LOCUS       V014016                 8326 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014016
VERSION     V014016
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8326)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8326
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        494..598
                     /label="AmpR promoter"
     CDS             599..1456
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      1630..2218
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(2404..2546)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(2955..3128)
                     /label="lacZ-alpha"
                     /note="LacZ-alpha fragment of beta-galactosidase"
     protein_bind    complement(3148..3164)
                     /label="lac operator"
                     /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(3172..3202)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3217..3238)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3254..4333)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(4334..4411)
                     /label="lacI promoter"
     promoter        4529..4557
                     /label="tac promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    4565..4581
                     /label="lac operator"
                     /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             4727..6157
                     /gene="CYP8B1"
                     /label="7-alpha-hydroxycholest-4-en-3-one
                     12-alpha-hydroxylase"
                     /note="7-alpha-hydroxycholest-4-en-3-one
                     12-alpha-hydroxylase from Homo sapiens. Accession#: Q9UNU6"
     CDS             6158..6175
                     /label="6xHis"
                     /note="6xHis affinity tag"
     protein_bind    complement(6188..6221)
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        6236..6681
                     /label="sacB promoter"
                     /note="sacB promoter and control region"
     CDS             6682..8100
                     /label="SacB"
                     /note="secreted levansucrase that renders bacterial growth
                     sensitive to sucrose"
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt
      481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      541 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
      601 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
      661 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
      721 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
      781 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
      841 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
      901 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
      961 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     1021 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     1081 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     1141 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     1201 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     1261 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     1321 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     1381 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     1441 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     1501 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1561 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1621 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1681 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1741 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     1801 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1861 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1921 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1981 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2041 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2101 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2161 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2221 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     2281 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     2341 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     2401 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatatgg
     2461 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     2521 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     2581 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     2641 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagaac gccatcaaaa
     2701 ataattcgcg tctggccttc ctgtagccag ctttcatcaa cattaaatgt gagcgagtaa
     2761 caacccgtcg gattctccgt gggaacaaac ggcggattga ccgtaatggg ataggttacg
     2821 ttggtgtaga tgggcgcatc gtaaccgtgc atctgccagt ttgaggggac gacgacagta
     2881 tcggcctcag gaagatcgca ctccagccag ctttccggca ccgcttctgg tgccggaaac
     2941 caggcaaagc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     3001 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     3061 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga atccgtaatc
     3121 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     3181 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     3241 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     3301 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
     3361 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
     3421 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
     3481 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac
     3541 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
     3601 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
     3661 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
