcttgtcctt ttggtctttc atatcagtca ccgagaaact tgccggggcc
     4921 gaaaggcttg tcttcgcgga acaaggacaa ggtgcagccg tcaaggttaa ggctggccat
     4981 atcagcgact gaaaagcggc cagcctcggc cttgtttgac gtataaccaa agccaccggg
     5041 caaccaatag cccttgtcac ttttgatcag gtagaccgac cctgaagcgc ttttttcgta
     5101 ttccataaaa cccccttctg tgcgtgagta ctcatagtat aacaggcgtg agtaccaacg
     5161 caagcactac atgctgaaat ctggcccgcc cctgtccatg cctcgctggc ggggtgccgg
     5221 tgcccgtgcc agctcggccc gcgcaagctg gacgctgggc agacccatga ccttgctgac
     5281 ggtgcgctcg atgtaatccg cttcgtggcc gggcttgcgc tctgccagcg ctgggctggc
     5341 ctcggccatg gccttgccga tttcctcggc actgcggccc cggctggcca gcttctgcgc
     5401 ggcgataaag tcgcacttgc tgaggtcatc accgaagcgc ttgaccagcc cggccatctc
     5461 gctgcggtac tcgtccagcg ccgtgcgccg gtggcggcta agctgccgct cgggcagttc
     5521 gaggctggcc agcctgcggg ccttctcctg ctgccgctgg gcctgctcga tctgctggcc
     5581 agcctgctgc accagcgccg ggccagcggt ggcggtcttg cccttggatt cacgcagcag
     5641 cacccacggc tgataaccgg cgcgggtggt gtgcttgtcc ttgcggttgg tgaagcccgc
     5701 caagcggcca tagtggcggc tgtcggcgct ggccgggtcg gcgtcgtact cgctggccag
     5761 cgtccgggca atctgccccc gaagttcacc gcctgcggcg tcggccacct tgacccatgc
     5821 ctgatagttc ttcgggctgg tttccactac cagggcaggc tcccggccct cggctttcat
     5881 gtcatccagg tcaaactcgc tgaggtcgtc caccagcacc agaccatgcc gctcctgctc
     5941 ggcgggcctg atatacacgt cattgccctg ggcattcatc cgcttgagcc atggcgtgtt
     6001 ctggagcact tcggcggctg accattcccg gttcatcatc tggccggtgg tggcgtccct
     6061 gacgccgata tcgaagcgct cacagcccat ggccttgagc tgtcggccta tggcctgcaa
     6121 agtcctgtcg ttcttcatcg ggccaccaag cgattcccac acattatacg agccggaagc
     6181 ataaagtgta gctagatccg aaggatgagc cgggctgaat gatcgaccga gacaggccct
     6241 gcggggctgc acacgcgccc ccacccttcg ggtaggggga aaggccgcta aagcggctaa
     6301 aagcgctcca gcgtatttct gcggggtttg gtgtggggtt tagcgggctt tgcccgcctt
     6361 tccccctgcc gcgcagcggt ggggcggtgt gtagcctagc gcagcgaata gaccagctat
     6421 ccggcctctg gccgggcata ttgggcaagg gcagcagcgc cccacaaggg cgctgataac
     6481 cgcgcctagt ggattattct tagataatca tggatggatt tttccaacac cccgccagcc
     6541 cccgcccctg ctgggtttgc aggtttgggg gcgtgacagt tattgcaggg gttcgtgaca
     6601 gttattgcag gggggcgtga cagttattgc aggggttcgt gacagttagg gcgcgcccag
     6661 ctgtctaggg cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa
     6721 aacgaaaggc ccagtctttc gactgagcct ttcgttttat ttgatgcct
//
