LOCUS       V021799                 6746 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021799
VERSION     V021799
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             941..2236
                     /label="Fgl2(NM_008013)"
                     /note="Fgl2(NM_008013)"
                     /gene="Fgl2"
     CDS             2243..2308
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2346..2570
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2616..3044
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3058..3387
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3454..4248
                     /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    4422..4543
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4592..4608)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4616..4632
                     /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(4640..4670)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4685..4706
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4994..5579)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5750..6610)
                     /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(6611..6715)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgaggcttc ctggttggtt
      961 gtggctgagt tctgccgtcc tcgctgcctg ccgagcggtg gaggagcaca acctgactga
     1021 ggggctggag gatgccagcg cccaggctgc ctgccccgcg aggctggagg gcagcgggag
     1081 gtgcgagggg agccagtgcc ccttccagct caccctgccc acgctgacca tccagctccc
     1141 gcggcagctt ggcagcatgg aggaggtgct caaagaagtg cggaccctca aggaagcagt
     1201 ggacagtctg aagaaatcct gccaggactg taagttgcag gctgacgacc atcgagatcc
     1261 cggcgggaat ggagggaatg gagcagagac agccgaggac agtagagtcc aggaactgga
     1321 gagtcaggtg aacaagctgt cctcagagct gaagaatgca aaggaccaga tccaggggct
     1381 gcaggggcgc ctggagacgc tccatctggt aaatatgaac aacattgaga actacgtgga
     1441 caacaaagtg gcaaatctaa ccgttgtggt caacagtttg gatggcaagt gttccaagtg
     1501 tcccagccaa gaacacatgc agtcacagcc ggttcaacat ctaatataca aagattgttc
     1561 cgaccactac gtgctaggaa ggagaagcag tggggcctac agagttaccc ctgatcacag
     1621 aaacagcagc tttgaggtct actgtgacat ggagaccatg ggtggaggct ggacggtgct
     1681 gcaggctcgc cttgatggca gcaccaactt caccagagag tggaaagact acaaagccgg
     1741 ctttggaaac cttgaacgag aattttggtt gggcaacgat aaaattcatc ttctgaccaa
     1801 gagtaaggaa atgattttga gaatagatct tgaagacttt aatggtctca cactttatgc
     1861 cttgtatgat cagttttatg tggctaatga atttctcaaa taccgattac acatcggtaa
     1921 ctacaatggc acggcagggg atgccttgcg tttcagtcga cactacaacc atgacctgag
     1981 gtttttcaca accccagaca gagacaacga tcggtacccc tctgggaact gtgggctcta
     2041 ttacagctca ggctggtggt ttgattcatg tctctctgcc aacttaaatg gcaaatatta
     2101 ccaccagaaa tacaaaggtg tccgtaatgg gattttctgg ggcacctggc ctggtataaa
     2161 ccaggcacag ccaggtggct acaagtcctc cttcaaacag gccaagatga tgattaggcc
     2221 caagaatttc aagccactcg aggactacaa agaccatgac ggtgattata aagatcatga
     2281 catcgactac aaggatgacg atgacaagta gtgagggccc gtttaaaccc gctgatcagc
     2341 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     2401 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     2461 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     2521 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     2581 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     2641 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     2701 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2761 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     2821 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     2881 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     2941 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     3001 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     3061 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3121 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3181 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3241 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3301 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     3361 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     3421 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     3481 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     3541 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     3601 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     3661 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     3721 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     3781 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     3841 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     3901 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     3961 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     4021 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4081 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4141 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4201 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     4261 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     4321 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     4381 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     4441 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4501 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     4561 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4621 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4681 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4741 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4801 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4861 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4921 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4981 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5041 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5101 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5161 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5221 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5281 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5341 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5401 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5461 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5521 ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5581 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5641 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5701 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5761 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5821 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5881 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5941 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6001 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6061 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6121 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6181 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6241 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6301 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6361 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6421 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6481 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6541 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6601 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6661 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6721 ttccccgaaa agtgccacct gacgtc
//
