LOCUS       V018621                 7900 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018621
VERSION     V018621
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             997..3420
                     /label="LONP2(NM_001300948)"
                     /note="LONP2(NM_001300948)"
                     /gene="LONP2"
     polyA_signal    3500..3724
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3770..4198
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4212..4541
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4608..5402
                     /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    5576..5697
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5746..5762)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5770..5786
                     /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(5794..5824)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5839..5860
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6148..6733)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6904..7764)
                     /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(7765..7869)
                     /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 ggtaccatgt catcagtgag ccccatccag
     1021 atccccagtc gcctcccgct gctgctcacc cacgagggcg tcctgctgcc cggctccacc
     1081 atgcgcacca gcgtggactc ggcccgcaac ctgcagctgg tgcggagccg ccttctgaag
     1141 ggcacgtcgc tgcaaagcac catcctgggc gtcatcccca acacgcctga ccccgccagc
     1201 gacgcgcagg acctgccgcc gctgcacagg attggcacag ctgcactggc cgttcaggtt
     1261 gtgggcagta actggcccaa gccccactac actctgttga ttacaggcct atgccgtttc
     1321 cagattgtac aggtcttaaa agagaagcca tatcccattg ctgaagtgga gcagttggac
     1381 cgacttgagg agtttcccaa cacctgtaaa atgagggagg agctaggaga actatcagag
     1441 cagttttaca aatatgcagt acaaatttta gatgctgtga gcctagagga gcggttcaag
     1501 atgactatac cactgcttgt cagacaaatt gaaggcctga aattgcttca aaaaaccaga
     1561 aaacccaagc aagatgatga taagagggtt atagcaatac gccctattag gagaattaca
     1621 catatctcag gtactttaga agatgaagat gaagatgaag ataatgatga cattgtcatg
     1681 ctagagaaaa aaatacgaac atctagtatg ccagagcagg cccataaagt ctgtgtcaaa
     1741 gagataaaga gactcaaaaa aatgcctcag tcaatgccag aatatgctct gactagaaat
     1801 tatttggaac ttatggtaga acttccttgg aacaaaagta caactgaccg cctggacatt
     1861 agggcagccc ggattcttct ggataatgac cattacgcca tggaaaaatt gaagaaaaga
     1921 gtactggaat acttggctgt cagacagctc aaaaataacc tgaagggccc aatcctatgc
     1981 tttgttggcc ctcctggagt tggtaaaaca agtgtgggaa gatcagtggc caagactcta
     2041 ggtcgagagt tccacaggat tgcacttgga ggagtatgtg atcagtctga cattcgagga
     2101 cacaggcgca cctatgttgg cagcatgcct ggtcgcatca tcaacggctt gaagactgtg
     2161 ggagtgaaca acccagtgtt cctattagat gaggttgaca aactgggaaa aagtctacag
     2221 ggtgatccag cagcagctct gcttgaggtg ttggatcctg aacaaaacca taacttcaca
     2281 gatcattatc taaatgtggc ctttgacctt tctcaagttc tttttatagc tactgccaac
     2341 accactgcta ccattccagc tgccttgttg gacagaatgg agatcattca ggttccaggt
     2401 tatacacagg aggagaagat agagattgcc cataggcact tgatccccaa gcagctggaa
     2461 caacatgggc tgactccaca gcagattcag ataccccagg tcaccactct tgacatcatc
     2521 accaggtata ccagagaggc aggggttcgt tctctggata gaaaacttgg ggccatttgc
     2581 cgagctgtgg ccgtgaaggt ggcagaagga cagcataagg aagccaagtt ggaccgttct
     2641 gatgtgactg agagagaagg ttgcagagaa cacatcttag aagatgaaaa acctgaatct
     2701 atcagtgaca ctactgactt ggctctacca cctgaaatgc cgattttgat tgatttccat
     2761 gctctgaaag acatccttgg gcccccgatg tatgaaatgg aggtatctca gcgtttgagt
     2821 cagccaggag tagcaatagg tttggcttgg actcccttag gtggagaaat catgttcgtg
     2881 gaggcgagtc gaatggatgg cgagggccag ttaactctga ccggccagct cggggacgtg
     2941 atgaaggagt ccgcccacct cgctatcagc tggctccgca gcaacgcaaa gaagtaccag
     3001 ctgaccaatg cttttggaag ttttgatctt cttgacaaca cagacatcca tctgcacttc
     3061 ccagctggag ctgtcacaaa agatggacca tctgctggag ttaccatagt aacctgtctc
     3121 gcctcacttt ttagtgggcg gctggtacgt tcagatgtag ccatgactgg agaaattaca
     3181 ctgagaggtc ttgttcttcc agtgggtgga attaaagaca aagtgctggc ggcacacaga
     3241 gcgggactga agcaagtcat tattcctcgg agaaatgaaa aagaccttga gggaatccca
     3301 ggcaacgtac gacaggattt aagttttgtc acagcaagct gcctggatga ggttcttaat
     3361 gcagcttttg atggtggctt tactgtcaag accagacctg gtctgttaaa tagcaaactg
     3421 taggaattct gcagatatcc agcacagtgg cggccgctcg agtctagagg gcccgtttaa
     3481 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     3541 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     3601 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     3661 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     3721 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     3781 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3841 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3901 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3961 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4021 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4081 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     4141 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     4201 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4261 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     4321 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     4381 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4441 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     4501 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     4561 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     4621 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4681 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4741 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     4801 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4861 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4921 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4981 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     5041 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     5101 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     5161 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     5221 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     5281 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     5341 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     5401 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5461 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5521 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     5581 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5641 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5701 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     5761 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5821 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5881 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5941 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6001 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6061 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6121 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6181 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6241 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6301 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6361 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6421 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6481 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6541 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     6601 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6661 caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6721 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6781 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6841 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6901 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6961 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7021 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7081 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7141 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7201 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7261 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7321 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7381 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7441 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7501 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7561 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7621 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7681 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7741 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7801 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7861 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
