LOCUS       V023884                 8162 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023884
VERSION     V023884
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             779..2842
                     /label="USP49(NM_001384542)"
                     /note="USP49(NM_001384542)"
                     /gene="USP49"
     CDS             2849..2914
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2938..3526
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3592..3640
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3842..4270
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4284..4613
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4680..5474
                     /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    5650..5771
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5820..5836)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5844..5860
                     /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(5868..5898)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5913..5934
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6222..6810)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6981..7841)
                     /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(7842..7946)
                     /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 ctcggatcca ctagtccagt gtggtggaat tcgccaccat
      781 ggatagatgc aaacatgtag ggcggttacg gctcgcccag gaccactcca tcctgaaccc
      841 tcagaagtgg tgctgcttag agtgtgccac caccgagtcc gtgtgggcct gcctcaagtg
      901 ctcccacgtg gcctgcggcc gctatattga ggaccacgcc ctgaaacact ttgaggagac
      961 gggacacccg ctagccatgg aagtccggga tctctacgtg ttctgttacc tgtgcaagga
     1021 ctacgtgctc aatgataacc cagaggggga cctgaagctg ctaagaagct ccctcctggc
     1081 ggtccggggc cagaaacagg acacgccggt gagacgtggg cggacgctgc ggtccatggc
     1141 ttcgggtgag gacgtggtcc tgccgcagcg cgctcctcag ggacagccgc agatgctcac
     1201 ggctctgtgg taccggcgtc agcgcctgct ggccaggacg ctgcggctgt ggttcgagaa
     1261 gagctcccgg ggccaggcga agctggagca gcggcggcag gaggaggccc tggagcgcaa
     1321 gaaggaggag gcgcggaggc ggcggcgcga ggtgaaacgg cggctgctgg aggagctggc
     1381 cagcacccct ccgcgcaaga gtgcacggct gctcctgcac acgccccgcg acgcgggccc
     1441 ggctgcctcg cgccccgccg ccctccctac ctcacgcaga gtgcccgccg ccacactcaa
     1501 gctgcgtcgc cagccggcca tggccccagg cgtcacgggc ctgcgcaacc tgggcaacac
     1561 ctgctacatg aactccatcc tccaggtgct cagccacctc cagaagttcc gagaatgttt
     1621 cctcaacctt gacccttcca aaacggaaca tctgtttccc aaagccacca acgggaagac
     1681 tcagctttct ggcaagccaa ccaacagctc ggccacggag ctgtccttga gaaatgacag
     1741 ggccgaggca tgcgagcggg agggcttctg ctggaacggc agggcctcca ttagtcggag
     1801 tctggagctc atccagaaca aggagccgag ttcaaagcac atttccctct gccgtgaact
     1861 gcacaccctc ttccgagtca tgtggtccgg gaagtgggcc ctagtgtcgc ccttcgccat
     1921 gctccactca gtgtggagcc tgatccctgc cttccgcggc tacgaccaac aggacgcgca
     1981 ggaatttctc tgcgagctgc tgcacaaggt gcagcaggaa ctcgagtctg agggcaccac
     2041 acgccggatc ctcatcccct tctcccagag gaagctcacc aaacaggtct taaaggtggt
     2101 gaataccata tttcatgggc agctgctcag tcaggtcaca tgtatatcat gcaattacaa
     2161 atccaatacc attgagccct tttgggacct atccctggaa ttccctgaac gctatcactg
     2221 catagaaaag gggtttgtcc ctttgaatca aacagagtgc ttgctcactg agatgctggc
     2281 caaattcaca gagacagagg ccctggaagg gagaatctac gcttgtgacc agtgtaacag
     2341 caaacgacga aaatccaatc ccaaacccct tgttctgagt gaagctagaa agcagttaat
     2401 gatctacaga ctacctcagg ttctccggct gcaccttaaa agattcaggt ggtctggccg
     2461 taatcatcga gagaagattg gggtccatgt cgtctttgac caggtattaa ccatggaacc
     2521 ttactgctgc agggacatgc tctcctctct tgacaaagag acctttgcct atgatctctc
     2581 cgcagtggtc atgcatcacg ggaaagggtt tggctcagga cactacacag cctattgcta
     2641 caacacagag ggaggttttt gggtccactg caatgactca aagctgaatg tatgcagtgt
     2701 cgaggaagtg tgcaaaaccc aggcctacat ccttttttac actcaaagaa cagtgcaggg
     2761 caatgcaaga atctcagaaa cccatctcca agctcaggtg cagtccagca acaatgatga
     2821 aggcagacca cagacatttt ccctcgagga ctacaaagac catgacggtg attataaaga
     2881 tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgata tctcgacaat
     2941 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     3001 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     3061 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     3121 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     3181 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     3241 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     3301 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc
     3361 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     3421 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     3481 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg aaacggggga
     3541 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     3601 aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     3661 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     3721 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     3781 gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag ggggtatccc
     3841 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3901 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3961 acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt
     4021 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     4081 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4141 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4201 taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4261 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4321 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4381 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4441 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4501 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     4561 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     4621 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     4681 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4741 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4801 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4861 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4921 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4981 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     5041 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     5101 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     5161 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     5221 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     5281 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     5341 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5401 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5461 acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac
     5521 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     5581 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     5641 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     5701 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     5761 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     5821 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5881 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5941 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     6001 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     6061 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     6121 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     6181 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6241 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6301 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6361 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
     6421 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6481 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6541 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6601 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6661 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6721 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6781 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6841 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6901 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6961 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     7021 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     7081 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7141 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7201 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     7261 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     7321 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7381 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7441 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7501 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7561 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     7621 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7681 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7741 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7801 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7861 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7921 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     7981 ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc tctgatgccg
     8041 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     8101 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     8161 gg
//
