LOCUS       V026688                 8747 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026688
VERSION     V026688
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"
     CDS             944..970
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             977..4279
                     /label="ACLY(NM_001096)"
                     /note="ACLY(NM_001096)"
                     /gene="ACLY"
     CDS             4286..4309
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     polyA_signal    4347..4571
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4617..5045
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5059..5388
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5455..6249
                     /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    6423..6544
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6593..6609)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6617..6633
                     /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(6641..6671)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6686..6707
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6995..7580)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7751..8611)
                     /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(8612..8716)
                     /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 aagcttggta ccgagctcgg atccgccacc atgtacccat acgatgttcc
      961 agattacgct gaattcatgt cggccaaggc aatttcagag cagacgggca aagaactcct
     1021 ttacaagttc atctgtacca cctcagccat ccagaatcgg ttcaagtatg ctcgggtcac
     1081 tcctgacaca gactgggccc gcttgctgca ggaccacccc tggctgctca gccagaactt
     1141 ggtagtcaag ccagaccagc tgatcaaacg tcgtggaaaa cttggtctcg ttggggtcaa
     1201 cctcactctg gatggggtca agtcctggct gaagccacgg ctgggacagg aagccacagt
     1261 tggcaaggcc acaggcttcc tcaagaactt tctgatcgag cccttcgtcc cccacagtca
     1321 ggctgaggag ttctatgtct gcatctatgc cacccgagaa ggggactacg tcctgttcca
     1381 ccacgagggg ggtgtggacg tgggtgatgt ggacgccaag gcccagaagc tgcttgttgg
     1441 cgtggatgag aaactgaatc ctgaggacat caaaaaacac ctgttggtcc acgcccctga
     1501 agacaagaaa gaaattctgg ccagttttat ctccggcctc ttcaatttct acgaggactt
     1561 gtacttcacc tacctcgaga tcaatcccct tgtagtgacc aaagatggag tctatgtcct
     1621 tgacttggcg gccaaggtgg acgccactgc cgactacatc tgcaaagtga agtggggtga
     1681 catcgagttc cctcccccct tcgggcggga ggcatatcca gaggaagcct acattgcaga
     1741 cctcgatgcc aaaagtgggg caagcctgaa gctgaccttg ctgaacccca aagggaggat
     1801 ctggaccatg gtggccgggg gtggcgcctc tgtcgtgtac agcgatacca tctgtgatct
     1861 agggggtgtc aacgagctgg caaactatgg ggagtactca ggcgccccca gcgagcagca
     1921 gacctatgac tatgccaaga ctatcctctc cctcatgacc cgagagaagc acccagatgg
     1981 caagatcctc atcattggag gcagcatcgc aaacttcacc aacgtggctg ccacgttcaa
     2041 gggcatcgtg agagcaattc gagattacca gggccccctg aaggagcacg aagtcacaat
     2101 ctttgtccga agaggtggcc ccaactatca ggagggctta cgggtgatgg gagaagtcgg
     2161 gaagaccact gggatcccca tccatgtctt tggcacagag actcacatga cggccattgt
     2221 gggcatggcc ctgggccacc ggcccatccc caaccagcca cccacagcgg cccacactgc
     2281 aaacttcctc ctcaacgcca gcgggagcac atcgacgcca gcccccagca ggacagcatc
     2341 tttttctgag tccagggccg atgaggtggc gcctgcaaag aaggccaagc ctgccatgcc
     2401 acaagattca gtcccaagtc caagatccct gcaaggaaag agcaccaccc tcttcagccg
     2461 ccacaccaag gccattgtgt ggggcatgca gacccgggcc gtgcaaggca tgctggactt
     2521 tgactatgtc tgctcccgag acgagccctc agtggctgcc atggtctacc ctttcactgg
     2581 ggaccacaag cagaagtttt actgggggca caaagagatc ctgatccctg tcttcaagaa
     2641 catggctgat gccatgagga agcatccgga ggtagatgtg ctcatcaact ttgcctctct
     2701 ccgctctgcc tatgacagca ccatggagac catgaactat gcccagatcc ggaccatcgc
     2761 catcatagct gaaggcatcc ctgaggccct cacgagaaag ctgatcaaga aggcggacca
     2821 gaagggagtg accatcatcg gacctgccac tgttggaggc atcaagcctg ggtgctttaa
     2881 gattggcaac acaggtggga tgctggacaa catcctggcc tccaaactgt accgcccagg
     2941 cagcgtggcc tatgtctcac gttccggagg catgtccaac gagctcaaca atatcatctc
     3001 tcggaccacg gatggcgtct atgagggcgt ggccattggt ggggacaggt acccgggctc
     3061 cacattcatg gatcatgtgt tacgctatca ggacactcca ggagtcaaaa tgattgtggt
     3121 tcttggagag attgggggca ctgaggaata taagatttgc cggggcatca aggagggccg
     3181 cctcactaag cccatcgtct gctggtgcat cgggacgtgt gccaccatgt tctcctctga
     3241 ggtccagttt ggccatgctg gagcttgtgc caaccaggct tctgaaactg cagtagccaa
     3301 gaaccaggct ttgaaggaag caggagtgtt tgtgccccgg agctttgatg agcttggaga
     3361 gatcatccag tctgtatacg aagatctcgt ggccaatgga gtcattgtac ctgcccagga
     3421 ggtgccgccc ccaaccgtgc ccatggacta ctcctgggcc agggagcttg gtttgatccg
     3481 caaacctgcc tcgttcatga ccagcatctg cgatgagcga ggacaggagc tcatctacgc
     3541 gggcatgccc atcactgagg tcttcaagga agagatgggc attggcgggg tcctcggcct
     3601 cctctggttc cagaaaaggt tgcctaagta ctcttgccag ttcattgaga tgtgtctgat
     3661 ggtgacagct gatcacgggc cagccgtctc tggagcccac aacaccatca tttgtgcgcg
     3721 agctgggaaa gacctggtct ccagcctcac ctcggggctg ctcaccatcg gggatcggtt
     3781 tgggggtgcc ttggatgcag cagccaagat gttcagtaaa gcctttgaca gtggcattat
     3841 ccccatggag tttgtgaaca agatgaagaa ggaagggaag ctgatcatgg gcattggtca
