LOCUS       V042130                 8047 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042130
VERSION     V042130
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             964..3537
                     /label="Eef2(NM_007907)"
                     /note="Eef2(NM_007907)"
                     /gene="Eef2"
     CDS             3544..3609
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3647..3871
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3917..4345
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4359..4688
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4755..5549
                     /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    5723..5844
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5893..5909)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5917..5933
                     /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(5941..5971)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5986..6007
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6295..6880)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7051..7911)
                     /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(7912..8016)
                     /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 atccactagt ccagtgtggt ggaattcgcc
      961 accatggtga acttcacagt agatcagatc cgtgccatca tggacaagaa agccaacatc
     1021 cggaacatgt cagtcatcgc ccatgtggac cacggcaagt ccacgctgac cgactccctt
     1081 gtgtgcaagg ctggcatcat tgcctctgcc cgagctgggg agacgcgctt cactgacact
     1141 cgcaaggatg agcaggagcg ctgcatcaca atcaaatcca ccgccatctc cctcttctac
     1201 gagctctctg agaacgacct gaacttcatt aagcagagca aggatggctc gggcttcctc
     1261 atcaacctca tcgactctcc aggccatgtg gacttctctt cagaggtgac agctgccttg
     1321 cgtgtcaccg atggagctct ggtggtggtg gactgtgtgt ctggcgtgtg tgtgcagaca
     1381 gaaactgtgc tgcgccaggc catcgctgaa cgcatcaagc ccgtcctgat gatgaacaag
     1441 atggaccggg ccctgctgga gctgcagctg gagcccgagg agctctacca gaccttccag
     1501 cgcatcgtgg agaacgtcaa cgtcatcatc tctacctacg gcgagggcga gagtgggccc
     1561 atgggcaata tcatgattga ccccgtcctg ggcaccgtag gctttggttc tggcctgcat
     1621 ggctgggcct tcaccctgaa gcagtttgcg gagatgtatg tggccaagtt tgcagccaag
     1681 ggtgagggcc agctgagcgc agccgagcgt gccaagaaag tagaggacat gatgaagaag
     1741 ctgtggggag accggtactt tgatccggcc aatggcaagt tcagtaagtc agccaatagc
     1801 ccagatggga aaaaacttcc ccgcaccttc tgccagctca tcctggaccc catcttcaag
     1861 gtgttcgacg ccatcatgaa cttcaggaag gaggagacag ccaagctgat cgagaagctg
     1921 gacatcaagc tggacagcga ggacaaagac aaggagggca agccattgct caaggctgtg
     1981 atgcgccgct ggctgcctgc aggggacgcc ctcctgcaga tgatcaccat ccacttacca
     2041 tcccccgtca ctgcacagaa gtaccgttgt gagctgctat acgaggggcc tcccgacgat
     2101 gaggccgcca tgggtattaa gagctgcgac cccaaaggcc cacttatgat gtacatttcc
     2161 aagatggtgc caacctctga caaaggccgc ttctatgcct ttggtagagt gttctctggg
     2221 gtggtgtcca cgggcctgaa ggtccggatc atgggcccca actacacgcc tgggaagaaa
     2281 gaggacctat acctgaagcc tatccagaga accattctga tgatgggccg ctacgtggag
     2341 ccgattgagg acgtgccatg tggaaacatt gtggggctgg tcggagtgga ccagttcctt
     2401 gtgaagacgg ggaccatcac tacctttgag cacgctcaca acatgcgcgt gatgaagttc
     2461 agcgtcagcc ctgtcgtccg tgtggcagtg gaggccaaga acccagctga cctgcccaag
     2521 ctggtggagg ggctgaagcg gctggctaag tctgacccta tggtgcagtg catcattgag
     2581 gagtctggag aacatattat tgctggcgcc ggtgagctgc acctggagat ctgccttaag
     2641 gacctggagg aggaccatgc ctgcatcccc atcaagaaat ctgaccctgt tgtgtcatat
     2701 cgggagacag tcagtgagga atccaacgtg ctgtgtctgt ccaagtcccc caataagcac
     2761 aaccggctgt acatgaaggc caggcccttc cctgatggcc tggcagagga catcgataag
     2821 ggtgaggtgt ctgcccgcca ggagctcaag gcacgtgccc gctacctggc cgaaaagtat
     2881 gagtgggacg ttgctgaagc ccgcaagatc tggtgctttg gccctgatgg cactggcccc
     2941 aacattctca ccgacatcac caagggtgtg cagtacctga atgagatcaa ggacagtgtg
     3001 gtggctggct tccagtgggc tactaaggag ggcgctctct gtgaggaaaa catgcgtggt
     3061 gtgcggtttg atgttcatga tgtgaccctg catgctgatg ccattcaccg gggaggtggc
     3121 cagatcatcc ccacagcacg ccgctgcctg tatgccagtg tgctgaccgc acagccccgc
     3181 ctcatggagc ctatctatct ggtggagatc cagtgtcctg agcaagtggt gggtggcatc
     3241 tacggtgtcc tgaacaggaa gcgtggccat gtgtttgagg agtcccaggt ggctggtacc
     3301 cccatgtttg tggtcaaggc atacctgcct gtcaatgagt cctttggctt caccgctgat
     3361 ctgcgatcca acaccggcgg ccaggccttc ccccagtgcg tgtttgacca ctggcagatc
     3421 ctgcctgggg atccttttga caacagcagc cgccccagcc aagtggtagc tgagacgcgc
     3481 aagcgcaagg gcctgaaaga gggcatccca gcgctggaca acttcctgga caaactgctc
     3541 gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac
     3601 gatgacaagt agtgagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     3661 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3721 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     3781 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     3841 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     3901 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3961 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4021 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4081 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4141 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4201 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4261 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4321 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     4381 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4441 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4501 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4561 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4621 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4681 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     4741 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4801 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4861 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4921 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4981 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     5041 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     5101 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     5161 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     5221 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     5281 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     5341 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5401 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5461 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5521 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     5581 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5641 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5701 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5761 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5821 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5881 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5941 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     6001 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     6061 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     6121 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     6181 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6241 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6301 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6361 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6421 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6481 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6541 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6601 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6661 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6721 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6781 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt
     6841 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6901 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6961 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7021 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7081 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7141 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7201 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7261 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7321 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7381 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7441 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7501 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7561 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7621 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7681 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7741 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7801 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7861 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7921 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7981 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8041 tgacgtc
//
