LOCUS       V041448                 8724 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041448
VERSION     V041448
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        50..1228
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
                     /label="EF-1α promoter"
     CDS             1293..3395
                     /label="ACSS2(NM_018677)"
                     /note="ACSS2(NM_018677)"
                     /gene="ACSS2"
     CDS             3411..3476
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3500..4088
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4154..4202
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4404..4832
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4846..5175
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5242..6036
                     /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    6212..6333
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6382..6398)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6406..6422
                     /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(6430..6460)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6475..6496
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6784..7372)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7543..8403)
                     /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(8404..8508)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg gctccggtgc
       61 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
      121 caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
      181 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
      241 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc
      301 ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg
      361 gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag
      421 gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg
      481 gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc
      541 tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct
      601 tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga
      661 cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc
      721 gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc
      781 gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc
      841 ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc
      901 gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt
      961 cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag
     1021 cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca
     1081 cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga
     1141 atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag
     1201 tttttttctt ccatttcagg tgtcgtgagg aattagcttg gtactggcta gcgtttaaac
     1261 ttaagcttgg taccgagctc ggatccgcca ccatggggct tcctgaggag cgggtccgga
     1321 gcggcagcgg gagccggggc caggaggaag ctggagccgg aggccgggcg cggagttggt
     1381 ctccgccgcc cgaggtcagc cgctccgcgc acgtcccctc gctgcagcgc taccgcgagc
     1441 tgcaccggcg ctccgtggag gagccgcggg aattctgggg agacattgcc aaggaatttt
     1501 actggaagac tccatgccct ggcccattcc ttcggtacaa ctttgatgtg actaaaggga
     1561 aaatcttcat tgagtggatg aaaggagcaa ctaccaacat ctgctacaat gtactggatc
     1621 gaaatgtcca tgagaaaaag cttggagata aagttgcttt ttactgggag ggcaatgagc
     1681 caggggagac cactcagatc acataccatc agcttctggt ccaagtgtgt cagttcagca
     1741 atgttctccg aaaacagggc attcagaagg gggaccgagt ggccatctac atgcctatga
     1801 tcccagagct tgtggtggcc atgctggcat gtgcccgcat tggggctttg cactccattg
     1861 tgtttgcagg cttctcttca gagtctctat gtgaacggat cttggattcc agctgcagtc
     1921 ttctcatcac tacagatgcc ttctacaggg gggaaaagct tgtgaacctg aaggagctgg
     1981 ctgacgaggc cctgcagaag tgtcaggaga agggtttccc agtaagatgc tgcattgtgg
     2041 tcaagcacct ggggcgggca gagctcggca tgggtgactc caccagccag tcccccccaa
     2101 ttaagaggtc atgcccagat gtgcagatct catggaacca agggattgac ttgtggtggc
     2161 atgagctcat gcaagaggca ggggatgagt gtgagcccga gtggtgtgat gccgaggacc
     2221 cactcttcat cctgtacacc agtggctcca caggcaaacc caagggtgtg gttcacacag
     2281 ttgggggcta catgctctat gtagccacaa ccttcaagta tgtgtttgac ttccatgcag
     2341 aggatgtgtt ctggtgcacg gcagacattg gttggatcac tggtcattcc tacgtcacct
     2401 atgggccact ggccaatggt gccaccagtg ttttgtttga ggggattccc acatatccgg
     2461 acgtgaaccg cctgtggagc attgtggaca aatacaaggt gaccaagttc tacacagcac
     2521 ccacagccat ccgtctgctc atgaagtttg gagatgagcc tgtcaccaag catagccggg
     2581 catccttgca ggtgttaggc acagtgggtg aacccatcaa ccctgaggcc tggctatggt
     2641 accaccgggt ggtaggtgcc cagcgctgcc ccatcgtgga caccttctgg caaacagaga
     2701 caggtggcca catgttgact ccccttcctg gtgccacacc catgaaaccc ggttctgcta
     2761 ctttcccatt ctttggtgta gctcctgcaa tcctgaatga gtccggggaa gagttggaag
     2821 gtgaagctga aggttatctg gtgttcaagc agccctggcc agggatcatg cgcacagtct
     2881 atgggaacca cgaacgcttt gagacaacct actttaagaa gtttcctgga tactatgtta
     2941 caggagatgg ctgccagcgg gaccaggatg gctattactg gatcactggc aggattgatg
     3001 acatgctcaa tgtatctgga cacctgctga gtacagcaga ggtggagtca gcacttgtgg
     3061 aacatgaggc tgttgcagag gcagctgtgg tgggccaccc tcatcctgtg aagggtgaat
     3121 gcctctactg ctttgtcacc ttgtgtgatg gccacacctt cagccccaag ctcaccgagg
     3181 agctcaagaa gcagattaga gaaaagattg gccccattgc cacaccagac tacatccaga
     3241 atgcacctgg cttgcctaaa acccgctcag ggaaaatcat gaggcgagtg cttcggaaga
     3301 ttgctcagaa tgaccatgac ctcggggaca tgtctactgt ggctgaccca tctgtcatca
     3361 gtcacctctt cagccaccgc tgcctgacca tccagctgcg gccgctcgag gactacaaag
     3421 accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt
     3481 gagggcccga tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     3541 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     3601 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     3661 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     3721 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     3781 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     3841 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     3901 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     3961 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     4021 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     4081 ctccccgcct ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga
     4141 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt
     4201 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     4261 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     4321 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag
     4381 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4441 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4501 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4561 catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4621 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4681 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4741 ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa
     4801 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     4861 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4921 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4981 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     5041 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     5101 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     5161 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     5221 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     5281 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     5341 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     5401 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     5461 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     5521 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5581 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5641 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     5701 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     5761 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5821 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5881 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5941 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     6001 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat
     6061 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     6121 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     6181 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     6241 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     6301 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     6361 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6421 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6481 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6541 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6601 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6661 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6721 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6781 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6841 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6901 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6961 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     7021 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     7081 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7141 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7201 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     7261 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7321 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7381 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7441 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7501 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7561 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7621 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7681 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7741 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7801 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7861 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7921 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7981 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     8041 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     8101 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     8161 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     8221 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     8281 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8341 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8401 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8461 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8521 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc
     8581 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     8641 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     8701 aattgcatga agaatctgct tagg
//
