LOCUS       V018660                 7259 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018660
VERSION     V018660
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             671..1939
                     /label="TNFRSF19(NM_018647)"
                     /note="TNFRSF19(NM_018647)"
                     /gene="TNFRSF19"
     CDS             1946..2011
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2035..2623
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2689..2737
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2939..3367
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3381..3710
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3777..4571
                     /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    4747..4868
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4917..4933)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4941..4957
                     /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(4965..4995)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5010..5031
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5319..5907)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6078..6938)
                     /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(6939..7043)
                     /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 taccgccacc atggctttaa aagtgctact agaacaagag aaaacgtttt tcactctttt
      721 agtattacta ggctatttgt catgtaaagt gacttgtgaa tcaggagact gtagacagca
      781 agaattcagg gatcggtctg gaaactgtgt tccctgcaac cagtgtgggc caggcatgga
      841 gttgtctaag gaatgtggct tcggctatgg ggaggatgca cagtgtgtga cgtgccggct
      901 gcacaggttc aaggaggact ggggcttcca gaaatgcaag ccctgtctgg actgcgcagt
      961 ggtgaaccgc tttcagaagg caaattgttc agccaccagt gatgccatct gcggggactg
     1021 cttgccagga ttttatagga agacgaaact tgtcggcttt caagacatgg agtgtgtgcc
     1081 ttgtggagac cctcctcctc cttacgaacc gcactgtgcc agcaaggtca acctcgtgaa
     1141 gatcgcgtcc acggcctcca gcccacggga cacggcgctg gctgccgtta tctgcagcgc
     1201 tctggccacc gtcctgctgg ccctgctcat cctctgtgtc atctattgta agagacagtt
     1261 tatggagaag aaacccagct ggtctctgcg gtcacaggac attcagtaca acggctctga
     1321 gctgtcgtgt tttgacagac ctcagctcca cgaatatgcc cacagagcct gctgccagtg
     1381 ccgccgtgac tcagtgcaga cctgcgggcc ggtgcgcttg ctcccatcca tgtgctgtga
     1441 ggaggcctgc agccccaacc cggcgactct tggttgtggg gtgcattctg cagccagtct
     1501 tcaggcaaga aacgcaggcc cagccgggga gatggtgccg actttcttcg gatccctcac
     1561 gcagtccatc tgtggcgagt tttcagatgc ctggcctctg atgcagaatc ccatgggtgg
     1621 tgacaacatc tctttttgtg actcttatcc tgaactcact ggagaagaca ttcattctct
     1681 caatccagaa cttgaaagct caacgtcttt ggattcaaat agcagtcaag atttggttgg
     1741 tggggctgtt ccagtccagt ctcattctga aaactttaca gcagctactg atttatctag
     1801 atataacaac acactggtag aatcagcatc aactcaggat gcactaacta tgagaagcca
     1861 gctagatcag gagagtggtg ctgtcatcca cccagccact cagacgtccc tccaggtaag
     1921 gcagcgactg ggttccctgc tcgaggacta caaagaccat gacggtgatt ataaagatca
     1981 tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct cgacaatcaa
     2041 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     2101 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     2161 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     2221 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     2281 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     2341 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     2401 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt
     2461 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     2521 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     2581 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa cgggggaggc
     2641 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     2701 gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     2761 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     2821 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     2881 ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac
     2941 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3001 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3061 ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt
     3121 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     3181 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3241 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     3301 gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3361 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3421 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3481 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3541 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3601 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3661 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     3721 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     3781 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3841 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3901 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     3961 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4021 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4081 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4141 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4201 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4261 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     4321 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4381 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4441 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4501 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4561 agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg
     4621 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4681 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4741 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4801 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4861 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     4921 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4981 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5041 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5101 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5161 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5221 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5281 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5341 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5401 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5461 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     5521 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5581 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5641 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5701 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     5761 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5821 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5881 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5941 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6001 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6061 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6121 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6181 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6241 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6301 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6361 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6421 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6481 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6541 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6601 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6661 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6721 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6781 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6841 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6901 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6961 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7021 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     7081 tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat
     7141 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
     7201 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttagg
//
