LOCUS       V020411                 9344 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020411
VERSION     V020411
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..4024
                     /label="FNIP2(NM_020840)"
                     /note="FNIP2(NM_020840)"
                     /gene="FNIP2"
     CDS             4031..4096
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4120..4708
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4774..4822
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5024..5452
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5466..5795
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5862..6656
                     /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    6832..6953
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7002..7018)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7026..7042
                     /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(7050..7080)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7095..7116
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7404..7992)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8163..9023)
                     /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(9024..9128)
                     /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 ccatggcccc gaccctgctc cagaagctct tcaacaaaag
      721 gggcagcagc ggcagctccg cggcggcgtc tgcccagggc agggctccta aggaaggacc
      781 cgcctttagt tggtcatgtt cggagtttga cctgaatgag attcgcctga tagtttacca
      841 ggactgtgac aggagaggca gacaagtctt gtttgactct aaagctgttc aaaagattga
      901 ggaggtgaca gctcagaaaa cagaggatgt tcctattaaa atatcagcca agtgctgcca
      961 gggaagcagc agtgtcagca gcagtagcag cagcagcatc tcttcccaca gttcttctgg
     1021 gggatcttca catcatgcta aggaacagct tccaaagtac cagtacacaa gaccagcttc
     1081 cgatgtcaac atgttagggg aaatgatgtt tggctcagtt gccatgagtt acaaaggctc
     1141 caccttaaag atacactaca tacgttctcc tccacaactg atgattagta aagtcttctc
     1201 tgctagaatg ggcagcttct gtggaagtac aaataacttg caagacagct ttgagtacat
     1261 caaccaagat cctaatttgg gaaaactgaa cacaaatcaa aatagtttgg gtccttgtcg
     1321 tactggaagt aacctagcac acagcacacc agttgatatg ccaagcagag gacagaatga
     1381 agacagggac agtggcattg ctcgatcagc ctcactaagc agtcttttga tcacaccttt
     1441 cccatctcca agctcctcta catcttcttc cagcagttac cagcgccgct ggcttcgaag
     1501 tcagacaaca agtttggaaa atggcatcat cccaagaagg tcaactgatg agacattcag
     1561 cttggctgaa gaaacctgta gctctaatcc agctatggtt aggaggaaga aaattgccat
     1621 aagcatcatc ttttccctat gtgagaaaga agaagcacaa aggaatttcc aggacttctt
     1681 cttttctcat tttcccctgt ttgaatctca catgaacagg ctgaagagtg caattgaaaa
     1741 ggctatgatc tcctgtagga aaatagcaga atcaagtctc cgagtccagt tttatgtcag
     1801 ccgtttgatg gaagctctgg gagaattcag aggaactatc tggaacttat attctgttcc
     1861 aaggatagct gaacctgtat ggcttactat gatgtccggc actttggaaa aaaaccagct
     1921 ctgccagcgc tttctcaagg agtttacact tctgatagaa cagataaata aaaaccagtt
     1981 ttttgctgcc ttactgactg cggtgttaac ctaccacctg gcctgggtcc caactgtcat
     2041 gcctgtggat caccctccca tcaaagcctt ctcagagaaa cgtacctccc agtcagtgaa
     2101 catgctggcc aaaacacatc cgtataatcc tctttgggca cagctgggtg atctttacgg
     2161 agccataggc tctccagtga gactgactcg caccgtagtg gtagggaagc agaaggactt
     2221 agtccagcga atactttatg tcctgaccta ctttctccgt tgctctgagc tacaagagaa
     2281 ccagctgacc tggagtggca atcatggtga aggtgaccaa gttttaaatg ggagcaagat
     2341 cataacagcc ttggagaaag gagaggtgga ggagtctgag tatgtggtca ttacggtgag
     2401 gaacgagccc gctcttgtac cccccatcct accaccaaca gcagcagaga gacacaaccc
     2461 ctggccgaca gggtttcctg agtgcccaga gggcactgac agtagagacc tgggtcttaa
     2521 acctgacaaa gaagctaaca ggaggccaga gcagggttct gaggcttgca gcgcagggtg
     2581 cctggggcca gcatcagacg cttcctggaa acctcagaat gcattttgtg gggatgagaa
     2641 aaataaagag gcaccgcaag atggctcttc aagacttccc agctgtgaag ttttgggggc
     2701 aggaatgaag atggaccagc aagctgtctg tgagctgttg aaagtggaga tgcctacaag
     2761 actgccagac cggtcagtgg cctggccttg ccctgacaga catctccggg agaaaccttc
     2821 cttagaaaag gtcactttcc agattggaag ctttgcatct ccagagtctg actttgaaag
     2881 ccgcatgaaa aaaatggagg aacgggtgaa ggcctgtggc ccctccttgg aggccagtga
     2941 ggctgctgat gtggctcagg acccgcaggt ttctaggagc ccttttaaac ctggctttca
     3001 ggagaatgtt tgctgtcctc agaatcggct ttcagagggg gatgaaggcg agtctgacaa
     3061 gggttttgca gaggacagag gcagcagaaa cgacatggca gcagatattg ctgggcagct
     3121 cagccacgct gctgacttgg gcacagcctc ccacggtgca ggaggaacgg gagggaggag
     3181 gctggaggcc actagaggtt tgtatgtgaa ggctgcggaa ggacctgtgc tggagcctgt
     3241 tgcccccagg tgtgtccagc ggggccctgg cctcgtggct ggtgcgaata tcccctgtgg
     3301 ggatgacaac aagaaggcca acttcaggac tgaaggagac attccccgaa atgaaagctc
     3361 agatagcgcc ctgggagaca gtgacgacga agcctgcgct tcagccatgc tagatctggg
     3421 tcacggtggt gacaggactg gagggtcctt ggaagtggag ctgcctctgc caaggtctca
     3481 gagcatcagc acccagaatg taaggaactt tggccgctca cttctggcgg gctactgccc
     3541 cacatacatg cctgatcttg tgcttcatgg gaccggcagt gatgagaagc tgaagcagtg
     3601 cctggtggcc gaccttgtcc acacagtcca tcatccagtc ctggatgagc caatagctga
     3661 agctgtctgt attatcgcag acacggataa atggagtgtg caggtagcta caagtcagag
     3721 gaaagtgacg gacaacatga aactaggcca ggatgtcctg gtctctagtc aggtgtccag
     3781 tttgcttcag tccattttac agctctataa gcttcacctc cctgctgatt tttgcatcat
     3841 gcatcttgaa gatagactac aggagatgta ccttaaaagt aaaatgctat ctgaatatct
     3901 ccggggacac acacgagtcc atgtgaaaga attaggtgtc gtactgggga ttgaatccaa
     3961 cgacctgcct ctgttgactg ctattgccag tactcattct ccttatgtgg ctcaaatact
     4021 cttactcgag gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa
     4081 ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct ggattacaaa
     4141 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     4201 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     4261 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     4321 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     4381 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     4441 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     4501 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     4561 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     4621 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     4681 cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga
     4741 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     4801 acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     4861 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     4921 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     4981 gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     5041 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     5101 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     5161 cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc
     5221 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     5281 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     5341 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt
     5401 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     5461 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5521 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     5581 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     5641 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     5701 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     5761 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     5821 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     5881 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     5941 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     6001 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     6061 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     6121 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     6181 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     6241 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     6301 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     6361 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     6421 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     6481 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     6541 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     6601 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     6661 gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     6721 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     6781 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     6841 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     6901 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     6961 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     7021 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     7081 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     7141 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     7201 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     7261 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     7321 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     7381 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     7441 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     7501 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     7561 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     7621 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     7681 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     7741 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     7801 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     7861 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     7921 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     7981 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     8041 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     8101 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     8161 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     8221 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     8281 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     8341 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     8401 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     8461 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     8521 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     8581 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     8641 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     8701 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     8761 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     8821 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     8881 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     8941 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     9001 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     9061 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     9121 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg
     9181 atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     9241 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
     9301 caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
