LOCUS       V041865                 8364 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041865
VERSION     V041865
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..3044
                     /label="Hadha(NM_130826)"
                     /note="Hadha(NM_130826)"
                     /gene="Hadha"
     CDS             3051..3116
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3140..3728
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3794..3842
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4044..4472
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4486..4815
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4882..5676
                     /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    5852..5973
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6022..6038)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6046..6062
                     /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(6070..6100)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6115..6136
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6424..7012)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7183..8043)
                     /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(8044..8148)
                     /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 ccaccatggt ggcgtcccgg gcaattggca
      781 gtctcagtcg cttctctgcc ttcaggatcc tgcgctccag aggctgcatt tgccacagct
      841 ttacaacttc ttctgctttg ctgtctagaa cccatattaa ttatggagtc aaaggggatg
      901 tggcagttat tcggattaac tcgcccaatt caaaggtaaa tacattgaat aaagaagtac
      961 aatcagagtt cgtagaagta atgaacgaaa tctgggccaa cgaccaaatc aggagcgccg
     1021 tccttatttc gtcaaagcct ggctgctttg ttgcaggtgc tgacatcaac atgctggcct
     1081 cttgtacaac gccccaagaa gcagcacgaa tatcacaaga aggacagaaa atgtttgaga
     1141 aacttgaaaa gtcaccaaag cctgttgttg ccgccatcag tggatcctgc ttgggaggcg
     1201 gacttgagct tgccatagca tgtcaataca gaatagcaac aaaagacaga aaaacagtat
     1261 taggtgtccc tgaagtgttg ctgggaatct taccaggagc cggaggtacc cagaggctgc
     1321 ccaaaatggt gggtgtgcct gctgcttttg acatgatgct gactggtagg aacattcgtg
     1381 cagacagagc aaagaaaatg ggactggttg accagttggt ggacccgcta ggaccaggaa
     1441 taaaatctcc agaggaaagg acaattgaat acctagaaga agttgcagtt aattttgcca
     1501 aaggcctggc tgacaggaag gtctctgcaa agcagagcaa aggcctgatg gaaaagctga
     1561 catcgtatgc catgactatc ccatttgtca gacaacaggt ttacaaaaca gtggaagaaa
     1621 aagtgaagaa gcagaccaaa ggcctttacc ctgcaccttt gaagataatt gacgctgtga
     1681 agactggact tgagcaagga aatgatgctg gctatcttgc cgaatcagag aaatttggag
     1741 agcttgcatt gaccaaagaa tcaaaagccc tgatggggct ttataatggc caggtcctgt
     1801 gcaagaaaaa taaatttgga gcgccacaga agactgttca gcagctagcc atccttggcg
     1861 cagggctgat gggggctggc attgcccagg tctctgtgga caagggactg aaaactcttc
     1921 ttaaagacac tacagtgaca gggctgggcc ggggacagca acaagtgttc aaaggactga
     1981 atgacaaggt aaagaagaag gcactcacat ccttcgaaag ggactccatc ttcagcaacc
     2041 tgatcgggca gctcgactac aagggcttcg agaaggctga catggtgatt gaggctgtct
     2101 tcgaggacct cgctgttaag cacaaagtgt taaaggaagt ggaaagcgtg actccagaac
     2161 actgtatctt cgccagcaac acatctgctc tcccaatcaa tcaaattgct gctgtgagcc
     2221 aaaggcctga gaaggtgatc ggcatgcact acttctctcc tgtggacaag atgcagcttc
     2281 tagagatcat cacaactgac aaaacctcca aggacaccac agcgtctgcc gtggccgtgg
     2341 gtctcaagca ggggaaggtc atcattgtgg tcaaggacgg acctggcttc tacaccacca
     2401 ggtgtcttgc tcccatgatg tcagaagtca taagaatcct ccaggaagga gttgacccta
     2461 agaagctgga cgccttgacc acaggcttcg gcttccctgt gggtgctgcc accctggcag
     2521 atgaagtagg gatagatgta gcacagcacg tagcagaaga tctaggcaaa gccttcgggg
     2581 agcggtttgg aggtggcagc gtagaactgc tgaaactgat ggtctccaag ggcttcttgg
     2641 gtcgcaagtc tgggaagggc ttctacatct atcagtcggg ctcaaagaat aagaatttga
     2701 attctgaaat agataatatc ttggtaaacc tgaggctgcc tgccaagccc gaggtctcct
     2761 ctgatgaaga catccagtac cgtgtgataa caaggtttgt gaatgaggca gtcctgtgcc
     2821 tacaggaagg gatcctagcc acgcctgaag agggagacat cggagcagtc tttgggcttg
     2881 gctttccccc ttgtctcgga gggcccttcc gctttgtgga tctgtatggt gctcagaagg
     2941 tagtggaccg gctccggaag tatgagtctg cctatgggac acagtttacc ccgtgtcagc
     3001 tactccgcga ccttgctaac aactctagca agaagttcta ccagggatcc gactacaaag
     3061 accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt
     3121 gagggcccga tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     3181 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     3241 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     3301 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     3361 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     3421 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     3481 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     3541 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     3601 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     3661 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     3721 ctccccgcct ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga
     3781 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt
     3841 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     3901 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     3961 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag
     4021 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4081 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4141 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4201 catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4261 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4321 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4381 ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa
     4441 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     4501 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4561 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4621 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4681 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4741 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4801 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     4861 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4921 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4981 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     5041 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     5101 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     5161 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5221 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5281 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     5341 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     5401 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5461 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5521 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5581 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5641 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat
     5701 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5761 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5821 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     5881 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5941 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     6001 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6061 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6121 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6181 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6241 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6301 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6361 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6421 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6481 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6541 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6601 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6661 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6721 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6781 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6841 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     6901 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6961 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7021 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7081 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7141 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7201 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7261 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7321 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7381 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7441 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7501 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7561 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7621 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7681 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7741 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7801 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7861 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7921 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7981 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8041 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8101 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8161 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc
     8221 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     8281 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     8341 aattgcatga agaatctgct tagg
//
