LOCUS       V030614                 9511 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030614
VERSION     V030614
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             706..4170
                     /label="ACLY(NM_001303274)"
                     /note="ACLY(NM_001303274)"
                     /gene="ACLY"
     CDS             4177..4203
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4207..4233
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4237..4263
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    4287..4875
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4941..4989
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5191..5619
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5633..5962
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6029..6823
                     /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    6999..7120
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7169..7185)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7193..7209
                     /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(7217..7247)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7262..7283
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7571..8159)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8330..9190)
                     /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(9191..9295)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatggg ggcggggaaa
      721 agtccggctg ggccgggaca aaagccggat cccgggaagc taccggctgc tggggtgctc
      781 cggattttgc ggggttcgtc gggcctgtgg aagaagcgcc gcgcacggac ttcggcagag
      841 acaggtagag caggtctctc tgcagccatg tcggccaagg caatttcaga gcagacgggc
      901 aaagaactcc tttacaagtt catctgtacc acctcagcca tccagaatcg gttcaagtat
      961 gctcgggtca ctcctgacac agactgggcc cgcttgctgc aggaccaccc ctggctgctc
     1021 agccagaact tggtagtcaa gccagaccag ctgatcaaac gtcgtggaaa acttggtctc
     1081 gttggggtca acctcactct ggatggggtc aagtcctggc tgaagccacg gctgggacag
     1141 gaagccacag ttggcaaggc cacaggcttc ctcaagaact ttctgatcga gcccttcgtc
     1201 ccccacagtc aggctgagga gttctatgtc tgcatctatg ccacccgaga aggggactac
     1261 gtcctgttcc accacgaggg gggtgtggac gtgggtgatg tggacgccaa ggcccagaag
     1321 ctgcttgttg gcgtggatga gaaactgaat cctgaggaca tcaaaaaaca cctgttggtc
     1381 cacgcccctg aagacaagaa agaaattctg gccagtttta tctccggcct cttcaatttc
     1441 tacgaggact tgtacttcac ctacctcgag atcaatcccc ttgtagtgac caaagatgga
     1501 gtctatgtcc ttgacttggc ggccaaggtg gacgccactg ccgactacat ctgcaaagtg
     1561 aagtggggtg acatcgagtt ccctcccccc ttcgggcggg aggcatatcc agaggaagcc
     1621 tacattgcag acctcgatgc caaaagtggg gcaagcctga agctgacctt gctgaacccc
     1681 aaagggagga tctggaccat ggtggccggg ggtggcgcct ctgtcgtgta cagcgatacc
     1741 atctgtgatc tagggggtgt caacgagctg gcaaactatg gggagtactc aggcgccccc
     1801 agcgagcagc agacctatga ctatgccaag actatcctct ccctcatgac ccgagagaag
     1861 cacccagatg gcaagatcct catcattgga ggcagcatcg caaacttcac caacgtggct
     1921 gccacgttca agggcatcgt gagagcaatt cgagattacc agggccccct gaaggagcac
     1981 gaagtcacaa tctttgtccg aagaggtggc cccaactatc aggagggctt acgggtgatg
     2041 ggagaagtcg ggaagaccac tgggatcccc atccatgtct ttggcacaga gactcacatg
     2101 acggccattg tgggcatggc cctgggccac cggcccatcc ccaaccagcc acccacagcg
     2161 gcccacactg caaacttcct cctcaacgcc agcgggagca catcgacgcc agcccccagc
     2221 aggacagcat ctttttctga gtccagggcc gatgaggtgg cgcctgcaaa gaaggccaag
     2281 cctgccatgc cacaagattc agtcccaagt ccaagatccc tgcaaggaaa gagcaccacc
     2341 ctcttcagcc gccacaccaa ggccattgtg tggggcatgc agacccgggc cgtgcaaggc
     2401 atgctggact ttgactatgt ctgctcccga gacgagccct cagtggctgc catggtctac
     2461 cctttcactg gggaccacaa gcagaagttt tactgggggc acaaagagat cctgatccct
     2521 gtcttcaaga acatggctga tgccatgagg aagcatccgg aggtagatgt gctcatcaac
     2581 tttgcctctc tccgctctgc ctatgacagc accatggaga ccatgaacta tgcccagatc
     2641 cggaccatcg ccatcatagc tgaaggcatc cctgaggccc tcacgagaaa gctgatcaag
     2701 aaggcggacc agaagggagt gaccatcatc ggacctgcca ctgttggagg catcaagcct
     2761 gggtgcttta agattggcaa cacaggtggg atgctggaca acatcctggc ctccaaactg
     2821 taccgcccag gcagcgtggc ctatgtctca cgttccggag gcatgtccaa cgagctcaac
     2881 aatatcatct ctcggaccac ggatggcgtc tatgagggcg tggccattgg tggggacagg
     2941 tacccgggct ccacattcat ggatcatgtg ttacgctatc aggacactcc aggagtcaaa
     3001 atgattgtgg ttcttggaga gattgggggc actgaggaat ataagatttg ccggggcatc
     3061 aaggagggcc gcctcactaa gcccatcgtc tgctggtgca tcgggacgtg tgccaccatg
     3121 ttctcctctg aggtccagtt tggccatgct ggagcttgtg ccaaccaggc ttctgaaact
     3181 gcagtagcca agaaccaggc tttgaaggaa gcaggagtgt ttgtgccccg gagctttgat
     3241 gagcttggag agatcatcca gtctgtatac gaagatctcg tggccaatgg agtcattgta
     3301 cctgcccagg aggtgccgcc cccaaccgtg cccatggact actcctgggc cagggagctt
     3361 ggtttgatcc gcaaacctgc ctcgttcatg accagcatct gcgatgagcg aggacaggag
     3421 ctcatctacg cgggcatgcc catcactgag gtcttcaagg aagagatggg cattggcggg
     3481 gtcctcggcc tcctctggtt ccagaaaagg ttgcctaagt actcttgcca gttcattgag
     3541 atgtgtctga tggtgacagc tgatcacggg ccagccgtct ctggagccca caacaccatc
     3601 atttgtgcgc gagctgggaa agacctggtc tccagcctca cctcggggct gctcaccatc
     3661 ggggatcggt ttgggggtgc cttggatgca gcagccaaga tgttcagtaa agcctttgac
     3721 agtggcatta tccccatgga gtttgtgaac aagatgaaga aggaagggaa gctgatcatg
     3781 ggcattggtc accgagtgaa gtcgataaac aacccagaca tgcgagtgca gatcctcaaa
     3841 gattacgtca ggcagcactt ccctgccact cctctgctcg attatgcact ggaagtagag
     3901 aagattacca cctcgaagaa gccaaatctt atcctgaatg tagatggtct catcggagtc
     3961 gcatttgtag acatgcttag aaactgtggg tcctttactc gggaggaagc tgatgaatat
     4021 attgacattg gagccctcaa tggcatcttt gtgctgggaa ggagtatggg gttcattgga
     4081 cactatcttg atcagaagag gctgaagcag gggctgtatc gtcatccgtg ggatgatatt
