LOCUS       V020199                 7475 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020199
VERSION     V020199
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             683..2155
                     /label="FGB(NM_005141)"
                     /note="FGB(NM_005141)"
                     /gene="FGB"
     CDS             2162..2227
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2251..2839
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2905..2953
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3155..3583
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3597..3926
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3993..4787
                     /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    4963..5084
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5133..5149)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5157..5173
                     /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(5181..5211)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5226..5247
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5535..6123)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6294..7154)
                     /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(7155..7259)
                     /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 ggatccgcca ccatgaaaag gatggtttct tggagcttcc acaaacttaa
      721 aaccatgaaa catctattat tgctactatt gtgtgttttt ctagttaagt cccaaggtgt
      781 caacgacaat gaggagggtt tcttcagtgc ccgtggtcat cgaccccttg acaagaagag
      841 agaagaggct cccagcctga ggcctgcccc accgcccatc agtggaggtg gctatcgggc
      901 tcgtccagcc aaagcagctg ccactcaaaa gaaagtagaa agaaaagccc ctgatgctgg
      961 aggctgtctt cacgctgacc cagacctggg ggtgttgtgt cctacaggat gtcagttgca
     1021 agaggctttg ctacaacagg aaaggccaat cagaaatagt gttgatgagt taaataacaa
     1081 tgtggaagct gtttcccaga cctcctcttc ttcctttcag tacatgtatt tgctgaaaga
     1141 cctgtggcaa aagaggcaga agcaagtaaa agataatgaa aatgtagtca atgagtactc
     1201 ctcagaactg gaaaagcacc aattatatat agatgagact gtgaatagca atatcccaac
     1261 taaccttcgt gtgcttcgtt caatcctgga aaacctgaga agcaaaatac aaaagttaga
     1321 atctgatgtc tcagctcaaa tggaatattg tcgcacccca tgcactgtca gttgcaatat
     1381 tcctgtggtg tctggcaaag aatgtgagga aattatcagg aaaggaggtg aaacatctga
     1441 aatgtatctc attcaacctg acagttctgt caaaccgtat agagtatact gtgacatgaa
     1501 tacagaaaat ggaggatgga cagtgattca gaaccgtcaa gacggtagtg ttgactttgg
     1561 caggaaatgg gatccatata aacagggatt tggaaatgtt gcaaccaaca cagatgggaa
     1621 gaattactgt ggcctaccag gtgaatattg gcttggaaat gataaaatta gccagcttac
     1681 caggatggga cccacagaac ttttgataga aatggaggac tggaaaggag acaaagtaaa
     1741 ggctcactat ggaggattca ctgtacagaa tgaagccaac aaataccaga tctcagtgaa
     1801 caaatacaga ggaacagccg gtaatgccct catggatgga gcatctcagc tgatgggaga
     1861 aaacaggacc atgaccattc acaacggcat gttcttcagc acgtatgaca gagacaatga
     1921 cggctggtta acatcagatc ccagaaaaca gtgttctaaa gaagacggtg gtggatggtg
     1981 gtataataga tgtcatgcag ccaatccaaa cggcagatac tactggggtg gacagtacac
     2041 ctgggacatg gcaaagcatg gcacagatga tggtgtagta tggatgaatt ggaaggggtc
     2101 atggtactca atgaggaaga tgagtatgaa gatcaggccc ttcttcccac agcaactcga
     2161 ggactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga
     2221 tgacaagtag tgagggcccg atatctcgac aatcaacctc tggattacaa aatttgtgaa
     2281 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     2341 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     2401 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     2461 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     2521 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     2581 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     2641 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     2701 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     2761 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     2821 ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg aaggagacaa
     2881 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt
     2941 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     3001 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     3061 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca
     3121 gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     3181 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3241 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3301 ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3361 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3421 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3481 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat
     3541 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     3601 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3661 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3721 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3781 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3841 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     3901 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     3961 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4021 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4081 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4141 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     4201 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4261 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4321 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4381 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4441 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4501 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     4561 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     4621 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     4681 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     4741 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     4801 gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     4861 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     4921 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     4981 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     5041 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     5101 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     5161 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     5221 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5281 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5341 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5401 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5461 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5521 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5581 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5641 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5701 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     5761 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5821 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5881 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5941 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     6001 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6061 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6121 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6181 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6241 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6301 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6361 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6421 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6481 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6541 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6601 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6661 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6721 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6781 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6841 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6901 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6961 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7021 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7081 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7141 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7201 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7261 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
     7321 ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     7381 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
     7441 gcttgaccga caattgcatg aagaatctgc ttagg
//