     3721 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
     3781 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
     3841 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
     3901 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
     3961 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
     4021 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
     4081 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
     4141 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
     4201 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
     4261 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
     4321 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     4381 cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac
     4441 tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg
     4501 aatggtgcat gctcacgagg tcctcgagtt gacaattaat catcggctcg tataatgtgt
     4561 ggaattgtga gcggataaca atttcacaca ggaaacagga tcagcttact ccccatcccc
     4621 ctgggatcca tcgatgcttt tgacaattaa tcatcggctc gtataatgtg tggaattgtg
     4681 agcggataac aaaggaggtc atatgatggc caagaagacc agcagcctcc gacaacgcag
     4741 gccatgggag ccccctctgg acaagggtac cgtgccctgg cttggccatg ccatggcttt
     4801 ccggaagaat atgtttgaat ttctgaagcg catgaggacc aagcatgggg atgtgttcac
     4861 agtgcagcta gggggccagt acttcacctt cgtcatggac cccctctcct ttggctccat
     4921 cctcaaggac acacagagaa aactagactt tgggcaatat gcaaaaaaac tggtgctgaa
     4981 ggtatttgga taccgttcag tgcaagggga ccatgagatg atacactcag ccagcaccaa
     5041 gcatctgagg ggggatggct tgaaggatct taatgagacc atgctggaca gcctgtcctt
     5101 tgtaatgctg acgtccaaag gctggagtct ggatgccagt tgctggcatg aggacagcct
     5161 ctttcgcttc tgctattaca tcttgttcac agctggctac ctgagcttgt tcggctacac
     5221 gaaggacaag gagcaggacc tgctacaggc aggagagtta ttcatggagt tccgcaagtt
     5281 tgaccttctt ttcccaaggt ttgtctactc cctgctgtgg ccccgggagt ggctagaagt
     5341 gggccgactc cagcgtctct ttcacaagat gctctccgtg agccacagcc aggagaagga
     5401 gggcatcagc aactggctgg gcaacatgct tcagtttctg agggagcagg gggtaccctc
     5461 agctatgcag gacaagttca acttcatgat gctctgggcc tcccagggga acacggggcc
     5521 tacctctttc tgggccctct tgtacctcct gaagcaccca gaagctattc gggctgtgag
     5581 ggaggaagct acccaggtcc tgggtgaggc caggctggag accaagcagt cctttgcctt
     5641 caaactcggt gccctgcaac acaccccagt tctagacagc gtggtggagg agacgctgcg
     5701 gctgagggct gcacccaccc tcctcaggtt ggttcatgaa gactataccc tgaagatgtc
     5761 cagtgggcag gagtatctgt tccgccatgg agacatcctg gccctctttc cctacctctc
     5821 agtgcacatg gaccctgaca tccaccctga gcccaccgtc ttcaagtacg atcgcttcct
     5881 caaccctaat ggcagccgga aagtggactt cttcaagaca ggcaagaaga tccaccacta
     5941 caccatgccc tggggttcgg gcgtttccat ctgccctggg aggttctttg cactcagtga
     6001 ggtgaagctc tttatcctgc ttatggtcac acactttgac ttagagttgg tggaccctga
     6061 cacaccacta ccccatgttg acccgcagcg ctggggtttt ggcaccatgc agcccagcca
     6121 cgatgtgcgc ttccgctacc gcctgcatcc tacagagcac catcaccatc accattagga
     6181 aagatccata acttcgtata gcatacatta tacgaagtta tgcggccgcg acgtccacat
     6241 atacctgccg ttcactatta tttagtgaaa tgagatatta tgatattttc tgaattgtga
     6301 ttaaaaaggc aactttatgc ccatgcaaca gaaactataa aaaatacaga gaatgaaaag
     6361 aaacagatag attttttagt tctttaggcc cgtagtctgc aaatcctttt atgattttct
     6421 atcaaacaaa agaggaaaat agaccagttg caatccaaac gagagtctaa tagaatgagg
     6481 tcgaaaagta aatcgcgcgg gtttgttact gataaagcag gcaagaccta aaatgtgtaa
     6541 agggcaaagt gtatactttg gcgtcacccc ttacatattt taggtctttt tttattgtgc
     6601 gtaactaact tgccatcttc aaacaggagg gctggaagaa gcagaccgct aacacagtac
     6661 ataaaaaagg agacatgaac gatgaacatc aaaaagtttg caaaacaagc aacagtatta
     6721 acctttacta ccgcactgct ggcaggaggc gcaactcaag cgtttgcgaa agaaacgaac
     6781 caaaagccat ataaggaaac atacggcatt tcccatatta cacgccatga tatgctgcaa
     6841 atccctgaac agcaaaaaaa tgaaaaatat caagttcctg agttcgattc gtccacaatt
     6901 aaaaatatct cttctgcaaa aggcctggac gtttgggaca gctggccatt acaaaacgct
     6961 gacggcactg tcgcaaacta tcacggctac cacatcgtct ttgcattagc cggagatcct
     7021 aaaaatgcgg atgacacatc gatttacatg ttctatcaaa aagtcggcga aacttctatt
     7081 gacagctgga aaaacgctgg ccgcgtcttt aaagacagcg acaaattcga tgcaaatgat
     7141 tctatcctaa aagaccaaac acaagaatgg tcaggttcag ccacatttac atctgacgga
     7201 aaaatccgtt tattctacac tgatttctcc ggtaaacatt acggcaaaca aacactgaca
     7261 actgcacaag ttaacgtatc agcatcagac agctctttga acatcaacgg tgtagaggat
     7321 tataaatcaa tctttgacgg tgacggaaaa acgtatcaaa atgtacagca gttcatcgat
     7381 gaaggcaact acagctcagg cgacaaccat acgctgagag atcctcacta cgtagaagat
     7441 aaaggccaca aatacttagt atttgaagca aacactggaa ctgaagatgg ctaccaaggc
     7501 gaagaatctt tatttaacaa agcatactat ggcaaaagca catcattctt ccgtcaagaa
     7561 agtcaaaaac ttctgcaaag cgataaaaaa cgcacggctg agttagcaaa cggcgctctc
     7621 ggtatgattg agctaaacga tgattacaca ctgaaaaaag tgatgaaacc gctgattgca
     7681 tctaacacag taacagatga aattgaacgc gcgaacgtct ttaaaatgaa cggcaaatgg
     7741 tacctgttca ctgactcccg cggatcaaaa atgacgattg acggcattac gtctaacgat
     7801 atttacatgc ttggttatgt ttctaattct ttaactggcc catacaagcc gctgaacaaa
     7861 actggccttg tgttaaaaat ggatcttgat cctaacgatg taacctttac ttactcacac
     7921 ttcgctgtac ctcaagcgaa aggaaacaat gtcgtgatta caagctatat gacaaacaga
     7981 ggattctacg cagacaaaca atcaacgttt gcgccgagct tcctgctgaa catcaaaggc
     8041 aagaaaacat ctgttgtcaa agacagcatc cttgaacaag gacaattaac agttaacaaa
     8101 taaaaacgca aaagaaaatg ccgatattga cgtcaggtgg cacttttcga ggagatcatg
     8161 cacatgatga cgaagcttat cgatgataag ctgtcaaaca tgagcagatc tgagcccgcc
     8221 taatgagcgg gcttttagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     8281 catgataata atggtttctt aggtgctcgt caaagcaact ccggat
//