     4141 tcatatgttc ttccggaaca catgagcatg ctcgagtacc catacgacgt acctgattat
     4201 gcaacttatc cttatgatgt acctgattat gctagctatc cttatgatgt cccagactac
     4261 gcatagtgag ggcccgatat ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat
     4321 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     4381 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     4441 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     4501 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     4561 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     4621 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     4681 agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     4741 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     4801 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     4861 gggccgcctc cccgcctgga aacgggggag gctaactgaa acacggaagg agacaatacc
     4921 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt
     4981 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     5041 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     5101 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcagatc
     5161 tgcgcagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     5221 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     5281 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     5341 atcggggcat ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     5401 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     5461 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     5521 accctatctc ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt
     5581 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     5641 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     5701 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     5761 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     5821 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     5881 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     5941 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     6001 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     6061 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     6121 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     6181 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     6241 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     6301 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     6361 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     6421 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     6481 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     6541 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     6601 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     6661 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     6721 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     6781 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     6841 gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     6901 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     6961 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     7021 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     7081 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     7141 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     7201 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     7261 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     7321 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     7381 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     7441 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     7501 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     7561 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     7621 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     7681 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     7741 cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg
     7801 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     7861 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     7921 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     7981 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     8041 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     8101 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     8161 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     8221 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     8281 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     8341 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     8401 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     8461 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     8521 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     8581 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     8641 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     8701 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     8761 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     8821 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     8881 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     8941 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     9001 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     9061 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     9121 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     9181 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     9241 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     9301 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtc
     9361 gactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
     9421 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
     9481 gaccgacaat tgcatgaaga atctgcttag g
//
