LOCUS       V043890                 7354 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043890
VERSION     V043890
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..2844
                     /label="DTNB(NM_021907)"
                     /note="DTNB(NM_021907)"
                     /gene="DTNB"
     CDS             2851..2916
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2954..3178
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3224..3652
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3666..3995
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4062..4856
                     /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    5030..5151
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5200..5216)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5224..5240
                     /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(5248..5278)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5293..5314
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5602..6187)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6358..7218)
                     /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(7219..7323)
                     /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 accatgattg aggaaagtgg gaacaagcgg aagaccatgg cagagaagag gcagctgttc
     1021 atagaaatgc gtgctcagaa ttttgatgtc atacgactat caacttacag aacagcctgc
     1081 aaattacgat ttgtacaaaa acgatgcaac cttcatcttg ttgatatctg gaacatgatt
     1141 gaagccttcc gagacaatgg ccttaataca ctggaccata ccaccgagat cagtgtgtcc
     1201 cgcctcgaaa ctgtcatctc ctccatctac tatcagttga acaagcgcct tccttctact
     1261 caccaaatta gtgtggaaca atctatcagc ctcctcctca actttatgat tgctgcatat
     1321 gacagtgagg gccgaggcaa gttgacggta ttttcagtta aagctatgtt agcaaccatg
     1381 tgtggtggaa aaatgctgga caaattgaga tatgttttct cccagatgtc agattccaat
     1441 ggcttaatga tatttagcaa gtttgaccag tttctgaagg aagttctgaa gctcccaaca
     1501 gctgtctttg aagggccatc ttttggttac acagagcact cagtccgcac ctgttttcca
     1561 cagcagagaa agataatgct aaatatgttt ttagacacaa tgatggctga ccctcctccc
     1621 cagtgccttg tctggctacc tctcatgcac aggcttgccc atgttgagaa tgtcttccat
     1681 cccgtggagt gctcctactg ccgatgtgag agtatgatgg gtttccggta ccgatgccag
     1741 cagtgccaca actatcagct ctgccagaat tgcttttggc gtggccatgc cggcggccct
     1801 cacagcaacc agcaccagat gaaggagcat tcctcttgga aatctcctgc aaagaagctg
     1861 agccatgcaa ttagtaaatc tttggggtgt gtacccacga gagaaccccc gcatcctgtt
     1921 tttcctgagc aaccagagaa accacttgac cttgcacata tagttcctcc tcgccctctg
     1981 actaatatga atgacaccat ggttagccac atgtcctctg gagtgcccac tcccaccaag
     2041 aggttacagt atagccagga tatacccagt cacttggccg atgagcatgc gctgatagcc
     2101 tcctatgtgg cccgtctgca gcactgtgca cgtgttctgg acagtcctag ccgactggat
     2161 gaggaacacc gtcttatagc tcgctatgct gcccggctgg ctgcagaagc aggaaacgtg
     2221 actcgtcctc ccactgactt gagctttaac tttgatgcca acaaacaaca aagacagctt
     2281 attgcagaac tggaaaacaa aaacagagag atcctgcagg agattcagcg tctccgcctg
     2341 gaacacgagc aggcctccca gcccacccct gagaaggcac agcagaaccc cacgctgctg
     2401 gcagagctgc ggctgctgag gcaaaggaag gatgaactgg agcagaggat gtcggccctg
     2461 caggagagca ggcgggagct gatggtccag ctggaagagc tgatgaagtt gctgaaggag
     2521 gaagagcaaa agcaggcagc tcaggccaca gggtcaccac atacatcgcc cacccatgga
     2581 ggcggccggc caatgcccat gccagtgcgc tccacgtctg ccggctccac ccccacccac
     2641 tgtccgcagg actcgctgag cggagtcggg ggagacgtgc aggaggcctt cgcacaaggt
     2701 acgaggagaa acctccgcaa tgacctgctg gtggcagctg actccatcac caacaccatg
     2761 tcatccctgg tgaaggagct ccattcagca gaggaaggtg cagaggaaga agaagagaag
     2821 atgcagaatg ggaaagacag aggtctcgag gactacaaag accatgacgg tgattataaa
     2881 gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccgt ttaaacccgc
     2941 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3001 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3061 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3121 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     3181 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     3241 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3301 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3361 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3421 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3481 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3541 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3601 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3661 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3721 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3781 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3841 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3901 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     3961 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     4021 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4081 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4141 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4201 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     4261 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4321 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4381 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4441 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4501 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4561 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4621 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4681 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4741 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4801 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4861 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4921 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4981 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     5041 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5101 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5161 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5221 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5281 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5341 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5401 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5461 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5521 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5581 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5641 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5701 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5761 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5821 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5881 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5941 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6001 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     6061 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6121 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6181 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6241 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6301 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6361 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6421 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6481 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6541 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6601 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6661 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6721 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6781 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6841 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6901 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6961 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7021 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7081 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7141 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7201 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7261 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7321 gcgcacattt ccccgaaaag tgccacctga cgtc
//
