LOCUS       V046703                 8621 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046703
VERSION     V046703
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             890..934
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic
                     ribonuclease A"
                     /transl_table=1
                     /translation="KETAAAKFERQHMDS"
                     /label="S-Tag"
     CDS             941..1054
                     /codon_start=1
                     /note="selected from a peptide library; binds streptavidin
                     with nanomolar affinity (Keefe et al., 2001)"
                     /product="streptavidin-binding peptide"
                     /transl_table=1
                     /translation="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
                     /label="SBP"
     CDS             1061..1087
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             1094..3394
                     /label="NSUN2(NM_017755)"
                     /note="NSUN2(NM_017755)"
                     /gene="NSUN2"
     CDS             3425..4144
                     /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"
     polyA_signal    4188..4412
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4458..4886
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4900..5229
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5296..6090
                     /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    6264..6385
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6434..6450)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6458..6474
                     /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(6482..6512)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6527..6548
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6836..7421)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7592..8452)
                     /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(8453..8557)
                     /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 cttaccatga aagaaaccgc
      901 tgctgctaaa ttcgaacgcc agcacatgga cagcggtgga atggacgaga agaccaccgg
      961 ctggagaggc ggccacgtgg tggagggcct ggccggcgag ctggagcagc tgagagccag
     1021 actggagcac caccctcagg gccagagaga acctggtacc tacccatacg atgttccaga
     1081 ttacgctgga tccatggggc ggcggtcgcg gggtcggcgg ctccagcaac agcagcggcc
     1141 ggaggacgcg gaggatggcg ccgagggtgg tggaaagcgc ggcgaggcgg gctgggaagg
     1201 aggctacccc gagatcgtca aggagaacaa gctgttcgag cactactacc aggagctcaa
     1261 gatcgtgccc gagggcgagt ggggccagtt catggacgct ctcagggagc cgctcccggc
     1321 cactttaaga attactggtt acaaaagcca cgcaaaagag attctccatt gcttaaagaa
     1381 caaatatttt aaggaattgg aggacctgga ggtggacggt cagaaagttg aagttccaca
     1441 gccactgagt tggtatcctg aagaacttgc ctggcacaca aatttaagtc gaaaaatctt
     1501 gagaaaatcg ccacacttgg aaaagtttca tcagtttcta gttagtgaaa cagaatctgg
     1561 aaatattagt cgtcaagaag ctgttagcat gatcccacca ctgctcctca acgtgcggcc
     1621 tcatcataag atcttagata tgtgtgcagc acctggctca aagaccacac agttaattga
     1681 aatgctacat gccgacatga atgtcccctt tccagaggga tttgttattg cgaatgatgt
     1741 ggacaacaag cgctgctacc tgctcgtcca tcaagccaag aggctgagca gcccctgcat
     1801 catggtggtc aaccatgatg cctccagcat acccaggctc cagatagatg tggacggcag
     1861 gaaagagatc ctcttctatg atcgaatttt atgtgatgtc ccttgcagtg gagacggcac
     1921 tatgagaaaa aacattgatg tttggaaaaa gtggaccacc ttaaatagct tgcagctaca
     1981 tggcttacag ctgcggattg caacacgcgg ggctgaacag ctggctgaag gtggaaggat
     2041 ggtgtattcc acgtgttcac taaaccctat tgaggatgaa gcagtcatag catctttact
     2101 ggaaaaaagt gaaggtgctt tggagcttgc tgatgtgtct aatgaactgc cagggctgaa
     2161 gtggatgcct ggaatcacac agtggaaggt aatgacgaaa gatgggcagt ggtttacaga
     2221 ctgggacgct gttcctcaca gcagacacac ccagatccga cctaccatgt tccctccgaa
     2281 ggacccagaa aagctgcagg ccatgcacct ggagcgatgc cttaggatat taccccatca
     2341 tcagaatact ggagggtttt ttgtggcagt attggtgaaa aaatcttcaa tgccgtggaa
     2401 taaacgtcag ccaaagcttc agggtaaatc tgcagagacc agagaaagca cacagctgag
     2461 ccctgcagat ctcacagaag ggaaacccac agatccctct aagctggaaa gtccgtcatt
     2521 cacaggaact ggtgacacag aaatagctca tgcaactgag gatttagaga ataatggcag
     2581 taagaaagat ggcgtgtgtg gtcctcctcc atcaaagaaa atgaagttat ttggatttaa
     2641 agaagatcca tttgtattta ttcctgaaga tgacccatta tttccaccta ttgagaaatt
     2701 ttatgctttg gatccttcat tcccaaggat gaatttgtta actcggacta cagaagggaa
     2761 gaaaaggcag ctctacatgg tttctaagga gttgcggaat gtgctgctga ataacagtga
     2821 gaagatgaag gttattaaca cggggatcaa agtctggtgt agaaataaca gcggtgaaga
     2881 gtttgactgt gctttccggc tggcacagga gggaatatat acattgtatc catttattaa
     2941 ctcaagaatt attactgtat caatggaaga tgttaagata ctgttgaccc aggaaaatcc
     3001 cttttttaga aaactcagca gtgagaccta cagtcaagca aaggacctgg caaagggaag
     3061 catcgtgctg aagtatgaac cagattctgc gaatccagac gctctgcagt gtcccatcgt
     3121 cttatgcgga tggcggggaa aggcctccat tcgaactttt gtgcccaaga atgaacggct
     3181 tcattatctc aggatgatgg ggctggaggt attgggagaa aagaagaagg aaggggttat
     3241 cctcacaaat gagagtgcag ccagcaccgg acagccagac aatgacgtga ctgagggaca
     3301 gagagcagga gagcccaaca gcccagatgc agaagaggcc aacagtccag acgtgacagc
     3361 aggctgtgac ccggcggggg tccatccacc ccggtctaga gggcccttcg ggggtggagg
     3421 ctctatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     3481 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     3541 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     3601 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     3661 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     3721 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     3781 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     3841 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     3901 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     3961 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     4021 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     4081 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     4141 gtaactcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     4201 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     4261 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     4321 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     4381 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     4441 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4501 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4561 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4621 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4681 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4741 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4801 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4861 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     4921 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4981 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     5041 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     5101 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     5161 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     5221 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     5281 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     5341 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     5401 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     5461 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     5521 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     5581 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     5641 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     5701 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     5761 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     5821 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     5881 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     5941 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     6001 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     6061 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     6121 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     6181 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     6241 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     6301 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6361 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     6421 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     6481 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     6541 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     6601 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     6661 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     6721 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     6781 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6841 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6901 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6961 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     7021 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     7081 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     7141 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     7201 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     7261 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     7321 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg
     7381 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     7441 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     7501 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     7561 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     7621 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     7681 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     7741 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     7801 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7861 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7921 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7981 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     8041 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     8101 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     8161 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     8221 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     8281 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     8341 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     8401 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     8461 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     8521 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     8581 ctgacgtcga cggatcggga gatctcccga tcccctatgg t
//
