LOCUS       sequence-368444-        6774 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal His-tagged CYP110D1 coding sequence under the control of 
            Ptrc.x.tetO2 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 6774)
  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 6774)
  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::Ptrc.x.tetO2::B0032::CYP110D1 N-term...
FEATURES             Location/Qualifiers
     source          1..6774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     9..31
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    101..122
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        join(129..134,135..151,152..158)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             220..237
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1657..1751
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1872..1891)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1883..2698
                     /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            2865..2973
                     /note="incP origin of transfer"
     CDS             complement(2984..3835)
                     /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(3822..4661)
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepA
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     CDS             complement(5172..6143)
                     /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      6250..6644
                     /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(6682..6768)
                     /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 gaattcgcgg ccgcttctag
      121 aggagctgtt gacaattaat catccggctc gtataatgtg tggatccccg tcagtgacgg
      181 agattcacac atactagtca cacaggaaag tactagatgc accatcacca tcaccataca
      241 gtcactcaaa acctacccaa cggccccaga ataccccggt tgttacggtt gtttaagttt
      301 attactcaac caatacagta tgtggaagat tttgccaaag tctacggtga caacttcact
      361 atctggggga gtggagaaag ttattttgtc tactttagcc atccccaagc attggaacag
      421 atttttacta atgttagttg ttttgagagt agtggtggag gtagtccgct tttagaactt
      481 ttattgggta agaattcttt aattttacta gaaggcgatc gccatcagcg ccaacgtcaa
      541 ctattaaccc cacctttcca cggtgaaagg atgcgggctt atggtcaaac tattcgtgaa
      601 ataactcaac aagtcactca agcatggcaa atgggtaagc ccttcaatat ccgggcttcc
      661 atgcaggaaa ttaccatgcg tgtcattttg cgggtagtgt ttggtgttga tgagggggaa
      721 ttgtttcaag aactgcggca attgctaact accctgttag atttcatggg ttctcctcta
      781 atgtccagca ccttcttttt tagtttcacg caaaaggact atggtgcatg gagtccttgg
      841 gggcgaatgg tgcgcctcat caaaaaaatt gaccaactga tttacgcttt gattgctcaa
      901 cgtcgggctg agtttggcga aaatcgtcaa gatatcctta gtttattaat ttcggcacgt
      961 tatgacgacg gacagccaat gtcagacgta gagttacggg atgaattaat gacaatgttg
     1021 gttgcaggac atgaaaccac tgcttcagcc ttgacttggg cgttttattg gattgatagc
     1081 gtaccagaag tacgtgaaaa gttatttcag gaattagata cactgaatga cgattcagaa
     1141 cccagtatta ttgccaaatt gccttattta acagcagttt gccaagaaac cctacgcttt
     1201 tatcctattg tgctgaatgc tttcttccgg cgaacaaaaa accctatgga aattatgggc
     1261 tacaaactgc cgaaagcaac attagttgtt cctagtattt atttagccca tcatcgagaa
     1321 gaagtttatc cgcaatccaa gcaattccga ccagaacgct ttttagaaaa gcaattctcc
     1381 ccttacgaat acttaccctt tggcggcggt aatcggcgct gcatcggtct agcatttgcc
