LOCUS       sequence-193146-        6455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For protein expression of CYP2C19.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6455)
  TITLE     Goldstein CYP2C9 and CYP2C9 in pOri+ for bacterial expression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6455)
  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: 
            "Unpublished"
COMMENT     Sequence Label: CYP2C19
FEATURES             Location/Qualifiers
     source          1..6455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(35..40,41..58,59..65)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    73..89
                     /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        join(144..149,150..165,166..172)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    180..196
                     /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        join(239..244,245..260,261..267)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    275..291
                     /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)."
     CDS             complement(1771..1782)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1918..1936)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      1998..2453
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2085..2104)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2295..2316
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2486..2590
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2591..2659,2660..3451)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2809..2828)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3622..4210
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4111..4130
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4364..4381
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4396..4537
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4463..4482)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4623..4645
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4658..5119)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVLPRAFR"
     primer_bind     5068..5090
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5082..5099
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5083..5099
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5115..5131)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5115..5131)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    5139..5155
                     /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(5163..5169,5170..5187,5188..5193))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             complement(5242..6324)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6286..6305
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6325..6402)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
ORIGIN
        1 atgtaagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
       61 taatgtgtgg aattgtgagc ggataacaat ttcacacagg aaacaggatc gatccatcga
      121 tgagcttact ccccatcccc ctgttgacaa ttaatcatcg gctcgtataa tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa caggatcagc ttactcccca tccccctgtt
      241 gacaattaat catcggctcg tataatgtgt ggaattgtga gcggataaca atttcacaca
      301 ggaaacagga tccatcgatg cttaggaggt catatggctc tgttattagc agtttttctc
      361 tgtctctcat gtttgcttct cctttcaatc tggagacaga gctctgggag aggaaaactc
      421 cctcctggcc ccactcctct cccagtgatt ggaaatatcc tacagataga tattaaggat
      481 gtcagcaaat ccttaaccaa tctctcaaaa atctatggcc ctgtgttcac tctgtatttt
      541 ggcctggaac gcatggtggt gctgcatgga tatgaagtgg tgaaggaagc cctgattgat
      601 cttggagagg agttttctgg aagaggccat ttcccactgg ctgaaagagc taacagagga
      661 tttggaatcg ttttcagcaa tggaaagaga tggaaggaga tccggcgttt ctccctcatg
      721 acgctgcgga attttgggat ggggaagagg agcattgagg accgtgttca agaggaagcc
      781 cgctgccttg tggaggagtt gagaaaaacc aaggcttcac cctgtgatcc cactttcatc
      841 ctgggctgtg ctccctgcaa tgtgatctgc tccattattt tccagaaacg tttcgattat
      901 aaagatcagc aatttcttaa cttgatggaa aaattgaatg aaaacatcag gattgtaagc
      961 accccctgga tccagatatg caataatttt cccactatca ttgattattt cccgggaacc
     1021 cataacaaat tacttaaaaa ccttgctttt atggaaagtg atattttgga gaaagtaaaa
     1081 gaacaccaag aatcgatgga catcaacaac cctcgggact ttattgattg cttcctgatc
     1141 aaaatggaga aggaaaagca aaaccaacag tctgaattca ctattgaaaa cttggtaatc
     1201 actgcagctg acttacttgg agctgggaca gagacaacaa gcacaaccct gagatatgct
     1261 ctccttctcc tgctgaagca cccagaggtc acagctaaag tccaggaaga gattgaacgt
     1321 gtcgttggca gaaaccggag cccctgcatg caggacaggg gccacatgcc ctacacagat
     1381 gctgtggtgc acgaggtcca gagatacatc gacctcatcc ccaccagcct gccccatgca
     1441 gtgacctgtg acgttaaatt cagaaactac ctcattccca agggcacaac catattaact
     1501 tccctcactt ctgtgctaca tgacaacaaa gaatttccca acccagagat gtttgaccct
     1561 cgtcactttc tggatgaagg tggaaatttt aagaaaagta actacttcat gcctttctca
     1621 gcaggaaaac ggatttgtgt gggagagggc ctggcccgca tggagctgtt tttattcctg
     1681 accttcattt tgcagaactt taacctgaaa tctctgattg acccaaagga ccttgacaca
     1741 actcctgttg tcaatggatt tgcttctgtc ccgcccttct atcagctgtg cttcattcct
     1801 gtctgaagaa gcacagatgg tctggcaagc ttatcgatga taagctgtca aacatgagca
     1861 gatctgagcc cgcctaatga gcgggctttt ttttcagatc tgcttgaaga cgaaagggcc
     1921 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgatgc
     1981 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2041 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2101 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     2161 tttagggttc cgatttagag ctttacggca cctcgacccc aaaaaacttg atttgggtga
     2221 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     2281 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggg
     2341 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     2401 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa tttcatcgtc
     2461 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2521 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     2581 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     2641 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     2701 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     2761 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     2821 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     2881 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     2941 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3001 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3061 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3121 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3181 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     3241 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     3301 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     3361 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     3421 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     3481 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3541 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3601 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3661 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3721 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     3781 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3841 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3901 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3961 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4021 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4081 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4141 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4201 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4261 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     4321 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     4381 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4441 ccgcataatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     4501 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     4561 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     4621 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagaa
     4681 cgccatcaaa aataattcgc gtctggcctt cctgtagcca gctttcatca acattaaatg
     4741 tgagcgagta acaacccgtc ggattctccg tgggaacaaa cggcggattg accgtaatgg
     4801 gataggtcac gttggtgtag atgggcgcat cgtaaccgtg catctgccag tttgagggga
     4861 cgacgacagt atcggcctca ggaagatcgc actccagcca gctttccggc accgcttctg
     4921 gtgccggaaa ccaggcaaag cgccattcgc cattcaggct gcgcaactgt tgggaagggc
     4981 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     5041 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     5101 aatccgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5161 cacattatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5221 cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5281 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt
     5341 ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg
     5401 agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat
     5461 ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga
     5521 gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat
     5581 ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt
     5641 ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg
     5701 attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa
     5761 tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag
     5821 tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc
     5881 aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc
     5941 cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc
     6001 tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg
     6061 atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga
     6121 ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg
     6181 aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg
     6241 gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac
     6301 atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta
     6361 tcatgccata ccgcgaaagg ttttgcacca ttcgatggtg tcctggcacg acaggtttcc
     6421 cgactggaaa gcgggcagtg agcgcaacgc aatta
//