     3901 ccgagtgaag tcgataaaca acccagacat gcgagtgcag atcctcaaag attacgtcag
     3961 gcagcacttc cctgccactc ctctgctcga ttatgcactg gaagtagaga agattaccac
     4021 ctcgaagaag ccaaatctta tcctgaatgt agatggtctc atcggagtcg catttgtaga
     4081 catgcttaga aactgtgggt cctttactcg ggaggaagct gatgaatata ttgacattgg
     4141 agccctcaat ggcatctttg tgctgggaag gagtatgggg ttcattggac actatcttga
     4201 tcagaagagg ctgaagcagg ggctgtatcg tcatccgtgg gatgatattt catatgttct
     4261 tccggaacac atgagcatgc tcgaggatta caaggatgac gacgataagt agtgagggcc
     4321 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     4381 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     4441 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     4501 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     4561 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     4621 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     4681 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     4741 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     4801 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     4861 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     4921 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     4981 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     5041 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     5101 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     5161 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     5221 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     5281 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     5341 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     5401 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     5461 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     5521 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     5581 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     5641 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     5701 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     5761 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     5821 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     5881 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     5941 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     6001 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     6061 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     6121 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     6181 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     6241 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     6301 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     6361 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     6421 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     6481 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     6541 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     6601 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     6661 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     6721 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     6781 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     6841 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     6901 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     6961 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     7021 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     7081 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     7141 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     7201 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     7261 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     7321 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     7381 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     7441 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     7501 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc
     7561 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     7621 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7681 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7741 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7801 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7861 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7921 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     7981 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     8041 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     8101 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     8161 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     8221 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     8281 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     8341 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     8401 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     8461 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     8521 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     8581 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     8641 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8701 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
