LOCUS       V040108                 8446 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040108
VERSION     V040108
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"
     CDS             706..2358
                     /label="RELA(NM_021975)"
                     /note="RELA(NM_021975)"
                     /gene="RELA"
     CDS             2389..3105
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             3112..3138
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3142..3168
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3172..3198
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    3222..3810
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3876..3924
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4126..4554
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4568..4897
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4964..5758
                     /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    5934..6055
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6104..6120)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6128..6144
                     /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(6152..6182)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6197..6218
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6506..7094)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7265..8125)
                     /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(8126..8230)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatgga cgaactgttc
      721 cccctcatct tcccggcaga gccagcccag gcctctggcc cctatgtgga gatcattgag
      781 cagcccaagc agcggggcat gcgcttccgc tacaagtgcg aggggcgctc cgcgggcagc
      841 atcccaggcg agaggagcac agataccacc aagacccacc ccaccatcaa gatcaatggc
      901 tacacaggac cagggacagt gcgcatctcc ctggtcacca aggaccctcc tcaccggcct
      961 cacccccacg agcttgtagg aaaggactgc cgggatggct tctatgaggc tgagctctgc
     1021 ccggaccgct gcatccacag tttccagaac ctgggaatcc agtgtgtgaa gaagcgggac
     1081 ctggagcagg ctatcagtca gcgcatccag accaacaaca accccttcca agttcctata
     1141 gaagagcagc gtggggacta cgacctgaat gctgtgcggc tctgcttcca ggtgacagtg
     1201 cgggacccat caggcaggcc cctccgcctg ccgcctgtcc tttctcatcc catctttgac
     1261 aatcgtgccc ccaacactgc cgagctcaag atctgccgag tgaaccgaaa ctctggcagc
     1321 tgcctcggtg gggatgagat cttcctactg tgtgacaagg tgcagaaaga ggacattgag
     1381 gtgtatttca cgggaccagg ctgggaggcc cgaggctcct tttcgcaagc tgatgtgcac
     1441 cgacaagtgg ccattgtgtt ccggacccct ccctacgcag accccagcct gcaggctcct
     1501 gtgcgtgtct ccatgcagct gcggcggcct tccgaccggg agctcagtga gcccatggaa
     1561 ttccagtacc tgccagatac agacgatcgt caccggattg aggagaaacg taaaaggaca
     1621 tatgagacct tcaagagcat catgaagaag agtcctttca gcggacccac cgacccccgg
     1681 cctccacctc gacgcattgc tgtgccttcc cgcagctcag cttctgtccc caagccagca
     1741 ccccagccct atccctttac gtcatccctg agcaccatca actatgatga gtttcccacc
     1801 atggtgtttc cttctgggca gatcagccag gcctcggcct tggccccggc ccctccccaa
     1861 gtcctgcccc aggctccagc ccctgcccct gctccagcca tggtatcagc tctggcccag
     1921 gccccagccc ctgtcccagt cctagcccca ggccctcctc aggctgtggc cccacctgcc
     1981 cccaagccca cccaggctgg ggaaggaacg ctgtcagagg ccctgctgca gctgcagttt
     2041 gatgatgaag acctgggggc cttgcttggc aacagcacag acccagctgt gttcacagac
     2101 ctggcatccg tcgacaactc cgagtttcag cagctgctga accagggcat acctgtggcc
     2161 ccccacacaa ctgagcccat gctgatggag taccctgagg ctataactcg cctagtgaca
     2221 ggggcccaga ggccccccga cccagctcct gctccactgg gggccccggg gctccccaat
     2281 ggcctccttt caggagatga agacttctcc tccattgcgg acatggactt ctcagccctg
     2341 ctgagtcaga tcagctccgg cggtggagga agcggcggag gtggatctat ggtgagcaag
     2401 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     2461 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     2521 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     2581 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     2641 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     2701 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     2761 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     2821 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     2881 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     2941 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     3001 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     3061 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagctcga gtacccatac
     3121 gacgtacctg attatgcaac ttatccttat gatgtacctg attatgctag ctatccttat
     3181 gatgtcccag actacgcata gtgagggccc gatatctcga caatcaacct ctggattaca
     3241 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     3301 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     3361 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     3421 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     3481 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     3541 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     3601 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga
     3661 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     3721 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     3781 gtcggatctc cctttgggcc gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg
     3841 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     3901 gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     3961 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     4021 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     4081 ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg ccctgtagcg
     4141 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4201 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4261 cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc
     4321 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     4381 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4441 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgggga
     4501 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     4561 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4621 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4681 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4741 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4801 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     4861 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     4921 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     4981 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     5041 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     5101 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     5161 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     5221 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     5281 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     5341 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     5401 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     5461 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     5521 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     5581 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     5641 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     5701 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     5761 gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     5821 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5881 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     5941 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     6001 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     6061 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     6121 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     6181 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     6241 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     6301 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     6361 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     6421 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     6481 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6541 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6601 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6661 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
     6721 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6781 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6841 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6901 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     6961 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     7021 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     7081 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     7141 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     7201 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     7261 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     7321 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     7381 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     7441 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     7501 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     7561 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     7621 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7681 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7741 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7801 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7861 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7921 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7981 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     8041 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     8101 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     8161 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     8221 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     8281 cgatccccta tggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     8341 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     8401 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagg
//
