LOCUS       V024787                 7711 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024787
VERSION     V024787
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             775..2367
                     /label="CYP27A1(NM_000784)"
                     /note="CYP27A1(NM_000784)"
                     /gene="CYP27A1"
     CDS             2368..2385
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     misc_feature    2487..3075
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3141..3189
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3391..3819
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3833..4162
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4229..5023
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5199..5320
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5369..5385)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5393..5409
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(5417..5447)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5462..5483
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5771..6359)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6530..7390)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(7391..7495)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
      661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
      721 actctagagc tagcgtttaa acttaagctt ggtaccgagc tcggatccgc caccatggct
      781 gcgctgggct gcgcgaggct gaggtgggcg ctgcgagggg ccggccgtgg cctctgcccc
      841 cacggggcca gagccaaggc cgcgatccct gccgccctcc cctcggacaa ggccaccgga
      901 gctcccggag ccgggcctgg tgtccggcgg cggcaacgga gcttagagga gattccacgt
      961 ctaggacagc tgcgcttctt ctttcagctg ttcgttcaag gctatgccct gcaactgcac
     1021 cagttacagg tgctttacaa ggccaagtac ggtccaatgt ggatgtccta cttagggcct
     1081 cagatgcacg tgaacctggc cagtgccccg ctcttggagc aagtgatgcg gcaagagggc
     1141 aagtacccag tacggaacga catggagcta tggaaggagc accgggacca gcacgacctg
     1201 acctatgggc cgttcaccac ggaaggacac cactggtacc agctgcgcca ggctctgaac
     1261 cagcggttgc tgaagccagc ggaagcagcg ctctatacgg atgctttcaa tgaggtgatt
     1321 gatgacttta tgactcgact ggaccagctg cgggcagaga gtgcttcggg gaaccaggtg
     1381 tcggacatgg ctcaactctt ctactacttt gccttggaag ctatttgcta catcctgttc
     1441 gagaaacgca ttggctgcct gcagcgatcc atccccgagg acaccgtgac cttcgtcaga
     1501 tccatcgggt taatgttcca gaactcactc tatgccacct tcctccccaa gtggactcgc
     1561 cccgtgctgc ctttctggaa gcgatacctg gatggttgga atgccatctt ttcctttggg
     1621 aagaagctga ttgatgagaa gctcgaagat atggaggccc aactgcaggc agcagggcca
     1681 gatggcatcc aggtgtctgg ctacctgcac ttcttactgg ccagtggaca gctcagtcct
     1741 cgggaggcca tgggcagcct gcctgagctg ctcatggctg gagtggacac gacatccaac
     1801 acgctgacat gggccctgta ccacctctca aaggaccctg agatccagga ggccttgcac
     1861 gaggaagtgg tgggtgtggt gccagccggg caagtgcccc agcacaagga ctttgcccac
     1921 atgccgttgc tcaaagctgt gcttaaggag actctgcgtc tctaccctgt ggtccccaca
     1981 aactcccgga tcatagaaaa ggaaattgaa gttgatggct tcctcttccc caagaacacc
     2041 cagtttgtgt tctgccacta tgtggtgtcc cgggacccca ctgccttctc tgagcctgaa
     2101 agcttccagc cccaccgctg gctgagaaac agccagcctg ctacccccag gatccagcac
     2161 ccatttggct ctgtgccctt tggctatggg gtccgggcct gcctgggccg caggattgca
     2221 gagctggaga tgcagctact cctcgcaagg ctgatccaga agtacaaggt ggtcctggcc
     2281 ccggagacgg gggagttgaa gagtgtggcc cgcattgtcc tggttcccaa taagaaagtg
     2341 ggcctgcagt tcctgcagag acagtgccac catcaccatc accattagaa ttctgcagat
     2401 atccagcaca gtggcggccg ctcgagtcta gagggcccct taagggttcg atccctaccg
     2461 gttagtaatg agtttgatat ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat
     2521 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     2581 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     2641 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     2701 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     2761 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     2821 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     2881 agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     2941 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     3001 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     3061 gggccgcctc cccgcctgga aacgggggag gctaactgaa acacggaagg agacaatacc
     3121 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt
     3181 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     3241 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     3301 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcagatc
     3361 tgcgcagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     3421 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3481 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3541 atcggggcat ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3601 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3661 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3721 accctatctc ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt
     3781 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     3841 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3901 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3961 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4021 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4081 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     4141 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     4201 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     4261 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4321 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4381 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     4441 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4501 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4561 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4621 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4681 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4741 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4801 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4861 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4921 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4981 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     5041 gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     5101 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5161 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     5221 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5281 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     5341 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5401 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5461 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5521 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5581 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5641 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5701 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5761 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5821 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5881 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5941 cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg
     6001 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6061 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6121 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6181 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     6241 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6301 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6361 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6421 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6481 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6541 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6601 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6661 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6721 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6781 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6841 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6901 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6961 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7021 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7081 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7141 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7201 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7261 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7321 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7381 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7441 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7501 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtc
     7561 gactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
     7621 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
     7681 gaccgacaat tgcatgaaga atctgcttag g
//
