LOCUS       V026383                 6540 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026383
VERSION     V026383
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..1220
                     /label="Fgf1(NM_010197)"
                     /note="Fgf1(NM_010197)"
                     /gene="Fgf1"
     CDS             1227..1292
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1316..1904
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    1970..2018
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2220..2648
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2662..2991
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3058..3852
                     /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    4028..4149
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4198..4214)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4222..4238
                     /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(4246..4276)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4291..4312
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4600..5188)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5359..6219)
                     /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(6220..6324)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggc tgaaggggag atcacaacct
      781 tcgcagccct gaccgagagg ttcaacctgc ctctaggaaa ctacaaaaag cccaaactgc
      841 tctactgcag caacgggggc cacttcttga ggatccttcc tgatggcacc gtggatggga
      901 caagggacag gagcgaccag cacattcagc tgcagctcag tgcggaaagt gcgggcgaag
      961 tgtatataaa gggtacggag accggccagt acttggccat ggacaccgaa gggcttttat
     1021 acggctcgca gacaccaaat gaggaatgtc tgttcctgga aaggctggaa gaaaaccatt
     1081 ataacactta cacctccaag aagcatgcgg agaagaactg gtttgtgggc ctcaagaaga
     1141 acgggagctg taagcgcggt cctcggactc actatggcca gaaagccatc ttgtttctgc
     1201 ccctcccggt gtcttctgac ggatccgact acaaagacca tgacggtgat tataaagatc
     1261 atgacatcga ctacaaggat gacgatgaca agtagtgagg gcccgatatc tcgacaatca
     1321 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     1381 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     1441 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     1501 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     1561 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     1621 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     1681 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     1741 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     1801 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     1861 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg
     1921 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     1981 agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     2041 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     2101 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     2161 cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca
     2221 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2281 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2341 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag
     2401 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     2461 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2521 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     2581 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     2641 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     2701 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     2761 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     2821 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     2881 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     2941 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     3001 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     3061 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3121 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     3181 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     3241 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     3301 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     3361 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     3421 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     3481 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     3541 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     3601 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     3661 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     3721 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     3781 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     3841 gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct
     3901 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     3961 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4021 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4081 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4141 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     4201 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4261 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4321 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4381 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4441 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4501 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4561 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4621 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4681 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4741 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     4801 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4861 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4921 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4981 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5041 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5101 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5161 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5221 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5281 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5341 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5401 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5461 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5521 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5581 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5641 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5701 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5761 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5821 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5881 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5941 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6001 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6061 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6121 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6181 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6241 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6301 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     6361 atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca
     6421 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     6481 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
