LOCUS       V038822                 7512 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038822
VERSION     V038822
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..2192
                     /label="FBXL16(NM_153350)"
                     /note="FBXL16(NM_153350)"
                     /gene="FBXL16"
     CDS             2199..2264
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2288..2876
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2942..2990
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3192..3620
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3634..3963
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4030..4824
                     /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    5000..5121
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5170..5186)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5194..5210
                     /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(5218..5248)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5263..5284
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5572..6160)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6331..7191)
                     /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(7192..7296)
                     /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 ccaccatgtc gagcccgggc atcgacggcg
      781 accccaagcc tccatgcttg cctcgaaacg gtctggtgaa gctgccgggc cagcccaacg
      841 gcctgggtgc ggccagcatc accaagggca cgccagccac caagaaccgc ccctgccagc
      901 caccaccccc acccaccctc ccaccaccca gcctggctgc tccactgtcc cgggctgccc
      961 tggctggggg cccgtgcacc ccggcaggtg gaccagcctc agccttggca cctgggcacc
     1021 cagcggagcg gccgccgctg gccacggacg agaagatcct caatgggctc ttctggtatt
     1081 tctcggcctg cgagaagtgt gtgctggccc aggtgtgcaa ggcctggcgg cgcgtgctgt
     1141 accagcccaa gttctgggca ggcctcacgc cggtgctgca tgccaaggag ctctacaacg
     1201 tgctgcctgg tggcgagaag gagttcgtga acctgcaggg ttttgccgcc agaggcttcg
     1261 agggcttctg cctggttggc gtctccgacc tggacatctg tgagttcatt gacaactatg
     1321 cgctctccaa gaagggtgtc aaagccatga gcctcaagcg ctccaccatc acggacgcag
     1381 gcctcgaggt tatgcttgaa cagatgcagg gcgtggtgcg tctggagctg tcgggctgca
     1441 acgacttcac cgaggccggg ctgtggtcca gcctgagcgc gcgcatcacc tcgctgagcg
     1501 tgagtgactg catcaacgtg gccgacgacg ccatcgcggc catctcgcag ctgctgccca
     1561 acctggcgga gctgagcctg caggcctacc acgtgacgga cacggcgctg gcctacttca
     1621 cggcgcgcca gggccacagc acgcacacgc tgcgcctgct ctcctgctgg gagatcacca
     1681 accacggcgt ggtcaacgtg gtgcacagcc tgcccaacct caccgcgctc agcctctcgg
     1741 gctgctccaa ggtcaccgac gacggcgtgg agctcgtggc cgagaacctg cgcaagctgc
     1801 gcagccttga cctctcgtgg tgcccacgca tcaccgacat ggcgctggag tacgtggcct
     1861 gcgacctgca ccgcctagag gagctcgtgc tcgacaggtg tgtacgcatc acggacactg
     1921 gcctcagcta tctgtccacc atgtcgtccc tccgcagcct ctacctgcga tggtgctgcc
     1981 aggtgcaaga cttcgggctg aagcacctcc tggccctggg gagtttgcgc ctcctgtctc
     2041 tggcaggctg cccgctgctc accaccaccg ggctgtcggg cctggtgcag ctgcaggagc
     2101 tggaggagct ggagctgacc aactgccccg gggccacccc cgagctcttc aagtatttct
     2161 cgcagcacct gccccgctgc ctcgtcattg agggatccga ctacaaagac catgacggtg
     2221 attataaaga tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgata
     2281 tctcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2341 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     2401 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     2461 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     2521 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     2581 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     2641 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     2701 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     2761 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     2821 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     2881 aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     2941 acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg
     3001 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     3061 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     3121 cgcagccaac gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag
     3181 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3241 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3301 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
     3361 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3421 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3481 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3541 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
     3601 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3661 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3721 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3781 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3841 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3901 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3961 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     4021 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4081 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4141 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4201 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4261 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4321 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4381 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4441 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4501 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4561 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4621 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4681 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4741 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4801 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc
     4861 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4921 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4981 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     5041 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5101 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     5161 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5221 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5281 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5341 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5401 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5461 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5521 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5581 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5641 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5701 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5761 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5821 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5881 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5941 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6001 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6061 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6121 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6181 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6241 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6301 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6361 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6421 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6481 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6541 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6601 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6661 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6721 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6781 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6841 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6901 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6961 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7021 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7081 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7141 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7201 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7261 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7321 tgacgtcgac ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc
     7381 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     7441 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     7501 aatctgctta gg
//
