LOCUS       V023555                 7756 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023555
VERSION     V023555
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             1009..3276
                     /label="CPT1A(NM_001031847)"
                     /note="CPT1A(NM_001031847)"
                     /gene="CPT1A"
     polyA_signal    3356..3580
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3626..4054
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4068..4397
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4464..5258
                     /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    5432..5553
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5602..5618)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5626..5642
                     /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(5650..5680)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5695..5716
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6004..6589)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6760..7620)
                     /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(7621..7725)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt ggtaccgagc tcggatccat ggcagaagct
     1021 caccaagctg tggcctttca gttcacggtc actccggacg ggattgacct gcggctgagc
     1081 catgaagctc ttagacaaat ctatctctct ggacttcatt cctggaaaaa gaagttcatc
     1141 agattcaaga acggcatcat cactggcgtg tacccggcaa gcccctccag ttggcttatc
     1201 gtggtggtgg gcgtgatgac aacgatgtac gccaagatcg acccctcgtt aggaataatt
     1261 gcaaaaatca atcggactct ggaaacggcc aactgcatgt ccagccagac gaagaacgtg
     1321 gtcagcggcg tgctgtttgg caccggcctg tgggtggccc tcatcgtcac catgcgctac
     1381 tccctgaaag tgctgctctc ctaccacggg tggatgttca ctgagcacgg caagatgagt
     1441 cgtgccacca agatctggat gggtatggtc aagatctttt caggccgaaa acccatgttg
     1501 tacagcttcc agacatcgct gcctcgcctg ccggtcccgg ctgtcaaaga cactgtgaac
     1561 aggtatctac agtcggtgag gcctcttatg aaggaagaag acttcaaacg gatgacagca
     1621 cttgctcaag attttgctgt cggtcttgga ccaagattac agtggtattt gaagttaaaa
     1681 tcctggtggg ctacaaatta cgtgagcgac tggtgggagg agtacatcta cctccgagga
     1741 cgagggccgc tcatggtgaa cagcaactat tatgccatgg atctgctgta tatccttcca
     1801 actcacattc aggcagcaag agccggcaac gccatccatg ccatcctgct ttacaggcgc
     1861 aaactggacc gggaggaaat caaaccaatt cgtcttttgg gatccacgat tccactctgc
     1921 tccgctcagt gggagcggat gtttaatact tcccggatcc caggagagga gacagacacc
     1981 atccagcaca tgagagacag caagcacatc gtcgtgtacc atcgaggacg ctacttcaag
     2041 gtctggctct accatgatgg gcggctgctg aagccccggg agatggagca gcagatgcag
     2101 aggatcctgg acaatacctc ggagcctcag cccggggagg ccaggctggc agccctcacc
     2161 gcaggagaca gagttccctg ggccaggtgt cgtcaggcct attttggacg tgggaaaaat
     2221 aagcagtctc ttgatgctgt ggagaaagca gcgttcttcg tgacgttaga tgaaactgaa
     2281 gaaggataca gaagtgaaga cccggatacg tcaatggaca gctacgccaa atctctacta
     2341 cacggccgat gttacgacag gtggtttgac aagtcgttca cgtttgttgt cttcaaaaac
     2401 gggaagatgg gcctcaacgc tgaacactcc tgggcagatg cgccgatcgt ggcccacctt
     2461 tgggagtacg tcatgtccat tgacagcctc cagctgggct atgcggagga tgggcactgc
     2521 aaaggcgaca tcaatccgaa cattccgtac cccaccaggc tgcagtggga catcccgggg
     2581 gaatgtcaag aggttataga gacctccctg aacaccgcaa atcttctggc aaacgacgtg
     2641 gatttccatt ccttcccatt cgtagccttt ggtaaaggaa tcatcaagaa atgtcgcacg
     2701 agcccagacg cctttgtgca gctggccctc cagctggcgc actacaagga catgggcaag
     2761 ttttgcctca catacgaggc ctccatgacc cggctcttcc gagaggggag gacggagacc
     2821 gtgcgctcct gcaccactga gtcatgcgac ttcgtgcggg ccatggtgga cccggcccag
     2881 acggtggaac agaggctgaa gttgttcaag ttggcgtctg agaagcatca gcatatgtat
     2941 cgcctcgcca tgaccggctc tgggatcgat cgtcacctct tctgccttta cgtggtgtct
     3001 aaatatctcg ctgtggagtc ccctttcctt aaggaagttt tatctgagcc ttggagatta
     3061 tcaacaagcc agacccctca gcagcaagtg gagctgtttg acttggagaa taacccagag
     3121 tacgtgtcca gcggaggggg ctttggaccg gttgctgatg acggctatgg tgtgtcgtac
     3181 atccttgtgg gagagaacct catcaatttc cacatttctt ccaagttctc ttgccctgag
     3241 acggggatta taagtcaagg accaagttca gatacttgag aattctgcag atatccagca
     3301 cagtggcggc cgctcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg
     3361 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     3421 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     3481 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     3541 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     3601 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3661 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3721 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3781 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3841 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3901 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3961 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     4021 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     4081 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4141 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4201 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4261 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4321 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4381 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     4441 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4501 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4561 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4621 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     4681 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4741 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4801 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4861 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4921 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4981 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     5041 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5101 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     5161 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5221 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     5281 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5341 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5401 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     5461 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5521 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     5581 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5641 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5701 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5761 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5821 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5881 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5941 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6001 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6061 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6121 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6181 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6241 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6301 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6361 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6421 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6481 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6541 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6601 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6661 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6721 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6781 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6841 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6901 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6961 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     7021 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     7081 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     7141 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     7201 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     7261 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     7321 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     7381 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     7441 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7501 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7561 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7621 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7681 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7741 agtgccacct gacgtc
//