     1441 caatatgaaa tgaaaattgt cttagcaaca attttgtcac aattccaagt atctagactc
     1501 agtaagcgtc cggtgcagcc tgtgcgtcgg ggattaactt tggcagcacc aggaggaatg
     1561 aagatggtgg ctaataaaag aatgcgtaat tcgtaactgc aggcatgcaa gcttgcggcc
     1621 gcgtcgtgac tgggaaaacc ctggcgacta gtcttggact cctgttgata gatccagtaa
     1681 tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg gcgtttttta
     1741 ttggtgagaa tccaggggtc cccaataatt acgatttaaa tttgtgtctc aaaatctctg
     1801 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     1861 aaacagtaat acaaggggtg ttatgagcca tattcagcgt gaaacgagct gtagccgtcc
     1921 gcgtctgaac agcaacatgg atgcggatct gtatggctat aaatgggcgc gtgataacgt
     1981 gggtcagagc ggcgcgacca tttatcgtct gtatggcaaa ccggatgcgc cggaactgtt
     2041 tctgaaacat ggcaaaggca gcgtggcgaa cgatgtgacc gatgaaatgg tgcgtctgaa
     2101 ctggctgacc gaatttatgc cgctgccgac cattaaacat tttattcgca ccccggatga
     2161 tgcgtggctg ctgaccaccg cgattccggg caaaaccgcg tttcaggtgc tggaagaata
     2221 tccggatagc ggcgaaaaca ttgtggatgc gctggccgtg tttctgcgtc gtctgcatag
     2281 cattccggtg tgcaactgcc cgtttaacag cgatcgtgtg tttcgtctgg cccaggcgca
     2341 gagccgtatg aacaacggcc tggtggatgc gagcgatttt gatgatgaac gtaacggctg
     2401 gccggtggaa caggtgtgga aagaaatgca taaactgctg ccgtttagcc cggatagcgt
     2461 ggtgacccac ggcgatttta gcctggataa cctgattttc gatgaaggca aactgattgg
     2521 ctgcattgat gtgggccgtg tgggcattgc ggatcgttat caggatctgg ccattctgtg
     2581 gaactgcctg ggcgaattta gcccgagcct gcaaaaacgt ctgtttcaga aatatggcat
     2641 tgataatccg gatatgaaca aactgcaatt tcatctgatg ctggatgaat ttttctaata
     2701 attaattgga ccgcggtccg cgcgttgtcc ttttccgctg cataaccctg cttcggggtc
     2761 attatagcga ttttttcggt atatccatcc tttttcgcac gatatacagg attttgccaa
     2821 agggttcgtg tagactttcc ttggtgtatc caacggcgtc agccgggcag gataggtgaa
     2881 gtaggcccac ccgcgagcgg gtgttccttc ttcactgtcc cttattcgca cctggcggtg
     2941 ctcaacggga atcctgctct gcgaggctgg ccgtaggccg gcctcagcct gccgccttgg
     3001 gccgggtgat gtcgtacttg cccgccgcga actcggttac cgtccagccc agcgcgacca
     3061 gctccggcaa cgcctcgcgc acccgctggc ggcgcttgcg catggtcgaa ccactggcct
     3121 ctgacggcca gacatagccg cacaaggtat ctatggaagc cttgccggtt ttgccggggt
     3181 cgatccagcc acacagccgc tggtgcagca ggcgggcggt ttcgctgtcc agcgcccgca
     3241 cctcgtccat gctgatgcgc acatgctggc cgccacccat gacggcctgc gcgatcaagg
     3301 ggttcagggc cacgtacagg cgcccgtccg cctcgtcgct ggcgtactcc gacagcagcc
     3361 gaaacccctg ccgcttgcgg ccattctggg cgatgatgga taccttccaa aggcgctcga
     3421 tgcagtcctg tatgtgcttg agcgccccac cactatcgac ctctgccccg atttcctttg
     3481 ccagcgcccg atagctacct ttgaccacca tggcatcagc ggtgacggcc tcccacttgg
     3541 gttccaggaa cagccggagc tgccgtccgc cttcggtctt gggttccggg ccaagcacta
     3601 ggccattagg cccagccatg gccaccagcc cttgcaggat gcgcagatca tcagcgccca
     3661 gcggctccgg gccgctgaac tcgatccgct tgccgtcgcc gtagtcatac gtcacgtcca
     3721 gcttgctgcg cttgcgctcg ccccgcttga gggcacggaa caggccgggg gccagacagt
     3781 gcgccgggtc gtgccggacg tggctgaggc tgtgcttgtt cttaggcttc accacggggc
     3841 acccccttgc tcttgcgctg cctctccagc acggcgggct tgagcacccc gccgtcatgc
     3901 cgcctgaacc accgatcagc gaacggtgcg ccatagttgg ccttgctcac accgaagcgg
     3961 acgaagaacc ggcgctggtc gtcgtccaca ccccattcct cggcctcggc gctggtcatg
     4021 ctcgacaggt aggactgcca gcggatgtta tcgaccagta ccgagctgcc ccggctggcc
     4081 tgctgctggt cgcctgcgcc catcatggcc gcgcccttgc tggcatggtg caggaacacg
