LOCUS       V028873                 8386 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028873
VERSION     V028873
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..1401
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric far-red fluorescent protein (Shcherbo
                     et al., 2009)"
                     /transl_table=1
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
                     /label="mKate2"
     CDS             1438..3045
                     /label="SRC(NM_005417)"
                     /note="SRC(NM_005417)"
                     /gene="SRC"
     CDS             3052..3078
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3082..3108
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3112..3138
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    3162..3750
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3816..3864
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4066..4494
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4508..4837
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4904..5698
                     /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    5874..5995
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6044..6060)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6068..6084
                     /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(6092..6122)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6137..6158
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6446..7034)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7205..8065)
                     /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(8066..8170)
                     /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 ccaccatggt gagcgagctg
      721 attaaggaga acatgcacat gaagctgtac atggagggca ccgtgaacaa ccaccacttc
      781 aagtgcacat ccgagggcga aggcaagccc tacgagggca cccagaccat gagaatcaag
      841 gcggtcgagg gcggccctct ccccttcgcc ttcgacatcc tggctaccag cttcatgtac
      901 ggcagcaaaa ccttcatcaa ccacacccag ggcatccccg acttctttaa gcagtccttc
      961 cccgagggct tcacatggga gagagtcacc acatacgaag acgggggcgt gctgaccgct
     1021 acccaggaca ccagcctcca ggacggctgc ctcatctaca acgtcaagat cagaggggtg
     1081 aacttcccat ccaacggccc tgtgatgcag aagaaaacac tcggctggga ggcctccacc
     1141 gagaccctgt accccgctga cggcggcctg gaaggcagag ccgacatggc cctgaagctc
     1201 gtgggcgggg gccacctgat ctgcaacttg aagaccacat acagatccaa gaaacccgct
     1261 aagaacctca agatgcccgg cgtctactat gtggacagaa gactggaaag aatcaaggag
     1321 gccgacaaag agacctacgt cgagcagcac gaggtggctg tggccagata ctgcgacctc
     1381 cctagcaaac tggggcacag aggtggaggc ggatcaggag gcggtggatc ttctagaatg
     1441 ggtagcaaca agagcaagcc caaggatgcc agccagcggc gccgcagcct ggagcccgcc
     1501 gagaacgtgc acggcgctgg cgggggcgct ttccccgcct cgcagacccc cagcaagcca
     1561 gcctcggccg acggccaccg cggccccagc gcggccttcg cccccgcggc cgccgagccc
     1621 aagctgttcg gaggcttcaa ctcctcggac accgtcacct ccccgcagag ggcgggcccg
     1681 ctggccggtg gagtgaccac ctttgtggcc ctctatgact atgagtctag gacggagaca
     1741 gacctgtcct tcaagaaagg cgagcggctc cagattgtca acaacacaga gggagactgg
     1801 tggctggccc actcgctcag cacaggacag acaggctaca tccccagcaa ctacgtggcg
     1861 ccctccgact ccatccaggc tgaggagtgg tattttggca agatcaccag acgggagtca
     1921 gagcggttac tgctcaatgc agagaacccg agagggacct tcctcgtgcg agaaagtgag
     1981 accacgaaag gtgcctactg cctctcagtg tctgacttcg acaacgccaa gggcctcaac
     2041 gtgaagcact acaagatccg caagctggac agcggcggct tctacatcac ctcccgcacc
     2101 cagttcaaca gcctgcagca gctggtggcc tactactcca aacacgccga tggcctgtgc
     2161 caccgcctca ccaccgtgtg ccccacgtcc aagccgcaga ctcagggcct ggccaaggat
     2221 gcctgggaga tccctcggga gtcgctgcgg ctggaggtca agctgggcca gggctgcttt
     2281 ggcgaggtgt ggatggggac ctggaacggt accaccaggg tggccatcaa aaccctgaag
     2341 cctggcacga tgtctccaga ggccttcctg caggaggccc aggtcatgaa gaagctgagg
     2401 catgagaagc tggtgcagtt gtatgctgtg gtttcagagg agcccattta catcgtcacg
     2461 gagtacatga gcaaggggag tttgctggac tttctcaagg gggagacagg caagtacctg
     2521 cggctgcctc agctggtgga catggctgct cagatcgcct caggcatggc gtacgtggag
     2581 cggatgaact acgtccaccg ggaccttcgt gcagccaaca tcctggtggg agagaacctg
     2641 gtgtgcaaag tggcggactt tgggctggct cggctcattg aagacaatga gtacacggcg
     2701 cggcaaggtg ccaaattccc catcaagtgg acggctccag aagctgccct ctatggccgc
     2761 ttcaccatca agtcggacgt gtggtccttc gggatcctgc tgactgagct caccacaaag
     2821 ggacgggtgc cctaccctgg gatggtgaac cgcgaggtgc tggaccaggt ggagcggggc
     2881 taccggatgc cctgcccgcc ggagtgtccc gagtccctgc acgacctcat gtgccagtgc
     2941 tggcggaagg agcctgagga gcggcccacc ttcgagtacc tgcaggcctt cctggaggac
     3001 tacttcacgt ccaccgagcc ccagtaccag cccggggaga acctcctcga gtacccatac
     3061 gacgtacctg attatgcaac ttatccttat gatgtacctg attatgctag ctatccttat
     3121 gatgtcccag actacgcata gtgagggccc gatatctcga caatcaacct ctggattaca
     3181 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     3241 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     3301 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     3361 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     3421 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     3481 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     3541 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga
     3601 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     3661 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     3721 gtcggatctc cctttgggcc gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg
     3781 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     3841 gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     3901 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     3961 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     4021 ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg ccctgtagcg
     4081 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4141 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4201 cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc
     4261 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     4321 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4381 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgggga
     4441 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     4501 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4561 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4621 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4681 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4741 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     4801 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     4861 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     4921 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4981 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     5041 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     5101 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     5161 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     5221 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     5281 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     5341 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     5401 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     5461 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     5521 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     5581 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     5641 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     5701 gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     5761 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5821 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     5881 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5941 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     6001 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     6061 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     6121 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     6181 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     6241 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     6301 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     6361 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     6421 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6481 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6541 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6601 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
     6661 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6721 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6781 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6841 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     6901 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6961 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     7021 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     7081 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     7141 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     7201 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     7261 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     7321 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     7381 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     7441 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     7501 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     7561 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7621 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7681 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7741 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7801 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7861 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7921 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7981 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     8041 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     8101 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     8161 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     8221 cgatccccta tggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     8281 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     8341 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagg
//
