LOCUS       sequence-479072-        2762 bp    DNA     circular SYN 22-JUL-2025
DEFINITION  Expresses C-terminal 6xHis-tagged CYP168A1 in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2762)
  TITLE     Pseudomonas aeruginosa CYP168A1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 2762)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 22, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pET28a-CYP168A1
FEATURES             Location/Qualifiers
     source          1..2762
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(64..111)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     107..125
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(178..195)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(284..306)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    392..531
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     447..466
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(546..563)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(717..1305)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(797..816)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1416..1435)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1427..2242
                     /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"
     CDS             1427..2242
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1500..1519)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(join(2334..2762,1..27))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2471..2492)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2683..2702
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgcttaatgc gccgctacag ggcgcgtccc attcgccaat ccggatatag ttcctccttt
       61 cagcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta gttattgctc
      121 agcggtggca gcagccaact cagcttcctt tcgggctttg ttagcagccg gatctcagtg
      181 gtggtggtgg tggtgctcga gtgcggccgc atgaatcgct tcacgaccac gctgatgagc
      241 tttaccgcag ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
      301 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
      361 gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata
      421 gcggagtgta tactggctta actatgcggc atcagagcag attgtactga gagtgcacca
      481 ttgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc
      541 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
      601 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
      661 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      721 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      781 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      841 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      901 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      961 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     1021 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     1081 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     1141 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     1201 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     1261 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     1321 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     1381 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     1441 acgggaaacg tcttgctcta ggccgcgatt aaattccaac atggatgctg atttatatgg
     1501 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gattgtatgg
     1561 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     1621 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     1681 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccgggaaaac
     1741 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     1801 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     1861 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     1921 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     1981 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     2041 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     2101 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     2161 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     2221 gatgctcgat gagtttttct aagaattaat tcatgagcgg atacatattt gaatgtattt
     2281 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt
     2341 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     2401 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     2461 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     2521 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     2581 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     2641 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     2701 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     2761 cg
//
