LOCUS       V036681                 7397 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036681
VERSION     V036681
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             983..1687
                     /label="Tnf(NM_013693)"
                     /note="Tnf(NM_013693)"
                     /gene="Tnf"
     CDS             2888..2917
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     polyA_signal    2964..3188
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3234..3662
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3676..4005
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4072..4866
                     /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    5040..5161
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5210..5226)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5234..5250
                     /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(5258..5288)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5303..5324
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5612..6197)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6368..7228)
                     /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(7229..7333)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg tgcctctctc cggttaaagg cgcaaagctc atcctgatct ttctgttcct
      961 aggagccgtt cagtccaatg caatgagcac agaaagcatg atccgcgacg tggaactggc
     1021 agaagaggca ctcccccaaa agatgggggg cttccagaac tccaggcggt gcctatgtct
     1081 cagcctcttc tcattcctgc ttgtggcagg ggccaccacg ctcttctgtc tactgaactt
     1141 cggggtgatc ggtccccaaa gggatgagaa gttcccaaat ggcctccctc tcatcagttc
     1201 tatggcccag accctcacac tcagatcatc ttctcaaaat tcgagtgaca agcctgtagc
     1261 ccacgtcgta gcaaaccacc aagtggagga gcagctggag tggctgagcc agcgcgccaa
     1321 cgccctcctg gccaacggca tggatctcaa agacaaccaa ctagtggtgc cagccgatgg
     1381 gttgtacctt gtctactccc aggttctctt caagggacaa ggctgccccg actacgtgct
     1441 cctcacccac accgtcagcc gatttgctat ctcataccag gagaaagtca acctcctctc
     1501 tgccgtcaag agcccctgcc ccaaggacac ccctgagggg gctgagctca aaccctggta
     1561 tgagcccata tacctgggag gagtcttcca gctggagaag ggggaccaac tcagcgctga
     1621 ggtcaatctg cccaagtact tagactttgc ggagtccggg caggtctact ttggagtcat
     1681 tgctctgggt ggaggcggat caggaggcgg tggatctttg atagttaatt tgacagattc
     1741 aaagggtact tgcctttatg cagaatggga gatgaatttc acaataacat atgaaactac
     1801 aaaccaaacc aataaaacta taaccattgc agtacctgac aaggcgacac acgatggaag
     1861 cagttgtggg gatgaccgga atagtgccaa aataatgata caatttggat tcgctgtctc
     1921 ttgggctgtg aattttacca aggaagcatc tcattattca attcatgaca tcgtgctttc
     1981 ctacaacact agtgatagca cagtatttcc tggtgctgta gctaaaggag ttcatactgt
     2041 taaaaatcct gagaatttca aagttccatt ggatgtcatc tttaagtgca atagtgtttt
     2101 aacttacaac ctgactcctg tcgttcagaa atattggggt attcacctgc aagcttttgt
     2161 ccaaaatggt acagtgagta aaaatgaaca agtgtgtgaa gaagaccaaa ctcccaccac
     2221 tgtggcaccc atcattcaca ccactgcccc gtcgactaca actacactca ctccaacttc
     2281 aacacccact ccaactccaa ctccaactcc aaccgttgga aactacagca ttagaaatgg
     2341 caatactacc tgtctgctgg ctaccatggg gctgcagctg aacatcactg aggagaaggt
     2401 gcctttcatt tttaacatca accctgccac aaccaacttc accggcagct gtcaacctca
     2461 aagtgctcaa cttaggctga acaacagcca aattaagtat cttgacttta tctttgctgt
     2521 gaaaaatgaa aaacggttct atctgaagga agtgaatgtc tacatgtatt tggctaatgg
     2581 ctcagctttc aacatttcca acaagaacct tagcttctgg gatgcccctc tgggaagttc
     2641 ttatatgtgc aacaaagagc aggtgctttc tgtgtctaga gcgtttcaga tcaacacctt
     2701 taacctaaag gtgcaacctt ttaatgtgac aaaaggacag tattctacag ctcaagactg
     2761 cagtgcagat gaagacaact tccttgtgcc catagcggtg ggagcagctc tgggaggagt
     2821 acttattcta gtgttgctgg cttattttat tggtctcaag cgccatcata ctggatatga
     2881 gcaatttgag cagaagctga tctcagagga ggacctgtga ctcgagtcta gagggcccgt
     2941 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3001 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3061 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3121 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     3181 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     3241 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3301 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3361 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3421 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3481 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3541 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3601 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3661 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3721 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3781 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3841 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3901 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     3961 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     4021 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     4081 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4141 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4201 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     4261 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4321 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4381 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4441 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4501 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4561 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     4621 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4681 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4741 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4801 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4861 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4921 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4981 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     5041 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5101 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5161 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     5221 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5281 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5341 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5401 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5461 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5521 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5581 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5641 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5701 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5761 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5821 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5881 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5941 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6001 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6061 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6121 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca
     6181 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6241 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6301 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6361 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6421 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6481 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6541 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6601 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6661 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6721 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6781 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6841 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6901 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6961 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7021 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7081 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7141 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7201 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7261 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7321 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     7381 tcccgatccc ctatggt
//