     4141 atagagcacc cggtatcggc ggcgatggcc tccatgcgac cgatgacctg ggccatgggg
     4201 ccgctggcgt tttcttcctc gatgtggaac cggcgcagcg tgtccagcac catcaggcgg
     4261 cggccctcgg cggcgcgctt gaggccgtcg aaccactccg gggccatgat gttgggcagg
     4321 ctgccgatca gcggctggat cagcaggccg tcagccacgg cttgccgttc ctcggcgctg
     4381 aggtgcgccc caagggcgtg caggcggtga tgaatggcgg tgggcgggtc ttcggcgggc
     4441 aggtagatca ccgggccggt gggcagttcg cccacctcca gcagatccgg cccgcctgca
     4501 atctgtgcgg ccagttgcag ggccagcatg gatttaccgg caccaccggg cgacaccagc
     4561 gccccgaccg taccggccac catgttgggc aaaacgtagt ccagcggtgg cggcgctgct
     4621 gcgaacgcct ccagaatatt gataggctta tgggtagcca ttgattgcct cctttgcagg
     4681 cagttggtgg ttaggcgctg gcggggtcac tacccccgcc ctgcgccgct ctgagttctt
     4741 ccaggcactc gcgcagcgcc tcgtattcgt cgtcggtcag ccagaacttg cgctgacgca
     4801 tccctttggc cttcatgcgc tcggcatatc gcgcttggcg tacagcgtca gggctggcca
     4861 gcaggtcgcc ggtctgcttg tccttttggt ctttcatatc agtcaccgag aaacttgccg
     4921 gggccgaaag gcttgtcttc gcggaacaag gacaaggtgc agccgtcaag gttaaggctg
     4981 gccatatcag cgactgaaaa gcggccagcc tcggccttgt ttgacgtata accaaagcca
     5041 ccgggcaacc aatagccctt gtcacttttg atcaggtaga ccgaccctga agcgcttttt
     5101 tcgtattcca taaaaccccc ttctgtgcgt gagtactcat agtataacag gcgtgagtac
     5161 caacgcaagc actacatgct gaaatctggc ccgcccctgt ccatgcctcg ctggcggggt
     5221 gccggtgccc gtgccagctc ggcccgcgca agctggacgc tgggcagacc catgaccttg
     5281 ctgacggtgc gctcgatgta atccgcttcg tggccgggct tgcgctctgc cagcgctggg
     5341 ctggcctcgg ccatggcctt gccgatttcc tcggcactgc ggccccggct ggccagcttc
     5401 tgcgcggcga taaagtcgca cttgctgagg tcatcaccga agcgcttgac cagcccggcc
     5461 atctcgctgc ggtactcgtc cagcgccgtg cgccggtggc ggctaagctg ccgctcgggc
     5521 agttcgaggc tggccagcct gcgggccttc tcctgctgcc gctgggcctg ctcgatctgc
     5581 tggccagcct gctgcaccag cgccgggcca gcggtggcgg tcttgccctt ggattcacgc
     5641 agcagcaccc acggctgata accggcgcgg gtggtgtgct tgtccttgcg gttggtgaag
     5701 cccgccaagc ggccatagtg gcggctgtcg gcgctggccg ggtcggcgtc gtactcgctg
     5761 gccagcgtcc gggcaatctg cccccgaagt tcaccgcctg cggcgtcggc caccttgacc
     5821 catgcctgat agttcttcgg gctggtttcc actaccaggg caggctcccg gccctcggct
     5881 ttcatgtcat ccaggtcaaa ctcgctgagg tcgtccacca gcaccagacc atgccgctcc
     5941 tgctcggcgg gcctgatata cacgtcattg ccctgggcat tcatccgctt gagccatggc
     6001 gtgttctgga gcacttcggc ggctgaccat tcccggttca tcatctggcc ggtggtggcg
     6061 tccctgacgc cgatatcgaa gcgctcacag cccatggcct tgagctgtcg gcctatggcc
     6121 tgcaaagtcc tgtcgttctt catcgggcca ccaagcgatt cccacacatt atacgagccg
     6181 gaagcataaa gtgtagctag atccgaagga tgagccgggc tgaatgatcg accgagacag
     6241 gccctgcggg gctgcacacg cgcccccacc cttcgggtag ggggaaaggc cgctaaagcg
     6301 gctaaaagcg ctccagcgta tttctgcggg gtttggtgtg gggtttagcg ggctttgccc
     6361 gcctttcccc ctgccgcgca gcggtggggc ggtgtgtagc ctagcgcagc gaatagacca
     6421 gctatccggc ctctggccgg gcatattggg caagggcagc agcgccccac aagggcgctg
     6481 ataaccgcgc ctagtggatt attcttagat aatcatggat ggatttttcc aacaccccgc
     6541 cagcccccgc ccctgctggg tttgcaggtt tgggggcgtg acagttattg caggggttcg
     6601 tgacagttat tgcagggggg cgtgacagtt attgcagggg ttcgtgacag ttagggcgcg
     6661 cccagctgtc tagggcggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
     6721 gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat gcct
//
