LOCUS       V019259                 8372 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019259
VERSION     V019259
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..3052
                     /label="UFL1(NM_015323)"
                     /note="UFL1(NM_015323)"
                     /gene="UFL1"
     CDS             3059..3124
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3148..3736
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3802..3850
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4052..4480
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4494..4823
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4890..5684
                     /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    5860..5981
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6030..6046)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6054..6070
                     /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(6078..6108)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6123..6144
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6432..7020)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7191..8051)
                     /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(8052..8156)
                     /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 atggcggacg cctgggaaga gattaggcgg ttggcggccg acttccagcg
      721 ggcgcagttc gccgaggcca cgcagaggtt gtccgagcgg aactgcattg agattgttaa
      781 taaattgatt gctcagaaac agctagaagt agttcataca ctcgatggaa aggaatatat
      841 tactccagcc caaattagta aagaaatgag agatgagcta catgtccgag gtggtcgagt
      901 aaacattgtt gatctacaac aggtaattaa tgtggacctg attcatattg aaaatagaat
      961 tggtgacatt attaaatcag aaaagcatgt tcagttagtg ttgggacaac tgatagatga
     1021 gaattatttg gatcggttgg cagaagaggt caatgataaa ttgcaagaaa gtggtcaggt
     1081 caccatatca gaactgtgta aaacttatga tcttcctggg aactttctga cacaggcact
     1141 aactcagcga cttggtagaa ttatcagtgg acatattgat cttgataata gaggagtaat
     1201 ttttacggaa gcttttgtag ctcgacataa agcacgtatc cgtggactat tcagtgctat
     1261 tacccggcct acagctgtga attctttgat ttcaaaatat ggatttcagg agcagcttct
     1321 ttactctgtg cttgaggaac ttgttaatag cggacgctta cgaggcactg tggttggtgg
     1381 gagacaggat aaagctgtgt ttgtccctga catctactcc aggacacaga gtacttgggt
     1441 ggattccttt ttcaggcaga atggctatct agaatttgat gctttgtcca gacttggaat
     1501 cccagatgct gtaagctaca taaagaaaag atataagact acacaactct tgtttttgaa
     1561 agcagcttgt gttggtcaag gacttgtgga tcaagtggaa gcatcagtag aagaagccat
     1621 cagctctgga acatgggttg atattgcacc tctgctaccc acttctttat cagttgaaga
     1681 tgctgccata ttgcttcagc aggtgatgag ggcattcagc aaacaggcct caactgtagt
     1741 ctttagcgac actgttgtag tcagtgaaaa atttataaat gactgtacag aactgttccg
     1801 tgagctgatg caccagaaag ctgaaaagga aatgaaaaat aatcctgtgc atttaatcac
     1861 tgaagaagat ctgaaacaaa tctccacttt agaaagcgtt agtacaagta aaaaggataa
     1921 aaaagatgag cgaagaagga aagcaacaga gggcagtgga agcatgagag gaggaggtgg
     1981 gggcaatgcc agagagtaca aaattaaaaa agtcaagaag aaaggaagaa aagatgatga
     2041 tagtgatgat gaatctcaat catcccacac tggaaagaag aagccagaga tcagttttat
     2101 gttccaggat gagattgaag attttttaag aaaacacata caagatgccc ctgaggagtt
     2161 tatttcggaa cttgctgagt acttaataaa acctcttaat aaaacttatc tcgaggtggt
     2221 acgttcagta ttcatgtctt caacaacttc tgcttctggg acgggcagaa aacgcacaat
     2281 caaggacttg caagaagaag tttcaaacct gtacaataac attaggttat ttgaaaaagg
     2341 gatgaagttt tttgcagatg acacacaggc tgctcttacc aaacacttgc tgaagtcagt
     2401 gtgtactgat atcactaacc tcattttcaa cttcttagct tcggatttaa tgatggcagt
     2461 agacgatcct gcagccatta caagtgaaat aagaaagaaa attttaagta aattatcaga
     2521 agaaaccaaa gtagctctta caaaactcca taactctctg aatgaaaaga gcatagaaga
     2581 ctttatttct tgtctggatt ctgcagcaga agcttgtgat attatggtga aaaggggaga
     2641 caaaaaaagg gaaagacaga tactgttcca acatcgacaa gcactggctg aacagctaaa
     2701 ggtcacagaa gaccctgctc ttattctgca cctcacatca gtcctgttgt ttcagttttc
     2761 aacccacagc atgctccatg cacctggaag atgtgtccca cagatcattg cttttcttaa
     2821 tagtaaaatt ccagaggatc agcatgctct tttggtaaag tatcaaggtt tggttgtaaa
     2881 gcagctagtc agtcaaagta agaagactgg gcagggagat tatcccttga ataatgaatt
     2941 agacaaagaa caagaagatg ttgccagtac tactcgtaaa gagcttcaag aactttcttc
     3001 atccattaaa gaccttgttc tcaaatctag gaaatcatct gtgacggaag agctcgagga
     3061 ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga
     3121 caagtagtga gggcccgata tctcgacaat caacctctgg attacaaaat ttgtgaaaga
     3181 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     3241 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     3301 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     3361 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     3421 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     3481 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     3541 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     3601 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     3661 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     3721 tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag gagacaatac
     3781 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac gggtgttggg
     3841 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     3901 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     3961 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcagat
     4021 ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     4081 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4141 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4201 aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4261 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4321 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4381 aaccctatct cggtctattc ttttgattta taagggattt tggggatttc ggcctattgg
     4441 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     4501 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4561 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4621 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4681 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4741 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4801 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     4861 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4921 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4981 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     5041 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     5101 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     5161 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     5221 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     5281 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5341 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     5401 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5461 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5521 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5581 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     5641 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     5701 cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     5761 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     5821 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     5881 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     5941 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     6001 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     6061 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     6121 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     6181 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     6241 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     6301 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6361 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6421 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6481 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6541 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6601 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     6661 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6721 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6781 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6841 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     6901 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6961 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7021 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7081 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     7141 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     7201 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     7261 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7321 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7381 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7441 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7501 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7561 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     7621 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7681 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7741 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7801 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7861 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7921 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7981 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     8041 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     8101 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     8161 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     8221 cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     8281 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     8341 tgaccgacaa ttgcatgaag aatctgctta gg
//
