LOCUS       V039917                 9378 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039917
VERSION     V039917
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..4058
                     /label="Acly(NM_016987)"
                     /note="Acly(NM_016987)"
                     /gene="Acly"
     CDS             4065..4130
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4154..4742
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4808..4856
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5058..5486
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5500..5829
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5896..6690
                     /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    6866..6987
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7036..7052)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7060..7076
                     /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(7084..7114)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7129..7150
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7438..8026)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8197..9057)
                     /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(9058..9162)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgtc agccaaggca atttcagagc
      781 agaccggcaa agaactcctt tacaagtaca tctgtaccac ctcagccatc cagaaccggt
      841 tcaagtatgc ccgggttact cccgacacag actgggccca tctcctgcag gaccacccct
      901 ggctgcttag ccagagcttg gtagtcaagc cggaccagct gatcaaacgt cgaggaaagc
      961 ttggtctagt cggggtcaac ctctctctgg atggagtcaa atcctggctg aaacctcgac
     1021 tgggacatga ggccaccgtc ggcaaggcca aaggcttcct caagaacttt ctgattgagc
     1081 ccttcgtccc ccacagtcag gcggaggagt tctacgtgtg catctatgct acccgggaag
     1141 gagactacgt cctgttccac catgaagggg gtgtggatgt gggcgatgtg gacaccaaag
     1201 cccagaagct gcttgtgggt gtggacgaga aactgaacgc tgaagacatt aagagacacc
     1261 tgttggtcca cgcccccgaa gacaagaaag aaatcctggc cagcttcatc tccggcctat
     1321 tcaatttcta cgaagatctt tacttcacct accttgagat caaccccctt gtggtgacca
     1381 aagatggtgt ctacatcctt gacctggcgg ccaaggtgga cgccactgct gactacatct
     1441 gcaaagtcaa gtggggtgat atagagttcc ctcccccctt tgggcgtgag gcatacccag
     1501 aggaagccta cattgcagac ctggatgcca aaagtggggc gagcttgaag ctgaccttgc
     1561 tgaaccccaa ggggcggatc tggaccatgg ttgccggggg tggcgcctct gtcgtgtaca
     1621 gtgataccat ctgtgatctt ggaggtgtca acgaactggc gaattacggg gagtactctg
     1681 gtgcccccag tgaacaacag acctatgact acgccaagac catcctctca cttatgactc
     1741 gagagaagca cccggatggc aagatcctca tcattggagg cagcattgca aacttcacca
     1801 acgtggccgc caccttcaag ggcattgtga gagcaattcg agattaccag ggtcccctga
     1861 aggagcacga ggtcaccatc tttgttcgaa gaggtggccc gaactatcaa gagggattac
     1921 gagtgatggg agaagttggg aagaccactg gaatccccat ccatgtcttt ggcacagaaa
     1981 ctcacatgac ggccattgtg ggcatggccc tgggccaccg gcccattccc aaccagccac
     2041 ccacagcggc tcacactgcc aacttcctcc ttaatgccag tgggagcaca tcgacaccag
     2101 cacccagcag gacagcgtct ttttccgagt ccagagctga cgaggtggcc cctgcaaaga
     2161 aagccaagcc agccatgccc caagattcag tcccaagtcc aagatccctg caaggaaaga
     2221 gtgccaccct cttcagccga cataccaagg ctatcgtgtg gggcatgcag acccgggctg
     2281 tgcaaggcat gctggacttt gactacgtgt gctcccgaga tgagccttca gtggctgcta
     2341 tggtctaccc gttcacgggg gatcataagc agaagtttta ctggggacac aaggaaatcc
     2401 tgatccctgt cttcaagaac atggctgacg ccatgaaaaa gcatccggag gtagacgtgc
     2461 tgatcaactt tgcatctctg cgatcggctt atgacagcac catggagacc atgaactatg
     2521 cacagatccg gaccatagcc atcatagcag aaggcatccc tgaggctctc acacggaagc
     2581 tcatcaagaa ggcagaccag aagggcgtga ccatcattgg gccagccacg gttgggggca
     2641 tcaagcctgg atgctttaag attgggaata ctggtgggat gctggacaac atcctggcct
     2701 ccaaactgta tcgcccaggc agtgtggcct acgtctcgcg ttcaggaggc atgtctaacg
     2761 aactcaataa tatcatctct cggaccacag atggtgtcta cgagggtgtt gccatcggcg
     2821 gggacaggta ccctgggtcc acattcatgg atcacgtgct gcgttaccaa gacactccag
     2881 gagtcaagat gattgtagtt cttggggaga tagggggtac agaagaatat aagatctgcc
     2941 ggggcatcaa ggagggccgc ctcaccaagc cagtggtctg ctggtgcatc gggacctgtg
     3001 ccaccatgtt ctcttctgag gtccagtttg gccacgctgg ggcttgtgcc aaccaggctt
     3061 ctgaaacggc agtagccaag aaccaggcct tgaaggaagc gggagtgttt gtgccccgaa
     3121 gctttgatga gctcggagaa atcattcagt ccgtgtatga agatcttgtg gccaaaggcg
     3181 ccattgtacc tgctcaggaa gtgccacctc caacagtacc catggactac tcttgggcca
     3241 gggagctggg tttaatccga aaacctgcct cattcatgac cagcatctgt gacgagcggg
     3301 ggcaggaact catttatgcg ggcatgccca tcaccgaggt cttcaaggaa gagatgggca
     3361 ttggtggtgt cctgggcctc ctctggttcc agagaaggtt gcccaagtat tcctgccagt
     3421 tcattgagat gtgtctcatg gtcaccgctg atcacgggcc agctgtctcc ggggcccata
     3481 acactatcat ctgtgctcgg gctgggaagg acctggtctc cagcctcacc tcagggctgc
     3541 tcaccattgg ggaccggttt gggggtgcct tggacgcagc agcgaagatg ttcagtaaag
     3601 cctttgacag cggcattatt cccatggagt ttgtgaacaa gatgaagaag gaggggaaac
     3661 tgatcatggg catcggccat cgagtcaaat cgataaacaa cccagacatg cgagtgcaga
     3721 tcctcaaaga ctttgtcaaa cagcacttcc ccgccacccc gctgctcgac tatgcactgg
     3781 aagtggagaa aatcaccacc tcaaagaagc caaatcttat cctgaacgtg gatggtttca
     3841 tcggcgttgc gtttgtggac atgcttagga actgtggctc cttcacccgg gaggaagctg
     3901 acgagtatgt tgacattgga gccctcaatg gcgtctttgt gctgggaagg agtatgggct
     3961 tcatcgggca ctatcttgac cagaagaggc tgaagcaagg gctgtatcgt cacccctggg
     4021 acgacatttc ctatgttctc ccggaacaca tgagcatggg atccgactac aaagaccatg
     4081 acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc
     4141 ccgatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     4201 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     4261 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     4321 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     4381 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     4441 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     4501 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     4561 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     4621 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     4681 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     4741 gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     4801 gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa
     4861 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     4921 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     4981 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg
     5041 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     5101 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     5161 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc
     5221 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     5281 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     5341 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     5401 ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct
     5461 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     5521 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     5581 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     5641 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     5701 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     5761 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     5821 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     5881 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     5941 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     6001 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     6061 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     6121 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     6181 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     6241 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     6301 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     6361 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     6421 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     6481 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     6541 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     6601 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     6661 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga
     6721 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     6781 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     6841 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     6901 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     6961 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     7021 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     7081 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     7141 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     7201 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     7261 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     7321 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     7381 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     7441 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     7501 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     7561 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     7621 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     7681 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     7741 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     7801 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     7861 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     7921 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     7981 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     8041 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     8101 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     8161 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     8221 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     8281 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     8341 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     8401 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     8461 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     8521 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     8581 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     8641 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     8701 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     8761 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     8821 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     8881 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     8941 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     9001 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     9061 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     9121 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     9181 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca
     9241 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     9301 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     9361 atgaagaatc tgcttagg
//
